https://wiki.houptlab.org/api.php?action=feedcontributions&feedformat=atom&user=HouptMagnetoWiki - User contributions [en]2026-04-09T14:30:33ZUser contributionsMediaWiki 1.35.8https://wiki.houptlab.org/index.php?title=SNB_Lithium_Orotate_CTA_2&diff=3334SNB Lithium Orotate CTA 22026-03-20T15:12:20Z<p>Houpt: added pH details</p>
<hr />
<div><br />
<br />
Continuation of [[SNA Lithium Orotate CTA]].<br />
<br />
Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl, with a lower dose than SNA experiment<br />
<br />
Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt2<br />
| project = Taste Aversion<br />
| code = SNB<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2026-3-13<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNB<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNB Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-2-10), n= 16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2025-12-15), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-3-13)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-3-14 thru 2026-3-19)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 900 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-3-20)<br />
<br />
0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
<br />
30 min: rats injected with:<br />
<br />
* 75 mM NaCl, 6 ml/kg ip. (SNB01,04,07,10,13)<br />
* 75 mM LiCl, 6 ml/kg ip. (SNB02,05,08,11,14)<br />
* 75 mM LiOr, 6 ml/kg ip. (SNB03,06,09,12,15,16)<br />
<br />
all injections made isotonic with TrisAmino:<br />
<br />
* 0.3 M TrisAmino <br />
36.34g / liter, autoclave, pH to 7.4 with 38% HCl<br />
<br />
* 0.15 M LiOH<br />
3.6g / liter (no need to autoclave)<br />
<br />
* 0.15 M LiCl<br />
6.45g / liter, autoclave<br />
<br />
* LiOr = 75mM LiOrotate, 150mM TrisAmino<br />
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiOH, 0.47g orotic acid, pH to 7.4 with LiOH, filter<br />
<br />
* LiCl = 75mM LiCl, 150mM TrisAmino<br />
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiCl, pH to 7.4 with LiOH, filter<br />
<br />
* NaCl = 75mM NaCl, 150 mM TrisAmino<br />
20 ml of 0.3M TrisAmino, 20 ml of 0.15M NaCl, pH to 7.4 with NaOH, filter<br />
<br />
<br />
Ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-3-21 through 2026-4-1)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=SNB_Lithium_Orotate_CTA_2&diff=3333SNB Lithium Orotate CTA 22026-03-18T15:36:48Z<p>Houpt: added Li and Na injection prep</p>
<hr />
<div><br />
<br />
Continuation of [[SNA Lithium Orotate CTA]].<br />
<br />
Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl, with a lower dose than SNA experiment<br />
<br />
Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt2<br />
| project = Taste Aversion<br />
| code = SNB<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2026-3-13<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNB<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNB Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-2-10), n= 16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2025-12-15), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-3-13)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-3-14 thru 2026-3-19)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 900 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-3-20)<br />
<br />
0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
<br />
30 min: rats injected with:<br />
<br />
* 75 mM NaCl, 6 ml/kg ip. (SNB01,04,07,10,13)<br />
* 75 mM LiCl, 6 ml/kg ip. (SNB02,05,08,11,14)<br />
* 75 mM LiOr, 6 ml/kg ip. (SNB03,06,09,12,15,16)<br />
<br />
all injections made isotonic with TrisAmino:<br />
<br />
* 0.3 M TrisAmino <br />
36.34g / liter<br />
<br />
* 0.15 M LiOH<br />
3.6g / liter<br />
<br />
* 0.15 M LiCl<br />
6.45g / liter<br />
<br />
* LiOr = 75mM LiOrotate, 150mM TrisAmino<br />
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiOH, 0.47g orotic acid<br />
<br />
* LiCl = 75mM LiCl, 150mM TrisAmino<br />
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiCl<br />
<br />
* NaCl = 75mM NaCl, 150 mM TrisAmino<br />
20 ml of 0.3M TrisAmino, 20 ml of 0.15M NaCl<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-3-21 through 2026-4-1)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3332Lithium Salts2026-03-18T15:35:13Z<p>Houpt: added LiOH</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
NaOr 178.08 (0.13 Na) 26.71 12 320.54 (41.4 Na)<br />
LiOH 23.95 0.29 3.6 na na na<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4. Doses were administered as 0.1 ml/10g BW (10 ml/kg), with the highest dose of 15 mg/kg Li+ => 375 mg/kg LiOr => 230 mM (NB: not sure how they got this much to dissolve in water)<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1, MW 156.10<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2, MW 23.95<br />
<br />
sodium hydroxide, CAS 1310-73-2, MW 40<br />
<br />
* Make 500 ml of 0.15M LiOH (1.80 g LiOH / 500ml dH<small>2</small>0).<br />
<br />
* Make 500 ml of 0.15M NaOH (3.00 g NaOH / 500ml dH<small>2</small>0).<br />
<br />
* Make 20 ml of 0.15M LiOr by adding 0.47 g orotic acid to 20 ml of 0.15M LiOH, vortex: partially dissolved after about 2 min of vortexing -- but soon precipiates out<br />
<br />
* So dissolve in 0.3M TrisAmino...<br />
<br />
* Make 20 ml of 0.15M NaOr by adding 0.47 g orotic acid to 20 ml of 0.15M NaOH, vortex: did NOT dissolve -- milky white suspension! Left shaking overnight...<br />
<br />
<br />
Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides<br />
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So may need to mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=SNB_Lithium_Orotate_CTA_2&diff=3331SNB Lithium Orotate CTA 22026-03-14T15:44:36Z<p>Houpt: updated dates</p>
<hr />
<div><br />
<br />
Continuation of [[SNA Lithium Orotate CTA]].<br />
<br />
Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl, with a lower dose than SNA experiment<br />
<br />
Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt2<br />
| project = Taste Aversion<br />
| code = SNB<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2026-3-13<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNB<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNB Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-2-10), n= 16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2025-12-15), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-3-13)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-3-14 thru 2026-3-19)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 900 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-3-20)<br />
<br />
0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
<br />
30 min: rats injected with:<br />
<br />
* 75 mM NaCl, 6 ml/kg ip. (SNB01,04,07,10,13)<br />
* 75 mM LiCl, 6 ml/kg ip. (SNB02,05,08,11,14)<br />
* 75 mM LiOr, 6 ml/kg ip. (SNB03,06,09,12,15,16)<br />
<br />
all injections made isotonic with TrisAmino<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-3-21 through 2026-4-1)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=SNA_Lithium_Orotate_CTA&diff=3330SNA Lithium Orotate CTA2026-03-14T14:01:17Z<p>Houpt: /* Agenda */Added dob</p>
<hr />
<div>Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl. <br />
<br />
Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt1<br />
| project = Taste Aversion<br />
| code = SNA<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2026-1-14<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNA<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNA Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-1-20), n= 16 adult male Sprague Dawley rats d.o.b. 2025-11-24 (Charles River CD), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-1-26)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-1-27 thru 2026-2-1)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-2-2)<br />
<br />
0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
<br />
30 min: rats injected with:<br />
<br />
* 0.15 M NaCl, 6 ml/kg ip. (SNA01,04,07,10,13)<br />
* 0.15 M LiCl, 6 ml/kg ip. (SNA02,05,08,11,14)<br />
* 0.15 M LiOr, 6 ml/kg ip. (SNA03,06,09,12,15,16) (LiOr was more of a milky suspension)<br />
<br />
LiOr prepared as orotic acid added to 0.15M LiOH to make nominal 0.15M Li Orotate -- but did not go competely into solution, so injected "slurry"<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 34: Preference Tests''' (2026-2-3 through 2026-2-24)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 21 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=SNA_Lithium_Orotate_CTA&diff=3329SNA Lithium Orotate CTA2026-03-12T12:54:31Z<p>Houpt: updated some dates, LiOr prep</p>
<hr />
<div>Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl. <br />
<br />
Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt1<br />
| project = Taste Aversion<br />
| code = SNA<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2026-1-14<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNA<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNA Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-1-20), n= 16 adult male Sprague Dawley rats d.o.b. 2025-????? (Charles River CD), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-1-26)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-1-27 thru 2026-2-1)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-2-2)<br />
<br />
0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
<br />
30 min: rats injected with:<br />
<br />
* 0.15 M NaCl, 6 ml/kg ip. (SNA01,04,07,10,13)<br />
* 0.15 M LiCl, 6 ml/kg ip. (SNA02,05,08,11,14)<br />
* 0.15 M LiOr, 6 ml/kg ip. (SNA03,06,09,12,15,16) (LiOr was more of a milky suspension)<br />
<br />
LiOr prepared as orotic acid added to 0.15M LiOH to make nominal 0.15M Li Orotate -- but did not go competely into solution, so injected "slurry"<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 34: Preference Tests''' (2026-2-3 through 2026-2-24)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 21 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=SNB_Lithium_Orotate_CTA_2&diff=3328SNB Lithium Orotate CTA 22026-03-12T12:49:59Z<p>Houpt: added link to prior experiment</p>
