Difference between revisions of "JCE Inhibitory DREADD Functional Validation"

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To validate FosCreER x DREADD mice by testing if active cellular populations after LiCl exposure can be inhibited with CNO administration: reducing/preventing taste aversion.  
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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.  
  
  
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Mice In House Colony FosCreER X hM4Di (n=10 male DOB +/-, n=12 female DOB +/-)
 
Mice In House Colony FosCreER X hM4Di (n=10 male DOB +/-, n=12 female DOB +/-)
  
Drug Preparation: Isotonic LiCl (0.15M,20ml/kg, i.p), 4-OHT prepared at 5mg/ml (50mg/kg,10ml/kg, i.p), Sesame oil vehicle (10ml/kg i.p)
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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 2 weeks before starting water restriction, all injections are given concomitantly
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Pre Treatment 2 weeks before starting water restriction
 
4 groups:
 
4 groups:
  

Revision as of 13:57, 10 August 2023

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=10 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 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=4


Day 0: Water removed (2023-8-26)


Days 1 - 11: Water Restriction (2023-8-27 - 2023-9-06)

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 (2023-3-16)

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) after 10 min


Groups:

  • JCC01-04: Female, vehicle
  • JCC05-08: Female, CNO
  • JCC09-12: Male, vehicle
  • JCC13-16: Male, CNO

After injection, ad lib water returned overnight.

Day 13 - 24: Preference Tests (2023-3-17 - 2023-3-24 )

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.