Difference between revisions of "JCE Inhibitory DREADD Functional Validation"
| (6 intermediate revisions by the same user not shown) | |||
| Line 6: | Line 6: | ||
CNO ordering information / lot numbers | CNO ordering information / lot numbers | ||
| − | CNO preparation | + | CNO preparation: Dissolved in sterile isotonic saline 0.3mg/ml shaking |
{{Infobox experiment|align=right | {{Infobox experiment|align=right | ||
| Line 34: | Line 34: | ||
==Agenda== | ==Agenda== | ||
| − | Mice In House Colony FosCreER X hM4Di (n= | + | 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. | 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 | + | Pre Treatment (8/18/23) 2 weeks before starting water restriction |
4 groups: | 4 groups: | ||
| Line 47: | Line 47: | ||
3. Isotonic saline and 4-OHT, N=6 | 3. Isotonic saline and 4-OHT, N=6 | ||
| − | 4. Isotonic saline and sesame oil, N= | + | 4. Isotonic saline and sesame oil, N=5 |
| − | '''Day 0: Water removed''' (2023- | + | '''Day 0: Water removed''' (2023-9-12) |
| − | '''Days 1 - 11: Water Restriction''' (2023- | + | '''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. | ||
| Line 63: | Line 63: | ||
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- | + | '''Day 12: Conditioning Day''' (2023-9-24) |
| − | Mice are given 10-min access to 0.125% saccharin solution | + | 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) | ||
| + | * 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. | After injection, ad lib water returned overnight. | ||
| − | '''Day 13 - 24: Preference Tests''' (2023- | + | '''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 14:31, 19 July 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: Dissolved in sterile isotonic saline 0.3mg/ml shaking
| 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=JCE |
Data
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.