<hr />
<div><br />
<br />
Continuation of [[SNA Lithium Orotate CTA]].<br />
<br />
Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl, with a lower dose than SNA experiment<br />
<br />
Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt2<br />
| project = Taste Aversion<br />
| code = SNB<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2026-3-13<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNB<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNB Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-2-10), n= 16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2025-12-15), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-3-12)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-3-13 thru 2026-3-18)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 900 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-3-19)<br />
<br />
0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
<br />
30 min: rats injected with:<br />
<br />
* 75 mM NaCl, 6 ml/kg ip. (SNB01,04,07,10,13)<br />
* 75 mM LiCl, 6 ml/kg ip. (SNB02,05,08,11,14)<br />
* 75 mM LiOr, 6 ml/kg ip. (SNB03,06,09,12,15,16)<br />
<br />
all injections made isotonic with TrisAmino<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-3-20 through 2026-3-31)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=SNB_Lithium_Orotate_CTA_2&diff=3327SNB Lithium Orotate CTA 22026-03-12T12:48:49Z<p>Houpt: created page</p>
<hr />
<div>Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl. <br />
<br />
Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt2<br />
| project = Taste Aversion<br />
| code = SNB<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2026-3-13<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNB<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNB Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-2-10), n= 16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2025-12-15), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-3-12)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-3-13 thru 2026-3-18)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 900 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-3-19)<br />
<br />
0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
<br />
30 min: rats injected with:<br />
<br />
* 75 mM NaCl, 6 ml/kg ip. (SNB01,04,07,10,13)<br />
* 75 mM LiCl, 6 ml/kg ip. (SNB02,05,08,11,14)<br />
* 75 mM LiOr, 6 ml/kg ip. (SNB03,06,09,12,15,16)<br />
<br />
all injections made isotonic with TrisAmino<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-3-20 through 2026-3-31)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=MNase_Digestion&diff=3326MNase Digestion2026-02-04T19:33:10Z<p>Houpt: typo</p>
<hr />
<div>* injecting the rats with sodium pentobarbitol<br />
* transcardial perfusion 100ml/2min Saline nitride heparine for each rat<br />
* transcardial perfusion with 400ml/4min 1% phospahte buffered paraformaldehyde<br />
* Dissect rat [[Adrenal Gland]] and put in PBS on ice and tranfer to the lab <br />
* Separate medulla in PBS (the tissues were in PBS around 60 min)<br />
* 4 medullas (~4 x 10<sup>6</sup> cells total) in Dounce homogenizer in 1ml PBS, <br />
* Quenching by adding with 1/20 of stock Glycine (2.5M), final concentration is 125mM<br />
* homogenize the tissues and let the tube sit for 5 min at RT<br />
* Spinning down the cells at 16000rcf, supernatant is removed<br />
* The cell pellet is resuspended in 1ml Nucleus Isolation Buffer (NIB) (10mM HEPES at pH 7.8, 2mM MgoAc2, 0.3M sucrose, 2mM CaCl2, 1%Triton-X) let the tube sit for 10 min at RT<br />
* Spinning down the cells at 1000rcf, supernatant is removed<br />
* resuspend the cells in 1ml NIB <br />
* Spinning down the cells at 1000rcf, supernatant is removed<br />
* resuspending the cells in 500 µl in NIB<br />
* prewarm the cells ar 37°C form 1 min<br />
* Prepare 2 MNase digestion reaction (total volume 250µl each), MNase stock concentration : 20U/µl diluted to 2 U/µl<br />
* 1: add 5µl of 2U/µl to get 10U, 10min, RT<br />
* 2: add 2.5µl of 20U/µl to get 50U, 10min, RT<br />
* Stop the reaction with 10 µl EDTA(500mM), the final concentration was 20mM<br />
* Add 6.25µl of 20%SDS (final concentration 0.5%)<br />
* Adding 2.5µl of Proteinase K 20mg/µl) (final concentration 0.2 µg/l)<br />
* Incubate at first 55 C for 1-2 hours and then overnight at 65 C<br />
* add 2.4µl RNAse (final concentration 50µg/ml) for 20min at 37° C<br />
* DNA Purification with just Phenol:cholorophorm:isoamyl and Ethanol precipitation</div>Houpthttps://wiki.houptlab.org/index.php?title=MNase_Digestion&diff=3325MNase Digestion2026-02-04T19:31:29Z<p>Houpt: added adrenal link and number of cells</p>
<hr />
<div>* injecting the rats with sodium pentobarbitol<br />
* transcardial perfusion 100ml/2min Saline nitride heparine for each rat<br />
* transcardial perfusion with 400ml/4min 1% phospahte buffered paraformaldehyde<br />
* Dissect [[Arenal_Gland rat adrenal gland]] and put in PBS on ice and tranfer to the lab <br />
* Separate medulla in PBS (the tissues were in PBS around 60 min)<br />
* 4 medullas (~4 x 10<sup>6</sup> cells total) in Dounce homogenizer in 1ml PBS, <br />
* Quenching by adding with 1/20 of stock Glycine (2.5M), final concentration is 125mM<br />
* homogenize the tissues and let the tube sit for 5 min at RT<br />
* Spinning down the cells at 16000rcf, supernatant is removed<br />
* The cell pellet is resuspended in 1ml Nucleus Isolation Buffer (NIB) (10mM HEPES at pH 7.8, 2mM MgoAc2, 0.3M sucrose, 2mM CaCl2, 1%Triton-X) let the tube sit for 10 min at RT<br />
* Spinning down the cells at 1000rcf, supernatant is removed<br />
* resuspend the cells in 1ml NIB <br />
* Spinning down the cells at 1000rcf, supernatant is removed<br />
* resuspending the cells in 500 µl in NIB<br />
* prewarm the cells ar 37°C form 1 min<br />
* Prepare 2 MNase digestion reaction (total volume 250µl each), MNase stock concentration : 20U/µl diluted to 2 U/µl<br />
* 1: add 5µl of 2U/µl to get 10U, 10min, RT<br />
* 2: add 2.5µl of 20U/µl to get 50U, 10min, RT<br />
* Stop the reaction with 10 µl EDTA(500mM), the final concentration was 20mM<br />
* Add 6.25µl of 20%SDS (final concentration 0.5%)<br />
* Adding 2.5µl of Proteinase K 20mg/µl) (final concentration 0.2 µg/l)<br />
* Incubate at first 55 C for 1-2 hours and then overnight at 65 C<br />
* add 2.4µl RNAse (final concentration 50µg/ml) for 20min at 37° C<br />
* DNA Purification with just Phenol:cholorophorm:isoamyl and Ethanol precipitation</div>Houpthttps://wiki.houptlab.org/index.php?title=Adrenal_Gland&diff=3324Adrenal Gland2026-02-04T19:29:38Z<p>Houpt: calculalted number of cells</p>
<hr />
<div><br />
Some papers on [https://houptlab.org/courses/adrenal adrenal gland and regulation of the medulla].<br />
<br />
=Rat Adrenal Gland Facts=<br />
<br />
''From Tomlinson 1987 PMID 3601067''<br />
<br />
Adrenal Gland: 13.2 mm<sup>3</sup><br />
<br />
Adrenal Cortex: '''(NOTE: need to add number of cells)'''<br />
<br />
Adrenal Medulla: 1.3 mm<sup>3</sup><br />
<br />
Medulla by volume: 63% chromaffin (catecholamine) cells, 20% vasculature, 5% neural, 12% interstitial<br />
<br />
Medulla number of cells (unilateral):<br />
<br />
* 11.1 x10<sup>5</sup> total cells <br />
<br />
* 5 x10<sup>5</sup> epinephrine cells (TH & PNMT)<br />
<br />
* 2 x 10<sup>5</sup> norepinephrine cells (TH)</div>Houpthttps://wiki.houptlab.org/index.php?title=SNA_Lithium_Orotate_CTA&diff=3323SNA Lithium Orotate CTA2026-02-03T14:56:06Z<p>Houpt: updated groups</p>
<hr />
<div>Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl. <br />
<br />
Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt1<br />
| project = Taste Aversion<br />
| code = SNA<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2025-1-14<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNA<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNA Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-1-20), n= 16 adult male Sprague Dawley rats (Charles River CD), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-1-26)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-1-27 thru 2026-2-1)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-2-2)<br />
<br />
0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
<br />
30 min: rats injected with:<br />
<br />
* 0.15 M NaCl, 6 ml/kg ip. (SNA01,04,07,10,13)<br />
* 0.15 M LiCl, 6 ml/kg ip. (SNA02,05,08,11,14)<br />
* 0.15 M LiOr, 6 ml/kg ip. (SNA03,06,09,12,15,16) (LiOr was more of a milky suspension)<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-2-3 through 2026-2-13)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3322Lithium Salts2026-01-30T21:17:49Z<p>Houpt: </p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
NaOr 178.08 (0.13 Na) 26.71 12 320.54 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4. Doses were administered as 0.1 ml/10g BW (10 ml/kg), with the highest dose of 15 mg/kg Li+ => 375 mg/kg LiOr => 230 mM (NB: not sure how they got this much to dissolve in water)<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1, MW 156.10<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2, MW 23.95<br />
<br />
sodium hydroxide, CAS 1310-73-2, MW 40<br />
<br />
* Make 500 ml of 0.15M LiOH (1.80 g LiOH / 500ml dH<small>2</small>0).<br />
<br />
* Make 500 ml of 0.15M NaOH (3.00 g NaOH / 500ml dH<small>2</small>0).<br />
<br />
* Make 20 ml of 0.15M LiOr by adding 0.47 g orotic acid to 20 ml of 0.15M LiOH, vortex: dissolved after about 2 min of vortexing, store at 4C in foil-wrapped tube.<br />
<br />
* Make 20 ml of 0.15M NaOr by adding 0.47 g orotic acid to 20 ml of 0.15M NaOH, vortex: did NOT dissolve -- milky white suspension! Left shaking overnight...<br />
<br />
<br />
Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides<br />
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So may need to mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3321Lithium Salts2026-01-29T16:31:33Z<p>Houpt: revised LiOr preparation</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
