GFA phospho-PDH and Taste Aversion

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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:


phospho-PDH and CTA
Taste Aversion
Expt CodeGFA
Start Date2024-12-3
InvestigatorsHoupt, Cote, Trombley, Blakemore
FundingChair's Fund
Protocol202200000006
Expt TypeCTA
Speciesrat
Drugslithium
Data Linkhttps://houptlab.org/bartab/expt/?id=GFA


Data

Raw Data Link


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.

Histology