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Test of role of sex in LiCl-induced and butyrate-enhanced acetylation of histones and c-Fos expression as measured by flurorescent immunohistochemistry.
Dose of NaB and LiCl and timing based on Figure 7 of [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]
First Injection:
* 0.15 M LiCl (6 ml/kg)
* 0.15 M NaCl (6 ml/kg)
Second Injection:
* 0.3 M Na Butyrate = 3.3g/100ml water (12 ml/kg)
* 0.3 M NaCl = 18g/liter water (osmotic control) (12 ml/kg)
<br>
==Data==
==People==
Durba Mukherjee
===Agenda===
'''Rats arrive''' (2022-02-23)
male (n=8) DOB: 2021-12-27
female (n=8)DOB: 2021-12-6,
<br>
'''Surgery'''
4 groups for each sex: NaCl-NaCl, LiCl-NaCl, NaCl-NaB, LiCl-NaB
Female: DMU01-DMU08; Male: DMU09-DMU16
* NaCl-NaCl: DMU 1,5,9,13
* LiCl-NaCl: Female: DMU 2,3,7; Male: DMU 10,11,15
* NaCl-NaB:
* LiCl-NaB: Female: DMU 4,6,8; Male: DMU 12,14,16
'''Injections and Perfusion '''
Rats weighed and given first injection (NaCl or LiCl). 10 minutes later, second injection (NaCl or NaB). 50 minutes later, overdosed with pentobarbital and transcardially perfused, first with 100 ml 0.15 M NaCl/NaNitrite/heparin, then 400ml of 4% PFA. Brain dissected, forebrain/brainstem blocked, post-fixed in 4% PFA for 1 h, then cryopotected in 30% sucrose for 2-3 days.
2022-2-26: DMU01-04,DMU9-12 perfused
2022-2-27: DMU05-8,DMU13-16 perfused
<br>
'''Brain Sectioning'''
36 x 40 micron sections through hindbrain (NTS)
2022-3-2: DMU01-04,DMU9-12 hindbrain sectioned
2022-3-3: DMU05-8,DMU13-16 hindbrain sectioned
'''Immunohistochemistry'''
anti-acetyl-Histone Primary: anti-acetyl-Histone H3 (Lys14) rabbit polyclonal, Millipore 07-353, Lot #3259085, r. 2019-09-26 (use at 1:2000)
Secondary: FITC conjugated Goat Anti-Rabbit, Jackson Labs 111-095-144, Lot #77909, r. 2008-05-24 (use at 1:100)
anti-c-Fos Primary: anti-c-Fos monoclonal antibody (E-8), Santa Cruz sc-166940, Lot # L1621, r. 2022-3-3 (use at 1:100)
Secondary: Texas Red conjugated Goat Anti-Mouse, Jackson Labs 115-075-146, Lot #72021, r. 2008-05-24 (use at 1:100)
[[Category:Taste Aversion]]
[[Category: CTA Experiments]]
DMU Acetyl-Histone and c-Fos Immuno (view source)
Revision as of 12:44, 3 March 2022
, 12:44, 3 March 2022created page
Test of role of sex in LiCl-induced and butyrate-enhanced acetylation of histones and c-Fos expression as measured by flurorescent immunohistochemistry.
Dose of NaB and LiCl and timing based on Figure 7 of [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]
First Injection:
* 0.15 M LiCl (6 ml/kg)
* 0.15 M NaCl (6 ml/kg)
Second Injection:
* 0.3 M Na Butyrate = 3.3g/100ml water (12 ml/kg)
* 0.3 M NaCl = 18g/liter water (osmotic control) (12 ml/kg)
<br>
==Data==
==People==
Durba Mukherjee
===Agenda===
'''Rats arrive''' (2022-02-23)
male (n=8) DOB: 2021-12-27
female (n=8)DOB: 2021-12-6,
<br>
'''Surgery'''
4 groups for each sex: NaCl-NaCl, LiCl-NaCl, NaCl-NaB, LiCl-NaB
Female: DMU01-DMU08; Male: DMU09-DMU16
* NaCl-NaCl: DMU 1,5,9,13
* LiCl-NaCl: Female: DMU 2,3,7; Male: DMU 10,11,15
* NaCl-NaB:
* LiCl-NaB: Female: DMU 4,6,8; Male: DMU 12,14,16
'''Injections and Perfusion '''
Rats weighed and given first injection (NaCl or LiCl). 10 minutes later, second injection (NaCl or NaB). 50 minutes later, overdosed with pentobarbital and transcardially perfused, first with 100 ml 0.15 M NaCl/NaNitrite/heparin, then 400ml of 4% PFA. Brain dissected, forebrain/brainstem blocked, post-fixed in 4% PFA for 1 h, then cryopotected in 30% sucrose for 2-3 days.
2022-2-26: DMU01-04,DMU9-12 perfused
2022-2-27: DMU05-8,DMU13-16 perfused
<br>
'''Brain Sectioning'''
36 x 40 micron sections through hindbrain (NTS)
2022-3-2: DMU01-04,DMU9-12 hindbrain sectioned
2022-3-3: DMU05-8,DMU13-16 hindbrain sectioned
'''Immunohistochemistry'''
anti-acetyl-Histone Primary: anti-acetyl-Histone H3 (Lys14) rabbit polyclonal, Millipore 07-353, Lot #3259085, r. 2019-09-26 (use at 1:2000)
Secondary: FITC conjugated Goat Anti-Rabbit, Jackson Labs 111-095-144, Lot #77909, r. 2008-05-24 (use at 1:100)
anti-c-Fos Primary: anti-c-Fos monoclonal antibody (E-8), Santa Cruz sc-166940, Lot # L1621, r. 2022-3-3 (use at 1:100)
Secondary: Texas Red conjugated Goat Anti-Mouse, Jackson Labs 115-075-146, Lot #72021, r. 2008-05-24 (use at 1:100)
[[Category:Taste Aversion]]
[[Category: CTA Experiments]]