NaOr 178.08 (0.13 Na) 26.71 12 320.54 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4. Doses were administered as 0.1 ml/10g BW (10 ml/kg), with the highest dose of 15 mg/kg Li+ => 375 mg/kg LiOr => 230 mM (NB: not sure how they got this much to dissolve in water)<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1, MW 156.10<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2, MW 23.95<br />
<br />
sodium hydroxide, CAS 1310-73-2, MW 40<br />
<br />
* Make 500 ml of 0.15M LiOH (1.80 g LiOH / 500ml dH<small>2</small>0).<br />
<br />
* Make 500 ml of 0.15M NaOH (3.00 g NaOH / 500ml dH<small>2</small>0).<br />
<br />
* Make 20 ml of 0.15M LiOr by adding 0.47 g orotic acid to 20 ml of 0.15M LiOH (need to heat?)<br />
<br />
* Make 20 ml of 0.15M NaOr by adding 0.47 g orotic acid to 20 ml of 0.15M NaOH (need to heat?)<br />
<br />
<br />
Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides<br />
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So may need to mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3320Lithium Salts2026-01-27T17:27:29Z<p>Houpt: typo</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
NaOr 178.08 (0.13 Na) 26.71 12 320.54 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4.<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1, MW 156.10<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2, MW 23.95<br />
<br />
0.15M LiOr: mix equal volumes of 0.3 M LiOH and 0.3 M orotic acid<br />
<br />
0.15M NaOr: mix equal volumes of 0.3 M NaOH and 0.3 M orotic acid<br />
<br />
* 0.3M orotic acid (MW 156.10) = 46.83 g / liter, 0.94 g / 20 ml<br />
* 0.3M LiOH (MW 23.95) = 7.19 g / liter, 0.14 g / 20 ml<br />
* 0.3M NaOH (MW 39.997) = 12 g / liter, 0.24 g/20ml<br />
<br />
<br />
<br />
Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides<br />
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So we'll mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3319Lithium Salts2026-01-27T17:25:40Z<p>Houpt: added 0.15M LiOr preparation</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
NaOr 178.08 (0.13 Na) 26.71 12 320.54 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4.<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1, MW 46.83<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2, MW 23.95<br />
<br />
0.15M LiOr: mix equal volumes of 0.3 M LiOH and 0.3 M orotic acid<br />
<br />
0.15M NaOr: mix equal volumes of 0.3 M NaOH and 0.3 M orotic acid<br />
<br />
* 0.3M orotic acid (MW 46.83) = 46.83 g / liter, 0.94 g / 20 ml<br />
* 0.3M LiOH (MW 23.95) = 7.19 g / liter, 0.14 g / 20 ml<br />
* 0.3M NaOH (MW 39.997) = 12 g / liter, 0.24 g/20ml<br />
<br />
<br />
<br />
Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides<br />
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So we'll mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=SNA_Lithium_Orotate_CTA&diff=3318SNA Lithium Orotate CTA2026-01-14T20:25:26Z<p>Houpt: </p>
<hr />
<div>Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl. <br />
<br />
Pair saccharin intake with different doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt1<br />
| project = Taste Aversion<br />
| code = SNA<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2025-1-14<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNA<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNA Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-1-20), n= 16 adult male Sprague Dawley rats (Charles River CD), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-1-26)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-1-27 thru 2026-2-1)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-2-2)<br />
<br />
0 min: 30 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
30 min: rats injected with:<br />
<br />
* 0.15 M NaCl, 6 ml/kg ip.<br />
* 0.15 M LiCl, 6 ml/kg ip.<br />
* 0.15 M NaOr, 6 ml/kg ip.<br />
* 0.15 M LiOr, 6 ml/kg ip.<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-2-3 through 2026-2-13)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=SNA_Lithium_Orotate_CTA&diff=3317SNA Lithium Orotate CTA2026-01-14T20:21:50Z<p>Houpt: added labels</p>
<hr />
<div>Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl. <br />
<br />
Pair saccharin intake with different doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt1<br />
| project = Taste Aversion<br />
| code = SNA<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2025-1-14<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNA<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNA Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-1-20), n= 16 adult male Sprague Dawley rats (Charles River CD), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-1-26)<br />
<br />
make labels for bottles.<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-1-27 thru 2026-2-1)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-2-2)<br />
<br />
0 min: 30 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
30 min: rats injected with:<br />
<br />
* 0.15 M NaCl, 6 ml/kg ip.<br />
* 0.15 M LiCl, 6 ml/kg ip.<br />
* 0.15 M NaOr, 6 ml/kg ip.<br />
* 0.15 M LiOr, 6 ml/kg ip.<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-2-3 through 2026-2-13)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]<br />
[[Category:Epigenetics]]</div>Houpthttps://wiki.houptlab.org/index.php?title=SNA_Lithium_Orotate_CTA&diff=3316SNA Lithium Orotate CTA2026-01-14T20:13:44Z<p>Houpt: typo</p>
<hr />
<div>Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl. <br />
<br />
Pair saccharin intake with different doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt1<br />
| project = Taste Aversion<br />
| code = SNA<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2025-1-14<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNA<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNA Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-1-20), n= 16 adult male Sprague Dawley rats (Charles River CD), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-1-26)<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-1-27 thru 2026-2-1)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-2-2)<br />
<br />
0 min: 30 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
30 min: rats injected with:<br />
<br />
* 0.15 M NaCl, 6 ml/kg ip.<br />
* 0.15 M LiCl, 6 ml/kg ip.<br />
* 0.15 M NaOr, 6 ml/kg ip.<br />
* 0.15 M LiOr, 6 ml/kg ip.<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-2-3 through 2026-2-13)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]<br />
[[Category:Epigenetics]]</div>Houpthttps://wiki.houptlab.org/index.php?title=SNA_Lithium_Orotate_CTA&diff=3315SNA Lithium Orotate CTA2026-01-14T20:13:12Z<p>Houpt: first draft of experiment</p>
<hr />
<div>Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl. <br />
<br />
Pair saccharin intake with different doses of LiCl and LiOr to see if LiOr is more potent.<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Lithium Orotate and CTA pt1<br />
| project = Taste Aversion<br />
| code = SNA<br />
| protocol = 20220000006<br />
| funding = Chair's Fund<br />
| investigators = Nabi, Houpt<br />
| species = rat<br />
| date = 2025-1-14<br />
| type = CTA<br />
| drugs = lithium orotate, lithium chloride<br />
| data = https://houptlab.org/bartab/expt/?id=SNA<br />
}}<br />
<br />
== Data ==<br />
<br />
[https://houptlab.org/bartab/expt/?id=SNA Raw Data Link]<br />
<br />
== People ==<br />
<br />
Soheila Nabi<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
Rats arrive (2026-1-20), n= 16 adult male Sprague Dawley rats (Charles River CD), individually housed in BRF 169.<br />
<br />
<br><br />
<br />
'''Day 0: Water removed''' (2026-1-26)<br />
<br />
<br><br />
<br />
'''Days 1 - 6: Water Restriction''' (2026-1-27 thru 2026-2-1)<br />
<br />
Weigh rats daily. ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily.<br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 12: Conditioning Day''' (2026-2-2)<br />
<br />
0 min: 30 min access to 0.125% saccharin (5 g sodium saccharin/4L water)<br />
30 min: rats injected with:<br />
<br />
- 0.15 M NaCl, 6 ml/kg ip.<br />
- 0.15 M LiCl, 6 ml/kg ip.<br />
- 0.15 M NaOr, 6 ml/kg ip.<br />
- 0.15 M LiOr, 6 ml/kg ip.<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
<br />
'''Days 13 - 22: Preference Tests''' (2026-2-3 through 2026-2-13)<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]<br />
[[Category:Epigenetics]]</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3314Lithium Salts2026-01-05T16:24:38Z<p>Houpt: added ordering info for orotic acid and LiOH</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
NaOr 178.08 (0.13 Na) 26.71 12 320.54 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4.<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1<br />
<br />
[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2<br />
<br />
<br />
Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides<br />
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So we'll mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=File:RatNORArenaSchematic.png&diff=3312File:RatNORArenaSchematic.png2025-11-14T19:36:21Z<p>Houpt: Schematic of arena for rat novel object recognition test</p>
<hr />
<div>== Summary ==<br />
Schematic of arena for rat novel object recognition test</div>Houpthttps://wiki.houptlab.org/index.php?title=Novel_Object_Recognition&diff=3311Novel Object Recognition2025-11-14T19:33:40Z<p>Houpt: added kongs photo</p>
<hr />
<div><br />
==Rat Setup==<br />
<br />
===Arena===<br />
<br />
external size: 79.2 cm square (not including handles), with 112cm diagonal -- so will fit on a 48" round table.<br />
<br />
interior size: 70cm square, with 35cm high walls.<br />
<br />
<br />
<br />
Place 36x36" sterilization wrap from Schein for substrate. [https://www.henryschein.com/us-en/dental/p/infection-control-products/instrument-sterilization-processing/csr-wrap/1002016 CSR Wrap 36x36" 100/Bx 1002016EZ | Henry Schein Inc. - 849]<br />
<br />
Constructed from [8020.net 8020] materials.<br />
<br />
