JCF Excitatory DREADD Functional Validation

To validate FosCreER x DREADD mice by testing if active cellular populations after LiCl exposure can be reactivated with CNO administration to induce a 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 Code	JCE
Start Date	2023-6-19
Investigators	Cote, Houpt
Funding	Chair's Fund
Protocol	202200000006
Expt Type	CTA
Species	mice
Drugs	NaCl, CNO
Data Link	https://houptlab.org/bartab/expt/?id=JCF

Data

Raw Data Link

People

Jason Cote

Agenda

Mice In House Colony FosCreER X hM3Dq (n=?? male DOB +/-, n=?? 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 (1/27/24) 2 weeks before starting water restriction 4 groups:

1. Isotonic LiCl and 4-OHT, N=6: Excitatory DREADDs are expected to be expressed

2. Isotonic LiCl and sesame oil, N=6: No DREADDs are expected to be expressed

3. Isotonic saline and 4-OHT, N=6: Due to the presence of 4-OHT but no LiCl, some DREADD expression is expected

4. Isotonic saline and sesame oil, N=5 No DREADD expression expected

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). ***Only LVL group will receive LiCl (0.15M,20ml/kg, i.p) after 10 min

Groups:

• LTL: Exp group (Predicted to form taste aversion)
• LVL: + Control (expect to form taste aversion)
• VTV: - Control (expect to not form taste aversion)
• VVV: - Control (expect to not form taste aversion)

• JCF01-04: ???
• JCF05-08: ???
• JCF09-12: ???
• JCF13-16: ???

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.