Difference between revisions of "JCE Inhibitory DREADD Functional Validation"

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To test CNO as a uncondition stimulus for CTA in wild-type mice.
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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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 +
 
  
  
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| Name = Clozapine-N-Oxide Taste Aversion
 
| Name = Clozapine-N-Oxide Taste Aversion
 
| project = DREADDs and Magentic Field
 
| project = DREADDs and Magentic Field
| code = JCD
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| code = JCE
 
| protocol = 202200000006
 
| protocol = 202200000006
 
| funding = Chair's Fund
 
| funding = Chair's Fund
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| drugs = NaCl, CNO
 
| drugs = NaCl, CNO
 
| artifacts =  
 
| artifacts =  
| data = https://houptlab.org/bartab/expt/?id=JCD
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| data = https://houptlab.org/bartab/expt/?id=JCE
 
}}
 
}}
  
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==Data==  
 
==Data==  
  
'''[https://houptlab.org/bartab/expt/?id=JCD Raw Data Link]'''
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'''[https://houptlab.org/bartab/expt/?id=JCE Raw Data Link]'''
  
 
==People==  
 
==People==  
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==Agenda==
 
==Agenda==
  
Mice JAX C57BL/6J  Strain 000664 (n=8 male DOB +/- 3: 20Dec2022, n=8 female DOB +/- 3: 20Dec2022)
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Mice In House Colony FosCreER X hM4Di (n=11 male DOB +/-, n=12 female DOB +/-)
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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.
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 +
Pre Treatment (8/18/23) 2 weeks before starting water restriction
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4 groups:
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 +
1. Isotonic LiCl and 4-OHT, N=6
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 +
2. Isotonic LiCl and  sesame oil, N=6
  
Arrived 2023-2-6.
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3. Isotonic saline and 4-OHT, N=6
  
'''Day 0: Water removed''' (2023-6-19)
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4. Isotonic saline and sesame oil, N=5
  
  
'''Days 1 - 11: Water Restriction''' (2023-6-20 - 2023-6-30)
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'''Day 0: Water removed''' (2023-9-12)
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 +
 
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'''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.  
 
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.  
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In the afternoon (~4pm), mice will be given supplemental access to water for 30 min.
 
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 vehicle (10 ml/kg) or 1, 3 or 30 mg/kg  
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'''Day 12: Conditioning Day''' (2023-9-24)
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 +
Mice are given 10-min access to 0.125% saccharin solution. After 15 minutes mice are then injected with CNO (3mg/kg,10ml/kg, i.p) followed by LiCl (0.15M,20ml/kg, i.p) or isotonic saline after another 15 min.
  
  
 
Groups:  
 
Groups:  
 +
* LTL: Exp group (predicted to show attenuated taste aversion)
 +
* LVL: + Control (expect to form taste aversion)
 +
* VTV: - Control (expect to not form taste aversion)
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* VVV: - Control (expect to not form taste aversion)     
 +
  
* JCC01-04: Female, vehicle
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* JCE01-04: Female
* JCC05-08: Female, CNO
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* JCE05-08: Female
* JCC09-12: Male, vehicle
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* JCE09-12: Male
* JCC13-16: Male, CNO
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* JCE13-16: Male
  
 
After injection, ad lib water returned overnight.
 
After injection, ad lib water returned overnight.
  
'''Day 13 - 24:  Preference Tests''' (2023-3-17 - 2023-3-24 )
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'''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.
 
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.

Latest revision as of 18:43, 19 February 2024

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. After 15 minutes mice are then injected with CNO (3mg/kg,10ml/kg, i.p) followed by LiCl (0.15M,20ml/kg, i.p) or isotonic saline after another 15 min.


Groups:

  • LTL: Exp group (predicted to show attenuated 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)


  • 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.