===Objects===<br />
<br />
Kong Classic Dog Toy (large) vs. Kong Jump'N Jack Dog Toy (medium).<br />
<br />
''Note that Jump'N Jack has been discontinued.''<br />
<br />
[[File:Kongs4NOR.jpg|200px|thumb|left|Kongs used for rat NOR test]]</div>Houpthttps://wiki.houptlab.org/index.php?title=File:Kongs4NOR.jpg&diff=3310File:Kongs4NOR.jpg2025-11-14T19:32:19Z<p>Houpt: Photos of Kong Classic and Kong Jump'N Jack Dog Toys used for rat novel object recognition test.</p>
<hr />
<div>== Summary ==<br />
Photos of Kong Classic and Kong Jump'N Jack Dog Toys used for rat novel object recognition test.</div>Houpthttps://wiki.houptlab.org/index.php?title=Novel_Object_Recognition&diff=3309Novel Object Recognition2025-11-14T19:31:02Z<p>Houpt: create page</p>
<hr />
<div><br />
==Rat Setup==<br />
<br />
===Arena===<br />
<br />
external size: 79.2 cm square (not including handles), with 112cm diagonal -- so will fit on a 48" round table.<br />
<br />
interior size: 70cm square, with 35cm high walls.<br />
<br />
<br />
<br />
Place 36x36" sterilization wrap from Schein for substrate. [https://www.henryschein.com/us-en/dental/p/infection-control-products/instrument-sterilization-processing/csr-wrap/1002016 CSR Wrap 36x36" 100/Bx 1002016EZ | Henry Schein Inc. - 849]<br />
<br />
Constructed from [8020.net 8020] materials.<br />
<br />
===Objects===<br />
<br />
Kong Classic Dog Toy (large) vs. Kong Jump'N Jack Dog Toy (medium).<br />
<br />
''Note that Jump'N Jack has been discontinued.''</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3308Lithium Salts2025-10-09T13:58:30Z<p>Houpt: added sodium orotate</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
NaOr 178.08 (0.13 Na) 26.71 12 320.54 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was<br />
adjusted to 0.9% and pH to 7.4.<br />
<br />
Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides<br />
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So we'll mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=Pica&diff=3307Pica2025-09-29T16:43:20Z<p>Houpt: added Physics-Integrated Clothed Avatar</p>
<hr />
<div>Multiple uses of the word Pica or Pika<br />
<br />
<br />
=Pika= <br />
<br />
rock rabbit<br />
<br />
a lagomorph (relative of rabbits) in the mountains of the western United States<br />
<br />
=Pica= <br />
<br />
eating clay or other nonnutirive subantances to detoxify ingested toxins. Named after the behavior of magpies.<br />
<br />
Can be [https://newsfeed.time.com/2013/01/30/japanese-restaurant-serves-meal-of-dirt-for-110/ pricey.]<br />
<br />
=Pica Pica=<br />
<br />
The European Magpie <br />
<br />
=Hyrax=<br />
<br />
[https://en.wikipedia.org/wiki/Hyrax Not] a pica. Also not a rodent; related to elephants.<br />
<br />
=Puca=<br />
<br />
A creature of Celtic folklore, Though the púca enjoys confusing and often terrifying humans, it is considered to be benevolent.<br />
<br />
=Pooka=<br />
<br />
https://en.wikipedia.org/wiki/Harvey_(film)<br />
<br />
=Pica=<br />
<br />
A typographic unit of measure corresponding to 1/72nd of its respective foot, and therefore to 1/6th of an inch. .<br />
<br />
=Pica, Chile=<br />
<br />
[https://en.wikipedia.org/wiki/Pica,_Chile Pica] in the north of Chile is famous for its "small and unusually acidic lemons." Coincidentally, it is also the site of substantial [https://www.mining.com/mammoth-energy-group-continues-to-acquire-chilean-lithium-assets/ lithium concessions] acquired by Mammoth Energy Group's Compania Lithium Investments Limitada, from the dry lake bed of Laguna Lagunilla.<br />
<br />
=Pika=<br />
<br />
[https://www.pika.dev/about Pika] is a “project to move the JavaScript ecosystem forward.” Allows searching for ES module syntax packages, and integrates ESM packages with npm.<br />
<br />
Has an icon of a pika for a mascot.<br />
<br />
=Piku=<br />
<br />
How I love a verse<br />
Contrived to unhusk dryly<br />
One image nutshell<br />
<br />
(count the letters)<br />
<br />
=Pika Pika=<br />
<br />
[https://nakamotoforestry.com/shou-sugi-ban-siding/ Pika Pika] ( ぴかぴか ) is variant of charred cedar boards (焼杉板 or “yakisugi-ita”) used for siding in Japanese building. The charing helps preserve the boards. Pika Pika boards are scrapped with a wire brush that increases contrast by removing soot from softer grain in the wood. <br />
<br />
=PikaPython=<br />
<br />
[https://github.com/pikasTech/PikaPython PikaPython] is a cross platform ultra lightweight embedded Python engine.<br />
<br />
=PICA=<br />
<br />
Paillier-based InChain Aggregation PMID 39275535<br />
<br />
=PICA=<br />
<br />
Physics-Integrated Clothed Avatar PMID 41004371<br />
[[Category:Taste Aversion]]</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3306Lithium Salts2025-09-16T14:44:05Z<p>Houpt: added naringin and azauracil doses</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was<br />
adjusted to 0.9% and pH to 7.4.<br />
<br />
Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides<br />
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So we'll mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3305Lithium Salts2025-09-15T19:54:27Z<p>Houpt: fixed LiOr concentration bug</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 24.31 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was<br />
adjusted to 0.9% and pH to 7.4.<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So we'll mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3304Lithium Salts2025-09-13T18:11:26Z<p>Houpt: added LiOr prep</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 01.04 12 291.79 12.5<br />
</tab><br />
<br />
== LiOr Preparation ==<br />
<br />
From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was<br />
adjusted to 0.9% and pH to 7.4.<br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So we'll mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=Lithium_Salts&diff=3303Lithium Salts2025-09-13T17:59:28Z<p>Houpt: created page for orotate experiment</p>
<hr />
<div><br />
MW of lithium: 6.94 (weighted average of Li<sup>6</sup> and Li<sup>7</sup> isotopes)<br />
<br />
For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day). <br />
<br />
Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl. <br />
<br />
We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li<sub>2</sub>CO<sub>3</sub>; LCO) and lithium orotate (LiH<sub>3</sub>N<sub>2</sub>O<sub>4</sub>; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:<br />
<br />
<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top><br />
Salt MW % Li g/L for 0.15M dose ml 0.15M/kg mg salt/kg mg Li/kg<br />
NaCl 58.44 (0.39 Na) 8.8 12 105.19 (41.4 Na)<br />
LiCl 42.39 0.16 6.35 12 76.30 12.5<br />
LiCl 42.39 0.16 6.35 20 127.17 41.6<br />
LiCO 73.89 0.19 11.08 12 66.54 12.5<br />
LiOr 162.00 0.04 01.04 12 291.79 12.5<br />
</tab><br />
<br />
== Milliequivalents ==<br />
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.<br />
<br />
== Osmolarity ==<br />
<br />
NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).<br />
<br />
LiCO should also dissociate completely (citation needed).<br />
<br />
LiOr, however is more tightly bound complex (citation needed). So we'll mix up LiOr in 0.15 NaCl to preserve osmolarity.</div>Houpthttps://wiki.houptlab.org/index.php?title=DNA_Facts&diff=3301DNA Facts2025-07-09T17:42:28Z<p>Houpt: fixed TSS calculation</p>
<hr />
<div><br />
Some useful facts about DNA (taken from Wikipedia unless otherwise noted)<br />
<br />
mass of DNA: 1 bp 660 Daltons (660 g/mol)<br />
<br />
molarity of 1 ug/ul of single base-pairs = 1.5 mM (divide by fragment length to get molarity of fragments)<br />
<br />
DNA per mammalian cell: 6 pg<br />
<br />
length of DNA (diploid human) per cell: 2 m<br />
<br />
==Human Genome==<br />
<br />
human genome: 3.1 Gb, so 6 billion bp in normal diploid<br />
<br />
human chromosomes: 23 pairs (22 automosomes and X and Y chromosomes) <br />
<br />
human mitochondria genome: 16.5 kbp, encodes 13 proteins; 1% of cellular DNA (1000 - 10,000 copies per cell)<br />
<br />
<br />
==Rodent Genomes==<br />
<br />
rat genome: 2.87 Gb<br />
<br />
rat chromosomes: 21 pairs<br />
<br />
mouse genome: 2.73 Gb<br />
<br />
mouse chromosomes: 20 pairs<br />
<br />
==Nucleosomes==<br />
<br />
Approximately 30 million nucleosomes per human cell (~ 1 / 200 bp)<br />
<br />
147 bp wrapped (1.67 left-handed superhelical turns) around histone octamer (2 copies each H2A, H2B, H3, and H4). <br />
<br />
The nucleosome contains over 120 direct protein-DNA interactions and several hundred water-mediated ones.<br />
<br />
Linker DNA between nucleosomes 10-80 bp<br />
<br />
ideal MNase mononucleosomal digest -> 3 x 10<sup>7</sup> fragments per cell (3 x 10<sup>13</sup> for 1 million cells); if 4 x 10<sup>5</sup> transcription start sites (TSS) of 2000 bp length, so ~10 nucleosomes/TSS, then ~ 4 x10<sup>6</sup> (~10%) of mononucleosomal fragments will be of interest.<br />
<br />
Acetylation sites: "On average, 5 to 9 lysine residues of H2B, H3, and H4 were acetylated while a higher number (10 to 11 sites) of modifications was observed for H2A [but may be more]" PMID 37011222 ; so total of 50 - 76 acetylation sites per nucleosome.</div>Houpthttps://wiki.houptlab.org/index.php?title=DNA_Facts&diff=3300DNA Facts2025-07-09T17:20:37Z<p>Houpt: added some nucleosome and acetylation counts</p>
<hr />
<div><br />
Some useful facts about DNA (taken from Wikipedia unless otherwise noted)<br />
<br />
mass of DNA: 1 bp 660 Daltons (660 g/mol)<br />
<br />
molarity of 1 ug/ul of single base-pairs = 1.5 mM (divide by fragment length to get molarity of fragments)<br />
<br />
DNA per mammalian cell: 6 pg<br />
<br />
length of DNA (diploid human) per cell: 2 m<br />
<br />
==Human Genome==<br />
<br />
human genome: 3.1 Gb, so 6 billion bp in normal diploid<br />
<br />
human chromosomes: 23 pairs (22 automosomes and X and Y chromosomes) <br />
<br />
human mitochondria genome: 16.5 kbp, encodes 13 proteins; 1% of cellular DNA (1000 - 10,000 copies per cell)<br />
<br />
<br />
==Rodent Genomes==<br />
<br />
