GFA phospho-PDH and Taste Aversion
Examine levels of Phospho-Pyruvate Dehydrogenase (pPDH) by immunofluorescence in after intraoral conditioned taste aversion.
Based on Yang et al. 2024 PMID 38266644 and report by Roitman lab at SSIB 2024.
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.
For immunohistochemistry:
- for pPDH: Phospho-Pyruvate Dehydrogenase α1 (Ser293) (E4V9L) Rabbit mAb #37115 at 1:500 for immunofluoresence; BLUE secondary Jackson DyLight™ 405 AffiniPure™ Donkey Anti-Rabbit IgG (H+L) #711-475-152 [WHAT IS CONCENTRATION?]
- for c-Fos: mouse anti-c-Fos antibody (C-10; Santa Cruz) at a dilution of 1:1000. GET Antibody specification and secondary from Jason's work
phospho-PDH and CTA | |
---|---|
Taste Aversion | |
Expt Code | GFA |
Start Date | 2024-12-3 |
Investigators | Houpt, Cote, Trombley, Blakemore |
Funding | Chair's Fund |
Protocol | 202200000006 |
Expt Type | CTA |
Species | rat |
Drugs | lithium |
Data Link | https://houptlab.org/bartab/expt/?id=GFA |
Data
People
Jason Cote (breeding), Laura Blakemore and Paul Trombley (colony foundation and management)
CTA Conditioning
Agenda
Rats Arrive (December 3, 2024)
Rats (n=4 male, n=5 female, littermates, DOB: 7-???-2024) transferred from GFP-TH breeding protocol to individual cages.
Day 0: Surgery (December XXX, 2024)
Chronic Intraoral Catheters implanted under isoflurane anesthesia. All rats receive carprofen (XXX mg/kg) pre-operatively and 1 day post-op.
Days 1 - 3: Recovery (December XXXX - XXXX, 2024)
Weigh rats daily. Ad lib food and water. Carprofen (XXX mg/kg) on first day post-op.
Days 5-7: Habituation (December XXXX - XXXX, 2024)
Rats given intraoral infusion of 6 ml dH2O over 6 min to habituate to infusion procedure
Food removed on last day of habituation.
Noncontingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).
Day 8: Conditioning day 1 (December XXXX, 2024)
All rats given intraoral infusion of 6 ml sucrose 5% over 6 min.
Contingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).
Food returned overnight.
Day 9: Recovery Day (December XXXX, 2024)
Food removed.
Noncontingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).
Day 10: Conditioning day 2 (December XXXX, 2024)
All rats given intraoral infusion of 6 ml sucrose 5% over 6 min.
Contingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).
Food returned overnight.
Day 11: Recovery Day (December XXXX, 2024)
Food removed.
Noncontingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).
Day 12: Conditioning day 2 (December XXXX, 2024)
All rats given intraoral infusion of 6 ml sucrose 5% over 6 min.
Contingent group: Half of rats injected with LiCl (0.15M, 12 ml/kg).
Food returned overnight.
Day 13: Recovery Day (December XXXX, 2024)
Food removed.
Day 14: Expression Test (December XXXX, 2024)
All rats given intraoral infusion of 6 ml sucrose 5% over 6 min.
One hour after infusion, rats are overdosed with euthanasol and transcardially perfused. Brains postfixed for 1-2 hours, then stored in 0.1M PB.