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Induction of c-Fos in rat brain by acute injections of lithium orotate (LiOR), LiCl, or NaCl.
{{Infobox experiment|align=right
| Name = Lithium Orotate c-Fos
| project = Taste Aversion
| code = SRA
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Rashid, Houpt
| species = rat
| date = 2026-10-15
| type = CTA
| drugs = lithium orotate, lithium chloride
| data =
}}
== People ==
Sheikh Rashid
== Agenda ==
=== Prepare Solutions ===
all injections made isotonic with TrisAmino:
* 0.3 M TrisAmino
36.34g / liter, autoclave, pH to 7.4 with 38% HCl
* 0.15 M LiOH
3.6g / liter (no need to autoclave)
* 0.15 M LiCl
6.45g / liter, autoclave
* LiOr = 75mM LiOrotate, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiOH, 0.47g orotic acid, pH to 7.4 with LiOH, filter
* LiCl = 75mM LiCl, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiCl, pH to 7.4 with LiOH, filter
* NaCl = 75mM NaCl, 150 mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M NaCl, pH to 7.4 with NaOH, filter
Perfusion solutions:
pre-perfusion saline
4% paraformaldehyde
=== Perfusion Day ===
Rats injected with one of the following solutions (staggered in pairs by 15-20min):
* 75 mM LiOr/Tris, 6 ml/kg ip. (SRA01, SRA04, SRA07)
* 75 mM LiCl/Tris, 6 ml/kg ip. (SRA02, SRA05, SRA08)
* 75 mM NaCl/Tris, 6 ml/kg ip. (SRA03, SRA06, SRA09)
These are same doses used in [[SNB Lithium Orotate CTA 2]], which showed that LiOr had greater potency in CTA conditioning than LiCl.
One hour after injection, rats are overdosed with sodium pentobarbital and transcardially perfused. Brains are dissected, and placed into 0.1M PB for storage, then transferred to 30% sucrose overnight before sectioning.
* 0: inject SRA01,SRA02
* 20: inject SRA03, SRA04
* 40: inject SRA05, SRA06
* 60: inject SRA07, SRA08 ; perfuse SRA01, SRA02
* 80: inject SRA09 ; perfuse SRA03, SRA04
* 100: perfuse SRA05, SRA06
* 120: perfuse SRA07, SRA08
* 140: perfuse SRA09
=== c-Fos Immunohistochemistry ===
Day 1: section forebrain, start immuno
Day 2: finish immuno
Day 3: mount tissue sections
Repeat for hindbrain...
=== Analysis ===
Digitize sections on microscope to count cells (a couple of days)