rat genome: 2.87 Gb<br />
<br />
rat chromosomes: 21 pairs<br />
<br />
mouse genome: 2.73 Gb<br />
<br />
mouse chromosomes: 20 pairs<br />
<br />
==Nucleosomes==<br />
<br />
Approximately 30 million nucleosomes per human cell (~ 1 / 200 bp)<br />
<br />
147 bp wrapped (1.67 left-handed superhelical turns) around histone octamer (2 copies each H2A, H2B, H3, and H4). <br />
<br />
The nucleosome contains over 120 direct protein-DNA interactions and several hundred water-mediated ones.<br />
<br />
Linker DNA between nucleosomes 10-80 bp<br />
<br />
ideal MNase mononucleosomal digest -> 3 x 10<sup>7</sup> fragments per cell (3 x 10<sup>13</sup> for 1 million cells); if 4 x 10<sup>5</sup> transcription start sites (TSS), then ~ 0.1% of mononucleosomal fragments will be of interest.<br />
<br />
Acetylation sites: "On average, 5 to 9 lysine residues of H2B, H3, and H4 were acetylated while a higher number (10 to 11 sites) of modifications was observed for H2A [but may be more]" PMID 37011222 ; so total of 50 - 76 acetylation sites per nucleosome.</div>Houpthttps://wiki.houptlab.org/index.php?title=DNA_Facts&diff=3299DNA Facts2025-07-07T17:43:26Z<p>Houpt: reformatting</p>
<hr />
<div><br />
Some useful facts about DNA.<br />
<br />
mass of DNA: 1 bp 660 Daltons (660 g/mol)<br />
<br />
molarity of 1 ug/ul of single base-pairs = 1.5 mM (divide by fragment length to get molarity of fragments)<br />
<br />
DNA per mammalian cell: 6 pg<br />
<br />
length of DNA (diploid human) per cell: 2 m<br />
<br />
==Human Genome==<br />
<br />
human genome: 3.1 Gb, so 6 billion bp in normal diploid<br />
<br />
human chromosomes: 23 pairs (22 automosomes and X and Y chromosomes) <br />
<br />
human mitochondria genome: 16.5 kbp, encodes 13 proteins; 1% of cellular DNA (1000 - 10,000 copies per cell)<br />
<br />
<br />
==Rodent Genomes==<br />
<br />
rat genome: 2.87 Gb<br />
<br />
rat chromosomes: 21 pairs<br />
<br />
mouse genome: 2.73 Gb<br />
<br />
mouse chromosomes: 20 pairs<br />
<br />
==Nucleosomes==<br />
<br />
147 bp wrapped (1.67 left-handed superhelical turns) around histone octamer ( 2 copies each H2A, H2B, H3, and H4). The nucleosome contains over 120 direct protein-DNA interactions and several hundred water-mediated ones. Linker DNA between nucleosomes 10-80 bp</div>Houpthttps://wiki.houptlab.org/index.php?title=DNA_Facts&diff=3298DNA Facts2025-07-07T17:41:26Z<p>Houpt: added a molarity calculation</p>
<hr />
<div><br />
Some useful facts about DNA.<br />
<br />
mass of DNA: 1 bp 660 Daltons (660 g/mol)<br />
<br />
molarity of 1 ug/ul of single base-pairs = 1.5 mM (divide by fragment length to get molarity of fragments)<br />
<br />
DNA per mammalian cell: 6 pg<br />
<br />
length of DNA (diploid human) per cell: 2 m<br />
<br />
human genome: 3.1 Gb, so 6 billion bp in normal diploid<br />
<br />
rat genome: 2.87 Gb<br />
<br />
mouse genome: 2.73 Gb<br />
<br />
human mitochondria genome: 16.5 kbp, encodes 13 proteins; 1% of cellular DNA (1000 - 10,000 copies per cell)<br />
<br />
human chromosomes: 23 pairs: (22 automosomes and X and Y chromosomes) <br />
<br />
rat chromosomes: 21 pairs<br />
<br />
mouse chromosomes: 20 pairs<br />
<br />
nucleosome: 147 bp wrapped (1.67 left-handed superhelical turns) around histone octamer ( 2 copies each H2A, H2B, H3, and H4). The nucleosome contains over 120 direct protein-DNA interactions and several hundred water-mediated ones. Linker DNA between nucleosomes 10-80 bp</div>Houpthttps://wiki.houptlab.org/index.php?title=DNA_Facts&diff=3297DNA Facts2025-07-07T17:38:00Z<p>Houpt: created page</p>
<hr />
<div><br />
Some useful facts about DNA.<br />
<br />
mass of DNA: 1 bp 660 Daltons (660 g/mol)<br />
<br />
DNA per mammalian cell: 6 pg<br />
<br />
length of DNA (diploid human) per cell: 2 m<br />
<br />
human genome: 3.1 Gb, so 6 billion bp in normal diploid<br />
<br />
rat genome: 2.87 Gb<br />
<br />
mouse genome: 2.73 Gb<br />
<br />
human mitochondria genome: 16.5 kbp, encodes 13 proteins; 1% of cellular DNA (1000 - 10,000 copies per cell)<br />
<br />
human chromosomes: 23 pairs: (22 automosomes and X and Y chromosomes) <br />
<br />
rat chromosomes: 21 pairs<br />
<br />
mouse chromosomes: 20 pairs<br />
<br />
nucleosome: 147 bp wrapped (1.67 left-handed superhelical turns) around histone octamer ( 2 copies each H2A, H2B, H3, and H4). The nucleosome contains over 120 direct protein-DNA interactions and several hundred water-mediated ones. Linker DNA between nucleosomes 10-80 bp</div>Houpthttps://wiki.houptlab.org/index.php?title=Fluorescent_Microscopy&diff=3296Fluorescent Microscopy2025-06-23T14:37:31Z<p>Houpt: added olympus equivalents</p>
<hr />
<div>Here are the immunofluorescence filters that we probably need. Bolded are the fluorophores we frequently use.<br />
<tab sep=tab border=1><br />
Probe Excitation (nm) Emission (nm) Color <br />
Neurotrace 435 455 blue<br />
DAPI 358 461 blue (often use mixed in [https://vectorlabs.com/products/vectashieldr-plus-antifade-mounting-medium-dapi VectaShield])<br />
'''DyLight 405''' 400 421 blue<br />
Brilliant Violet 421 407 421 blue (expensive)<br />
'''GFP''' 425 500 green<br />
'''Alexa Fluor 488''' 493 519 green<br />
Fluorescein (FITC) 492 520 green<br />
YFP 500 535 yellow<br />
Cyanine Cy3 550 570 orange<br />
Alexa Fluor 568 577 602 red<br />
Texas Red 589 615 red<br />
'''Alexa Fluor 594''' 591 614 red (sub for Texas Red)<br />
Cyanine Cy5 650 670 red<br />
Alexa Fluor 647 red, not for mercury lamps<br />
</tab><br />
<br />
==Filters in Olympus AX70 Provis Microscope==<br />
<br />
Olympus equivalents given, because that's how the cubes are labeled on the AX70 panel.<br />
<br />
2025-4-14 Purchased from [https://www.avr-optics.com AVR Optics]<br />
<br />
<br />
* [https://www.avr-optics.com/catalog/optical_filters/filter_sets/bfp_a_basic BFP-A-BASIC-000] BrightLine Basic™ single-band filter set, optimized for Blue Fluorescent Protein and other like fluorophores (roughly equivalent to Olympus U-MWBV)<br />
<br />
* [https://www.avr-optics.com/catalog/optical_filters/filter_sets/gfp_a_basic GFP-A-BASIC-000] BrightLine Basic™ single-band filter set, optimized for Green Fluorescent Protein and other like fluorophores (roughly equivalent to Olympus U-MWB)<br />
<br />
* [https://www.avr-optics.com/catalog/optical_filters/filter_sets/txred_4040c TXRED-4040C-000] BrightLine® single-band filter set, optimized for Texas Red and other like fluorophores (roughly equivalent to Olympus U-MNG)<br />
<br />
<br />
==Supplies==<br />
<br />
[https://vectorlabs.com/products/vectashield-plus-antifade VECTASHIELD® PLUS Antifade Mounting Medium (H-1900)]<br />
<br />
[https://vectorlabs.com/products/vectashieldr-plus-antifade-mounting-medium-dapi VECTASHIELD® PLUS Antifade Mounting Medium with DAPI (H-2000)]</div>Houpthttps://wiki.houptlab.org/index.php?title=DMZ_Orchiectomy_and_Butyrate_3&diff=3295DMZ Orchiectomy and Butyrate 32025-06-23T14:09:45Z<p>Houpt: moved time back 1 week</p>
<hr />
<div>Third cohort to test of role of testicular hormones in butyrate-enhanced CTA, follow-up to [[DMX Orchiectomy and Butyrate-Enhanced CTA]] and [[DMY Orchiectomy and Butyrate 2]].<br />
<br />
Mukherjee showed that HDAC inhibitors butyrate and calproate enchanced CTA in male but not female rats. Ovariectomy did not make female rats sensitive, so testing if orchiectomy reduces sensitivity of male rats.<br />
<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Orchiectomy and Butyrate 3<br />
| project = HDAC Sex Difference<br />
| code = DMZ<br />
| protocol = 202200000006<br />
| funding = Chair's Fund<br />
| investigators = Mukeherjee, Bass, Houpt<br />
| species = rat<br />
| date = 2025-6-15<br />
| type = CTA<br />
| drugs = butyrate, lithium<br />
| artifacts = <br />
| data = https://houptlab.org/bartab/expt/?id=DMZ<br />
}}<br />
<br />
Dose of NaB based on [https://houptlab.org/papers/pdfs/Houpt065.pdf Kwon and Houpt, Brain Research, 1333 (2010) 36–47], PMID 20346924 [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2871962/ PMCID: PMC2871962]<br />
<br />
* 0.15 M LiCl = 3.18g / 500ml water<br />
* 0.3 M Na Butyrate = 3.3g/100ml water (Sigma 303410-100g, pcode 102622007, source BCCJ6385)<br />
* 0.3 M NaCl = 1.8g/100ml water (osmotic control)<br />
<br />
<br><br />
<br />
==Data== <br />
<br />
'''[https://houptlab.org/bartab/expt/?id=DMZ Raw Data Link]'''<br />
<br />
==People== <br />
<br />
Mukherjee, Bass, Houpt<br />
<br />
==Agenda==<br />
<br />
'''Rats arrive''' <br />
Male Sprague Dawley rats n = 16, d.o.b. 2025-3-27, arrived: 2025-5-27<br />
<br />
<br><br />
'''Surgery'''<br />
<br />
<br />
<br />
* DMZ 1-8: Orchiectomy Surgery on 2025-6-15<br />
* DMZ 9-16: Sham Surgery on 2025-6-19<br />
<br />
<br />
Orchiectomy as per Sophocles and Idris 2019 PMID 30729469<br />
<br />
Sham rats received incision and staples.<br />
<br />
Carpofen 0.5 mg/kg pre-op, isoflurane and bupivicaine intra-op, carpofen 0.5 mg/kg post-op (next day). Staples removed 7 days post-op. <br />
<br />
<br />
'''Day 0: Water removed 2025-6-29'''<br />
<br />
<br><br />
<br />
'''Days 1-6: Water Restriction 2025-7-5 thru 2025-7-12'''<br />
<br />
''note: this would be sooner than last 2 experiments -- consider pushing back a week''<br />
<br />
Weigh rats daily. Ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily. <br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 7: Conditioning Day 2025-7-13'''<br />
<br />
* 0-30 min: saccharin 0.125%<br />
<br />
* at 30 min: 3 ml/kg of 0.15M LiCl ip (all rats)<br />
<br />
* at 40 min: 12 ml/kg of 0.3M NaButyrate or 0.3M NaCl<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
Groups:<br />
<br />
<br />
* ORC + LiCl + NaCl: DMZ01-04<br />
* ORC + LiCl + Butyrate: DMZ05-08<br />
* Sham + LiCl + NaCl: DMZ09-12<br />
* Sham + LiCl + Butyrate: DMY13-16<br />
<br />
<br><br />
<br />
'''Days 8 - 15: Preference Tests: 2025-7-14 to 2025-7-24'''<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests (for 10 days). Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]<br />
