JCE Inhibitory DREADD Functional Validation

Revision as of 17:31, 23 September 2023 by Jcote (talk | contribs) (→‎Agenda)

To validate FosCreER x DREADD mice by testing if active cellular populations after LiCl exposure can be inhibited with CNO administration to reduce/prevent taste aversion.



CNO ordering information / lot numbers

CNO preparation (e.g. vehicle, how its dissolved)

Clozapine-N-Oxide Taste Aversion
DREADDs and Magentic Field
Expt CodeJCE
Start Date2023-6-19
InvestigatorsCote, Houpt
FundingChair's Fund
Protocol202200000006
Expt TypeCTA
Speciesmice
DrugsNaCl, CNO
Data Linkhttps://houptlab.org/bartab/expt/?id=JCE


Data

Raw Data Link

People

Jason Cote

Agenda

Mice In House Colony FosCreER X hM4Di (n=11 male DOB +/-, n=12 female DOB +/-)

Drug Preparation: Isotonic LiCl (0.15M,20ml/kg, i.p) followed by 4-OHT prepared at 5mg/ml (50mg/kg,10ml/kg, i.p) or Sesame oil vehicle (10ml/kg i.p) after 10 min.

Pre Treatment (8/18/23) 2 weeks before starting water restriction 4 groups:

1. Isotonic LiCl and 4-OHT, N=6

2. Isotonic LiCl and sesame oil, N=6

3. Isotonic saline and 4-OHT, N=6

4. Isotonic saline and sesame oil, N=5


Day 0: Water removed (2023-9-12)


Days 1 - 11: Water Restriction (2023-9-13 - 2023-9-23)

Weigh mice daily. Ad lib food. Water access on consecutive days at noon: 3h, 3h, 3h, 1h, 1h, 1h, 30min, 30 min, 10 min, 10 min, 10 min.


Mice given water bottle and empty bottle, alternating position.

In the afternoon (~4pm), mice will be given supplemental access to water for 30 min.

Day 12: Conditioning Day (2023-9-24)

Mice are given 10-min access to 0.125% saccharin solution, then injected with CNO (3mg/kg,10ml/kg, i.p) followed by LiCl (0.15M,20ml/kg, i.p) or isotonic saline after 10 min


Groups: All mice receive CNO & LiCl injections

  • JCE01-04: Female
  • JCE05-08: Female
  • JCE09-12: Male
  • JCE13-16: Male

After injection, ad lib water returned overnight.

Day 13 - 24: Preference Tests (2023-9-25 - 2023-10-6 )

CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Mice were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 7 days.