[[Category:Epigenetics]]</div>Houpthttps://wiki.houptlab.org/index.php?title=DMZ_Orchiectomy_and_Butyrate_3&diff=3294DMZ Orchiectomy and Butyrate 32025-06-20T20:46:50Z<p>Houpt: added tentative dates</p>
<hr />
<div>Third cohort to test of role of testicular hormones in butyrate-enhanced CTA, follow-up to [[DMX Orchiectomy and Butyrate-Enhanced CTA]] and [[DMY Orchiectomy and Butyrate 2]].<br />
<br />
Mukherjee showed that HDAC inhibitors butyrate and calproate enchanced CTA in male but not female rats. Ovariectomy did not make female rats sensitive, so testing if orchiectomy reduces sensitivity of male rats.<br />
<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Orchiectomy and Butyrate 3<br />
| project = HDAC Sex Difference<br />
| code = DMZ<br />
| protocol = 202200000006<br />
| funding = Chair's Fund<br />
| investigators = Mukeherjee, Bass, Houpt<br />
| species = rat<br />
| date = 2025-6-15<br />
| type = CTA<br />
| drugs = butyrate, lithium<br />
| artifacts = <br />
| data = https://houptlab.org/bartab/expt/?id=DMZ<br />
}}<br />
<br />
Dose of NaB based on [https://houptlab.org/papers/pdfs/Houpt065.pdf Kwon and Houpt, Brain Research, 1333 (2010) 36–47], PMID 20346924 [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2871962/ PMCID: PMC2871962]<br />
<br />
* 0.15 M LiCl = 3.18g / 500ml water<br />
* 0.3 M Na Butyrate = 3.3g/100ml water (Sigma 303410-100g, pcode 102622007, source BCCJ6385)<br />
* 0.3 M NaCl = 1.8g/100ml water (osmotic control)<br />
<br />
<br><br />
<br />
==Data== <br />
<br />
'''[https://houptlab.org/bartab/expt/?id=DMZ Raw Data Link]'''<br />
<br />
==People== <br />
<br />
Mukherjee, Bass, Houpt<br />
<br />
==Agenda==<br />
<br />
'''Rats arrive''' <br />
Male Sprague Dawley rats n = 16, d.o.b. 2025-3-27, arrived: 2025-5-27<br />
<br />
<br><br />
'''Surgery'''<br />
<br />
<br />
<br />
* DMZ 1-8: Orchiectomy Surgery on 2025-6-15<br />
* DMZ 9-16: Sham Surgery on 2025-6-19<br />
<br />
<br />
Orchiectomy as per Sophocles and Idris 2019 PMID 30729469<br />
<br />
Sham rats received incision and staples.<br />
<br />
Carpofen 0.5 mg/kg pre-op, isoflurane and bupivicaine intra-op, carpofen 0.5 mg/kg post-op (next day). Staples removed 7 days post-op. <br />
<br />
<br />
'''Day 0: Water removed 2025-6-29'''<br />
<br />
<br><br />
<br />
'''Days 1-6: Water Restriction 2025-6-30 thru 2025-7-5'''<br />
<br />
''note: this would be sooner than last 2 experiments -- consider pushing back a week''<br />
<br />
Weigh rats daily. Ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily. <br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 7: Conditioning Day 2025-7-6'''<br />
<br />
* 0-30 min: saccharin 0.125%<br />
<br />
* at 30 min: 3 ml/kg of 0.15M LiCl ip (all rats)<br />
<br />
* at 40 min: 12 ml/kg of 0.3M NaButyrate or 0.3M NaCl<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
Groups:<br />
<br />
<br />
* ORC + LiCl + NaCl: DMZ01-04<br />
* ORC + LiCl + Butyrate: DMZ05-08<br />
* Sham + LiCl + NaCl: DMZ09-12<br />
* Sham + LiCl + Butyrate: DMY13-16<br />
<br />
<br><br />
<br />
'''Days 8 - 15: Preference Tests: 2025-7-7 to 2025-7-14'''<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests (for 7 days). Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 7 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]<br />
[[Category:Epigenetics]]</div>Houpthttps://wiki.houptlab.org/index.php?title=DMZ_Orchiectomy_and_Butyrate_3&diff=3293DMZ Orchiectomy and Butyrate 32025-06-17T14:15:24Z<p>Houpt: added dob</p>
<hr />
<div>Third cohort to test of role of testicular hormones in butyrate-enhanced CTA, follow-up to [[DMX Orchiectomy and Butyrate-Enhanced CTA]] and [[DMY Orchiectomy and Butyrate 2]].<br />
<br />
Mukherjee showed that HDAC inhibitors butyrate and calproate enchanced CTA in male but not female rats. Ovariectomy did not make female rats sensitive, so testing if orchiectomy reduces sensitivity of male rats.<br />
<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Orchiectomy and Butyrate 3<br />
| project = HDAC Sex Difference<br />
| code = DMZ<br />
| protocol = 202200000006<br />
| funding = Chair's Fund<br />
| investigators = Mukeherjee, Bass, Houpt<br />
| species = rat<br />
| date = 2025-6-15<br />
| type = CTA<br />
| drugs = butyrate, lithium<br />
| artifacts = <br />
| data = https://houptlab.org/bartab/expt/?id=DMZ<br />
}}<br />
<br />
Dose of NaB based on [https://houptlab.org/papers/pdfs/Houpt065.pdf Kwon and Houpt, Brain Research, 1333 (2010) 36–47], PMID 20346924 [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2871962/ PMCID: PMC2871962]<br />
<br />
* 0.15 M LiCl = 3.18g / 500ml water<br />
* 0.3 M Na Butyrate = 3.3g/100ml water (Sigma 303410-100g, pcode 102622007, source BCCJ6385)<br />
* 0.3 M NaCl = 1.8g/100ml water (osmotic control)<br />
<br />
<br><br />
<br />
==Data== <br />
<br />
'''[https://houptlab.org/bartab/expt/?id=DMZ Raw Data Link]'''<br />
<br />
==People== <br />
<br />
Mukherjee, Bass, Houpt<br />
<br />
==Agenda==<br />
<br />
'''Rats arrive''' <br />
Male Sprague Dawley rats n = 16, d.o.b. 2025-3-27, arrived: 2025-5-27<br />
<br />
<br><br />
'''Surgery'''<br />
<br />
<br />
<br />
* DMZ 1-8: Orchiectomy Surgery on 2025-6-15<br />
* DMZ 9-16: Sham Surgery on 2025-6<br />
<br />
<br />
Orchiectomy as per Sophocles and Idris 2019 PMID 30729469<br />
<br />
Sham rats received incision and staples.<br />
<br />
Carpofen 0.5 mg/kg pre-op, isoflurane and bupivicaine intra-op, carpofen 0.5 mg/kg post-op (next day). Staples removed 7 days post-op. <br />
<br />
<br />
'''Day 0: Water removed 2025-'''<br />
<br />
<br><br />
<br />
'''Days 1-6: Water Restriction XXX thru XXX''<br />
<br />
Weigh rats daily. Ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily. <br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 7: Conditioning Day XXXXX'''<br />
<br />
* 0-30 min: saccharin 0.125%<br />
<br />
* at 30 min: 3 ml/kg of 0.15M LiCl ip (all rats)<br />
<br />
* at 40 min: 12 ml/kg of 0.3M NaButyrate or 0.3M NaCl<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
Groups:<br />
<br />
<br />
* ORC + LiCl + NaCl: DMZ01-04<br />
* ORC + LiCl + Butyrate: DMZ05-08<br />
* Sham + LiCl + NaCl: DMZ09-12<br />
* Sham + LiCl + Butyrate: DMY13-16<br />
<br />
<br><br />
<br />
'''Days 8 - ???: Preference Tests: XXXX to XXXX'''<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests (for ???? days). Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for ??? days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]<br />
[[Category:Epigenetics]]</div>Houpthttps://wiki.houptlab.org/index.php?title=DMY_Orchiectomy_and_Butyrate_2&diff=3292DMY Orchiectomy and Butyrate 22025-06-16T14:51:46Z<p>Houpt: updated length of preference tests</p>
<hr />
<div>Second cohort to test of role of testicular hormones in butyrate-enhanced CTA, follow-up to [[DMX Orchiectomy and Butyrate-Enhanced CTA]].<br />
<br />
Mukherjee showed that HDAC inhibitors butyrate and calproate enchanced CTA in male but not female rats. Ovariectomy did not make female rats sensitive, so testing if orchiectomy reduces sensitivity of male rats.<br />
<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Orchiectomy and Butyrate 2<br />
| project = HDAC Sex Difference<br />
| code = DMY<br />
| protocol = 202200000006<br />
| funding = Chair's Fund<br />
| investigators = Mukeherjee, Bass, Houpt<br />
| species = rat<br />
| date = 2024-8-6<br />
| type = CTA<br />
| drugs = butyrate, lithium<br />
| artifacts = <br />
| data = https://houptlab.org/bartab/expt/?id=DMY<br />
}}<br />
<br />
Dose of NaB based on [https://houptlab.org/papers/pdfs/Houpt065.pdf Kwon and Houpt, Brain Research, 1333 (2010) 36–47], PMID 20346924 [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2871962/ PMCID: PMC2871962]<br />
<br />
* 0.15 M LiCl = 3.18g / 500ml water<br />
* 0.3 M Na Butyrate = 3.3g/100ml water (Sigma 303410-100g, pcode 102622007, source BCCJ6385)<br />
* 0.3 M NaCl = 1.8g/100ml water (osmotic control)<br />
<br />
<br><br />
<br />
==Data== <br />
<br />
'''[https://houptlab.org/bartab/expt/?id=DMY Raw Data Link]'''<br />
<br />
==People== <br />
<br />
Mukherjee, Bass, Houpt<br />
<br />
==Agenda==<br />
<br />
'''Rats arrive''' <br />
Male Sprague Dawley rats n = 16, d.o.b. 2024-6-10, arrived: 2024-8-6<br />
<br />
<br><br />
'''Surgery'''<br />
<br />
<br />
<br />
* DMY 1-8: Orchiectomy Surgery on 2024-9-1<br />
* DMY 9-15: Sham Surgery on 2024-9-7<br />
* DMY 16: Orchiectomy Surgery on 2024-9-7<br />
(NB: DMY16 relabeled DMY8 on 2024-10-6)<br />
<br />
Orchiectomy as per Sophocles and Idris 2019 PMID 30729469<br />
<br />
Sham rats received incision and staples.<br />
<br />
Carpofen 0.5 mg/kg pre-op, isoflurane and bupivicaine intra-op, carpofen 0.5 mg/kg post-op (next day). Staples removed 7 days post-op. <br />
<br />
<br />
'''Day 0: Water removed 2024-10-6'''<br />
<br />
<br><br />
<br />
'''Days 1-6: Water Restriction 2024-10-7 thru 2024-10-12''<br />
<br />
Weigh rats daily. Ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily. <br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 7: Conditioning Day 2024-10-13'''<br />
<br />
* 0-30 min: saccharin 0.125%<br />
<br />
* at 30 min: 3 ml/kg of 0.15M LiCl ip (all rats)<br />
<br />
* at 40 min: 12 ml/kg of 0.3M NaButyrate or 0.3M NaCl<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
Groups:<br />
<br />
<br />
* ORC + LiCl + NaCl: DMY01-04<br />
* ORC + LiCl + Butyrate: DMY05-08<br />
* Sham + LiCl + NaCl: DMY09-11<br />
* Sham + LiCl + Butyrate: DMY12-15<br />
<br />
<br><br />
<br />
'''Days 8 - 14: Preference Tests: 2024-10-14 to 2024-10-21'''<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests (for 7 days). Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 7 days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]<br />
[[Category:Epigenetics]]</div>Houpthttps://wiki.houptlab.org/index.php?title=DMZ_Orchiectomy_and_Butyrate_3&diff=3291DMZ Orchiectomy and Butyrate 32025-06-16T14:50:14Z<p>Houpt: created page</p>
<hr />
<div>Third cohort to test of role of testicular hormones in butyrate-enhanced CTA, follow-up to [[DMX Orchiectomy and Butyrate-Enhanced CTA]] and [[DMY Orchiectomy and Butyrate 2]].<br />
<br />
Mukherjee showed that HDAC inhibitors butyrate and calproate enchanced CTA in male but not female rats. Ovariectomy did not make female rats sensitive, so testing if orchiectomy reduces sensitivity of male rats.<br />
<br />
<br />
{{Infobox experiment|align=right<br />
| Name = Orchiectomy and Butyrate 3<br />
| project = HDAC Sex Difference<br />
| code = DMZ<br />
| protocol = 202200000006<br />
| funding = Chair's Fund<br />
| investigators = Mukeherjee, Bass, Houpt<br />
| species = rat<br />
| date = 2025-6-15<br />
| type = CTA<br />
| drugs = butyrate, lithium<br />
| artifacts = <br />
| data = https://houptlab.org/bartab/expt/?id=DMZ<br />
}}<br />
<br />
Dose of NaB based on [https://houptlab.org/papers/pdfs/Houpt065.pdf Kwon and Houpt, Brain Research, 1333 (2010) 36–47], PMID 20346924 [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2871962/ PMCID: PMC2871962]<br />
<br />
* 0.15 M LiCl = 3.18g / 500ml water<br />
* 0.3 M Na Butyrate = 3.3g/100ml water (Sigma 303410-100g, pcode 102622007, source BCCJ6385)<br />
* 0.3 M NaCl = 1.8g/100ml water (osmotic control)<br />
<br />
<br><br />
<br />
==Data== <br />
<br />
'''[https://houptlab.org/bartab/expt/?id=DMZ Raw Data Link]'''<br />
<br />
==People== <br />
<br />
Mukherjee, Bass, Houpt<br />
<br />
==Agenda==<br />
<br />
'''Rats arrive''' <br />
Male Sprague Dawley rats n = 16, d.o.b. 2024-6-10, arrived: 2024-8-6<br />
<br />
<br><br />
'''Surgery'''<br />
<br />
<br />
<br />
* DMZ 1-8: Orchiectomy Surgery on 2025-6-15<br />
* DMZ 9-16: Sham Surgery on 2025-6<br />
<br />
<br />
Orchiectomy as per Sophocles and Idris 2019 PMID 30729469<br />
<br />
Sham rats received incision and staples.<br />
<br />
Carpofen 0.5 mg/kg pre-op, isoflurane and bupivicaine intra-op, carpofen 0.5 mg/kg post-op (next day). Staples removed 7 days post-op. <br />
<br />
<br />
'''Day 0: Water removed 2025-'''<br />
<br />
<br><br />
<br />
'''Days 1-6: Water Restriction XXX thru XXX''<br />
<br />
Weigh rats daily. Ad lib food. <br />
<br />
30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1400 daily. <br />
<br />
1 water bottle, 1 empty bottle to accustom to 2-bottle choice<br />
<br />
<br><br />
<br />
'''Day 7: Conditioning Day XXXXX'''<br />
<br />
* 0-30 min: saccharin 0.125%<br />
<br />
* at 30 min: 3 ml/kg of 0.15M LiCl ip (all rats)<br />
<br />
* at 40 min: 12 ml/kg of 0.3M NaButyrate or 0.3M NaCl<br />
<br />
ad lib water returned after final injection overnight.<br />
<br />
Groups:<br />
<br />
<br />
* ORC + LiCl + NaCl: DMZ01-04<br />
* ORC + LiCl + Butyrate: DMZ05-08<br />
* Sham + LiCl + NaCl: DMZ09-12<br />
* Sham + LiCl + Butyrate: DMY13-16<br />
<br />
<br><br />
<br />
'''Days 8 - ???: Preference Tests: XXXX to XXXX'''<br />
<br />
Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests (for ???? days). Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for ??? days.<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]<br />
[[Category:Epigenetics]]</div>Houpthttps://wiki.houptlab.org/index.php?title=Adrenal_Gland&diff=3284Adrenal Gland2025-04-25T13:52:57Z<p>Houpt: created page</p>
<hr />
<div><br />
Some papers on [https://houptlab.org/courses/adrenal adrenal gland and regulation of the medulla].<br />
<br />
=Rat Adrenal Gland Facts=<br />
<br />
''From Tomlinson 1987 PMID 3601067''<br />
<br />
Adrenal Gland: 13.2 mm<sup>3</sup><br />
<br />
Adrenal Medulla: 1.3 mm<sup>3</sup><br />
<br />
Medulla by volume: 63% chromaffin (catecholamine) cells, 20% vasculature, 5% neural, 12% interstitial<br />
<br />
Number of cells (unilateral):<br />
<br />
* 5 x10<sup>5</sup> epinephrine cells (TH & PNMT)<br />
<br />
* 2 x 10<sup>5</sup> norepinephrine cells (TH)</div>Houpthttps://wiki.houptlab.org/index.php?title=Fluorescent_Microscopy&diff=3283Fluorescent Microscopy2025-04-14T19:55:19Z<p>Houpt: added new filters</p>
<hr />
<div>Here are the immunofluorescence filters that we probably need. Bolded are the fluorophores we frequently use.<br />
<tab sep=tab border=1><br />
Probe Excitation (nm) Emission (nm) Color <br />
Neurotrace 435 455 blue<br />
DAPI 358 461 blue (often use mixed in [https://vectorlabs.com/products/vectashieldr-plus-antifade-mounting-medium-dapi VectaShield])<br />
'''DyLight 405''' 400 421 blue<br />
Brilliant Violet 421 407 421 blue (expensive)<br />
'''GFP''' 425 500 green<br />
'''Alexa Fluor 488''' 493 519 green<br />
Fluorescein (FITC) 492 520 green<br />
YFP 500 535 yellow<br />
Cyanine Cy3 550 570 orange<br />
Alexa Fluor 568 577 602 red<br />
Texas Red 589 615 red<br />
'''Alexa Fluor 594''' 591 614 red (sub for Texas Red)<br />
Cyanine Cy5 650 670 red<br />
Alexa Fluor 647 red, not for mercury lamps<br />
</tab><br />
<br />
==Filters in Olympus AX70 Provis Microscope==<br />
<br />
2025-4-14 Purchased from [https://www.avr-optics.com AVR Optics]<br />
<br />
<br />
* [https://www.avr-optics.com/catalog/optical_filters/filter_sets/bfp_a_basic BFP-A-BASIC-000] BrightLine Basic™ single-band filter set, optimized for Blue Fluorescent Protein and other like fluorophores<br />
<br />
* [https://www.avr-optics.com/catalog/optical_filters/filter_sets/gfp_a_basic GFP-A-BASIC-000] BrightLine Basic™ single-band filter set, optimized for Green Fluorescent Protein and other like fluorophores<br />
<br />
* [https://www.avr-optics.com/catalog/optical_filters/filter_sets/txred_4040c TXRED-4040C-000] BrightLine® single-band filter set, optimized for Texas Red and other like fluorophores<br />
<br />
<br />
==Supplies==<br />
<br />
[https://vectorlabs.com/products/vectashield-plus-antifade VECTASHIELD® PLUS Antifade Mounting Medium (H-1900)]<br />
<br />
[https://vectorlabs.com/products/vectashieldr-plus-antifade-mounting-medium-dapi VECTASHIELD® PLUS Antifade Mounting Medium with DAPI (H-2000)]</div>Houpthttps://wiki.houptlab.org/index.php?title=C-Fos_Antibodies&diff=3281C-Fos Antibodies2025-02-19T20:57:54Z<p>Houpt: /* Santa Cruz C-10 */ add fluorescent dilutions</p>
<hr />
<div>{{lowercase title}}<br />
<br />
==Santa Cruz C-10==<br />
<br />
[https://www.scbt.com/p/c-fos-antibody-c-10 mouse anti-c-Fos antibody (C-10; Santa Cruz)] at a dilution of 1:1000 for DAB staining.<br />
<br />
2025-2-19 For immunofluoresence: C-10 at 1:200 or 1:100 (better) and Alexa 594 (red) secondary at dilution of 1:200. Still a little dim, but can easily see c-Fos-positive nuclei in e.g. CeA after LiCl.<br />
<br />
==Santa Cruz sc-52==<br />
<br />
NO LONGER IN PRODUCTION<br />
<br />
anti-c-Fos rabbit polyclonal IgG, catalog [https://www.scbt.com/p/c-fos-antibody-4 no. sc-52], 200 µg/ml, $259 <br />
<br />
also available as goat IgG, 200 µg/ml, sc-52-G, see below<br />
<br />
datasheet: http://datasheets.scbt.com/sc-52.pdf<br />
<br />
Epitope: mapping at the N-terminus of c-Fos of human origin<br />
<br />
From Santa Cruz Biotech:<br />
<tt><br />
:Dear Dr. Tom Houpt,<br />
<br />
:Greetings from Santa Cruz Biotechnology.<br />
<br />
:The exact immunogen used is proprietary, however I would be more than happy to provide you with an 50 amino acid range. The 50 amino acid range corresponds to amino acids 1-50 of the published protein accession P01100.<br />
<br />
:If you have any further questions, please feel free to email me.<br />
<br />
:Sincerely,<br />
<br />
:Loren Hatt, Technical Service Representative, Santa Cruz Biotechnology, Inc.<br />
<br />
:(800) 457-3801, h-a-t-t-l@scbt.com<br />
</tt><br />
<br />
==Calbiochem Ab-5==<br />
<br />
Anti-c-Fos (Ab-5) (4-17) Rabbit pAb, catalog [http://www.emdchemicals.com/life-science-research/products/EMD_BIO-PC38/p_uuid No. PC38] 100 µl, $213. [http://www.emdchemicals.com/is-bin/INTERSHOP.enfinity/WFS/Merck-US-Site/en_CA/-/USD/ViewProductDetail-UserProtocolDataSheet?ProductUUID=E_ab.s1Oh2EAAAEiRT1RV1.J&PortalCatalogUUID=4_6b.s1OricAAAEg3cd6cjOt&SelectedDocumentType=UserProtocolDataSheet Data sheet].<br />
<br />
Epitope: A synthetic peptide (SGFNADYEASSSRC) (Cat. No. PP10) corresponding to amino acids 4-17 of human c-Fos<br />
<br />
==Santa Cruz 4-G==<br />
<br />
NO LONGER IN PRODUCTION<br />
<br />
Anti-c-Fos (4G) goat polyclonal, catalog [https://www.scbt.com/p/c-fos-antibody-4 SC-52-G] , $279<br />
<br />
data sheet: http://datasheets.scbt.com/sc-52.pdf<br />
<br />
Dilution: use at 1:1000 for DAB chromogenic IHC, based on dilution curve using LiCl-induced c-Fos in rat forebrain from 2012-1-10.<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:c-Fos]]</div>Houpthttps://wiki.houptlab.org/index.php?title=Fluorescent_Microscopy&diff=3280Fluorescent Microscopy2025-02-05T19:12:25Z<p>Houpt: added 488, and vectashield links</p>
<hr />
<div>Here are the immunofluorescence filters that we probably need. Bolded are the fluorophores we frequently use.<br />
<tab sep=tab border=1><br />
Probe Excitation (nm) Emission (nm) Color <br />
Neurotrace 435 455 blue<br />
DAPI 358 461 blue (often use mixed in [https://vectorlabs.com/products/vectashieldr-plus-antifade-mounting-medium-dapi VectaShield])<br />
'''DyLight 405''' 400 421 blue<br />
Brilliant Violet 421 407 421 blue (expensive)<br />
'''GFP''' 425 500 green<br />
'''Alexa Fluor 488''' 493 519 green<br />
Fluorescein (FITC) 492 520 green<br />
YFP 500 535 yellow<br />
Cyanine Cy3 550 570 orange<br />
Alexa Fluor 568 577 602 red<br />
Texas Red 589 615 red<br />
'''Alexa Fluor 594''' 591 614 red (sub for Texas Red)<br />
Cyanine Cy5 650 670 red<br />
Alexa Fluor 647 red, not for mercury lamps<br />
</tab><br />
<br />
[https://vectorlabs.com/products/vectashield-plus-antifade VECTASHIELD® PLUS Antifade Mounting Medium (H-1900)]<br />
<br />
[https://vectorlabs.com/products/vectashieldr-plus-antifade-mounting-medium-dapi VECTASHIELD® PLUS Antifade Mounting Medium with DAPI (H-2000)]</div>Houpthttps://wiki.houptlab.org/index.php?title=Fluorescent_Microscopy&diff=3279Fluorescent Microscopy2025-02-05T14:54:48Z<p>Houpt: added some more</p>
<hr />
<div>Here are the immunofluorescence filters that we probably need.<br />
<tab sep=tab border=1><br />
Probe Excitation (nm) Emission (nm) Color <br />
Neurotrace 435 455 blue<br />
DAPI 358 461 blue<br />
'''DyLight 405''' 400 421 blue<br />
Brilliant Violet 421 407 421 blue (expensive)<br />
'''GFP''' 425 500 green<br />
'''Fluorescein (FITC)''' 492 520 green<br />
YFP 500 535 yellow<br />
Cyanine Cy3 550 570 orange<br />
Alexa Fluor 568 577 602 red<br />
Texas Red 589 615 red<br />
'''Alexa Fluor 594''' 591 614 red (sub for Texas Red)<br />
Cyanine Cy5 650 670 red<br />
Alexa Fluor 647 red, not for mercury lamps<br />
</tab></div>Houpthttps://wiki.houptlab.org/index.php?title=GFA_phospho-PDH_and_Taste_Aversion&diff=3278GFA phospho-PDH and Taste Aversion2025-02-04T20:08:24Z<p>Houpt: added c-Fos note</p>
<hr />
<div><br />
Examine levels of [[Phospho-Pyruvate Dehydrogenase (pPDH)]] by immunofluorescence in after intraoral conditioned taste aversion.<br />
<br />
Based on Yang et al. 2024 PMID 38266644 and report by Roitman lab at SSIB 2024.<br />
<br />
Employed SD-GFP-TH rats (Iacovitti et al 2014 PMID 25462571) from Taconic that express GFP under TH promoter with especially good expression in VTA. <br />
<br />
For immunohistochemistry:<br />
<br />
* for pPDH: [https://www.cellsignal.com/products/primary-antibodies/phospho-pyruvate-dehydrogenase-a1-ser293-e4v9l-rabbit-mab/37115 Phospho-Pyruvate Dehydrogenase α1 (Ser293) (E4V9L) Rabbit mAb #37115] at 1:500 for immunofluoresence; BLUE secondary [https://www.jacksonimmuno.com/catalog/products/711-475-152 Jackson DyLight™ 405 AffiniPure™ Donkey Anti-Rabbit IgG (H+L) #711-475-152] [WHAT IS CONCENTRATION?] <br />
<br />
* for c-Fos: [https://www.scbt.com/p/c-fos-antibody-c-10 mouse anti-c-Fos antibody (C-10; Santa Cruz)] at a dilution of 1:1000. GET Antibody specification and secondary from Jason's work<br />
<br />
<br />
{{Infobox experiment|align=right<br />
| Name = phospho-PDH and CTA<br />
| project = Taste Aversion<br />
| code = GFA<br />
| protocol = 202200000006<br />
| funding = Chair's Fund<br />
| investigators = Houpt, Cote, Trombley, Blakemore<br />
| species = rat<br />
| date = 2024-12-3<br />
| type = CTA<br />
| drugs = lithium<br />
| data = https://houptlab.org/bartab/expt/?id=GFA<br />
}}<br />
<br />
<br />
==Data==<br />
<br />
'''[https://houptlab.org/bartab/expt/?id=GFA Raw Data Link]'''<br />
<br />
<br />
==People==<br />
<br />
Jason Cote (breeding), Laura Blakemore and Paul Trombley (colony foundation and management)<br />
<br />
==CTA Conditioning==<br />
<br />
===Agenda===<br />
<br />
'''Rats Arrive''' (December 3, 2024)<br />
<br />
Rats (n=4 male, n=5 female, littermates, DOB: 7-???-2024) transferred from GFP-TH breeding protocol to individual cages.<br />
<br />
<br><br />
<br />
<br />
'''Day 0: Surgery''' (December XXX, 2024)<br />
<br />
Chronic Intraoral Catheters implanted under isoflurane anesthesia. All rats receive carprofen (XXX mg/kg) pre-operatively and 1 day post-op.<br />
<br />
<br><br />
<br />
<br />
'''Days 1 - 3: Recovery''' (December XXXX - XXXX, 2024)<br />
<br />
Weigh rats daily. Ad lib food and water. Carprofen (XXX mg/kg) on first day post-op.<br />
<br />
<br><br />
<br />
<br />
'''Days 5-7: Habituation''' (December XXXX - XXXX, 2024)<br />
<br />
Rats given intraoral infusion of 6 ml dH2O over 6 min to habituate to infusion procedure<br />
<br />
Food removed on last day of habituation.<br />
<br />
'''Noncontingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).'''<br />
<br />
<br><br />
<br />
<br />
'''Day 8: Conditioning day 1''' (December XXXX, 2024)<br />
<br />
All rats given intraoral infusion of 6 ml sucrose 5% over 6 min.<br />
<br />
'''Contingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).'''<br />
<br />
Food returned overnight.<br />
<br />
<br><br />
<br />
<br />
'''Day 9: Recovery Day''' (December XXXX, 2024)<br />
<br />
Food removed.<br />
<br />
'''Noncontingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).'''<br />
<br />
<br><br />
<br />
<br />
'''Day 10: Conditioning day 2''' (December XXXX, 2024)<br />
<br />
All rats given intraoral infusion of 6 ml sucrose 5% over 6 min.<br />
<br />
'''Contingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).'''<br />
<br />
Food returned overnight.<br />
<br />
<br><br />
<br />
<br />
'''Day 11: Recovery Day''' (December XXXX, 2024)<br />
<br />
Food removed.<br />
<br />
'''Noncontingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).'''<br />
<br />
<br><br />
<br />
<br />
'''Day 12: Conditioning day 2''' (December XXXX, 2024)<br />
<br />
All rats given intraoral infusion of 6 ml sucrose 5% over 6 min.<br />
<br />
'''Contingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).'''<br />
<br />
Food returned overnight.<br />
<br />
<br><br />
<br />
<br />
'''Day 13: Recovery Day''' (December XXXX, 2024)<br />
<br />
Food removed.<br />
<br />
<br><br />
<br />
<br />
'''Day 14: Expression Test''' (December XXXX, 2024)<br />
<br />
All rats given intraoral infusion of 6 ml sucrose 5% over 6 min.<br />
<br />
One hour after infusion, rats are overdosed with euthanasol and transcardially perfused. Brains postfixed for 1-2 hours, then stored in 0.1M PB.<br />
<br />
==Histology==<br />
<br />
<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:CTA Experiments]]</div>Houpthttps://wiki.houptlab.org/index.php?title=C-Fos_Antibodies&diff=3277C-Fos Antibodies2025-02-04T20:07:33Z<p>Houpt: added note on 4-G</p>
<hr />
<div>{{lowercase title}}<br />
<br />
==Santa Cruz C-10==<br />
<br />
[https://www.scbt.com/p/c-fos-antibody-c-10 mouse anti-c-Fos antibody (C-10; Santa Cruz)] at a dilution of 1:1000 for DAB staining.<br />
<br />
==Santa Cruz sc-52==<br />
<br />
NO LONGER IN PRODUCTION<br />
<br />
anti-c-Fos rabbit polyclonal IgG, catalog [https://www.scbt.com/p/c-fos-antibody-4 no. sc-52], 200 µg/ml, $259 <br />
<br />
also available as goat IgG, 200 µg/ml, sc-52-G, see below<br />
<br />
datasheet: http://datasheets.scbt.com/sc-52.pdf<br />
<br />
Epitope: mapping at the N-terminus of c-Fos of human origin<br />
<br />
From Santa Cruz Biotech:<br />
<tt><br />
:Dear Dr. Tom Houpt,<br />
<br />
:Greetings from Santa Cruz Biotechnology.<br />
<br />
:The exact immunogen used is proprietary, however I would be more than happy to provide you with an 50 amino acid range. The 50 amino acid range corresponds to amino acids 1-50 of the published protein accession P01100.<br />
<br />
:If you have any further questions, please feel free to email me.<br />
<br />
:Sincerely,<br />
<br />
:Loren Hatt, Technical Service Representative, Santa Cruz Biotechnology, Inc.<br />
<br />
:(800) 457-3801, h-a-t-t-l@scbt.com<br />
</tt><br />
<br />
==Calbiochem Ab-5==<br />
<br />
Anti-c-Fos (Ab-5) (4-17) Rabbit pAb, catalog [http://www.emdchemicals.com/life-science-research/products/EMD_BIO-PC38/p_uuid No. PC38] 100 µl, $213. [http://www.emdchemicals.com/is-bin/INTERSHOP.enfinity/WFS/Merck-US-Site/en_CA/-/USD/ViewProductDetail-UserProtocolDataSheet?ProductUUID=E_ab.s1Oh2EAAAEiRT1RV1.J&PortalCatalogUUID=4_6b.s1OricAAAEg3cd6cjOt&SelectedDocumentType=UserProtocolDataSheet Data sheet].<br />
<br />
Epitope: A synthetic peptide (SGFNADYEASSSRC) (Cat. No. PP10) corresponding to amino acids 4-17 of human c-Fos<br />
<br />
==Santa Cruz 4-G==<br />
<br />
NO LONGER IN PRODUCTION<br />
<br />
Anti-c-Fos (4G) goat polyclonal, catalog [https://www.scbt.com/p/c-fos-antibody-4 SC-52-G] , $279<br />
<br />
data sheet: http://datasheets.scbt.com/sc-52.pdf<br />
<br />
Dilution: use at 1:1000 for DAB chromogenic IHC, based on dilution curve using LiCl-induced c-Fos in rat forebrain from 2012-1-10.<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:c-Fos]]</div>Houpthttps://wiki.houptlab.org/index.php?title=C-Fos_Antibodies&diff=3276C-Fos Antibodies2025-02-04T20:06:57Z<p>Houpt: added C-10</p>
<hr />
<div>{{lowercase title}}<br />
<br />
==Santa Cruz C-10==<br />
<br />
[https://www.scbt.com/p/c-fos-antibody-c-10 mouse anti-c-Fos antibody (C-10; Santa Cruz)] at a dilution of 1:1000 for DAB staining.<br />
<br />
==Santa Cruz sc-52==<br />
<br />
NO LONGER IN PRODUCTION<br />
<br />
anti-c-Fos rabbit polyclonal IgG, catalog [https://www.scbt.com/p/c-fos-antibody-4 no. sc-52], 200 µg/ml, $259 <br />
<br />
also available as goat IgG, 200 µg/ml, sc-52-G, see below<br />
<br />
datasheet: http://datasheets.scbt.com/sc-52.pdf<br />
<br />
Epitope: mapping at the N-terminus of c-Fos of human origin<br />
<br />
From Santa Cruz Biotech:<br />
<tt><br />
:Dear Dr. Tom Houpt,<br />
<br />
:Greetings from Santa Cruz Biotechnology.<br />
<br />
:The exact immunogen used is proprietary, however I would be more than happy to provide you with an 50 amino acid range. The 50 amino acid range corresponds to amino acids 1-50 of the published protein accession P01100.<br />
<br />
:If you have any further questions, please feel free to email me.<br />
<br />
:Sincerely,<br />
<br />
:Loren Hatt, Technical Service Representative, Santa Cruz Biotechnology, Inc.<br />
<br />
:(800) 457-3801, h-a-t-t-l@scbt.com<br />
</tt><br />
<br />
==Calbiochem Ab-5==<br />
<br />
Anti-c-Fos (Ab-5) (4-17) Rabbit pAb, catalog [http://www.emdchemicals.com/life-science-research/products/EMD_BIO-PC38/p_uuid No. PC38] 100 µl, $213. [http://www.emdchemicals.com/is-bin/INTERSHOP.enfinity/WFS/Merck-US-Site/en_CA/-/USD/ViewProductDetail-UserProtocolDataSheet?ProductUUID=E_ab.s1Oh2EAAAEiRT1RV1.J&PortalCatalogUUID=4_6b.s1OricAAAEg3cd6cjOt&SelectedDocumentType=UserProtocolDataSheet Data sheet].<br />
<br />
Epitope: A synthetic peptide (SGFNADYEASSSRC) (Cat. No. PP10) corresponding to amino acids 4-17 of human c-Fos<br />
<br />
==Santa Cruz 4-G==<br />
<br />
Anti-c-Fos (4G) goat polyclonal, catalog [https://www.scbt.com/p/c-fos-antibody-4 SC-52-G] , $279<br />
<br />
data sheet: http://datasheets.scbt.com/sc-52.pdf<br />
<br />
Dilution: use at 1:1000 for DAB chromogenic IHC, based on dilution curve using LiCl-induced c-Fos in rat forebrain from 2012-1-10.<br />
<br />
[[Category:Taste Aversion]]<br />
[[Category:c-Fos]]</div>Houpt