Difference between revisions of "JCF Excitatory DREADD Functional Validation"
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| Line 5: | Line 5: | ||
CNO ordering information / lot numbers | CNO ordering information / lot numbers | ||
| − | CNO preparation | + | CNO preparation: Dissolved in isotonic saline 0.3mg/ml shaking |
| + | |||
| + | 4-OHT preparation: Dissolved in pharmaceutical grade sesame oil 5mg/ml shaking o/n 55c | ||
{{Infobox experiment|align=right | {{Infobox experiment|align=right | ||
| Name = Clozapine-N-Oxide Taste Aversion | | Name = Clozapine-N-Oxide Taste Aversion | ||
| project = DREADDs and Magentic Field | | project = DREADDs and Magentic Field | ||
| − | | code = | + | | code = JCF |
| protocol = 202200000006 | | protocol = 202200000006 | ||
| funding = Chair's Fund | | funding = Chair's Fund | ||
| investigators = Cote, Houpt | | investigators = Cote, Houpt | ||
| species = mice | | species = mice | ||
| − | | date = | + | | date = 2024-1-28 |
| type = CTA | | type = CTA | ||
| drugs = NaCl, CNO | | drugs = NaCl, CNO | ||
| artifacts = | | artifacts = | ||
| − | | data = https://houptlab.org/bartab/expt/?id=JCF} | + | | data = https://houptlab.org/bartab/expt/?id=JCF}} |
| Line 33: | Line 35: | ||
==Agenda== | ==Agenda== | ||
| − | Mice In House Colony FosCreER X | + | Mice In House Colony FosCreER X hM3Dq (n=7 male, n=16 female) |
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 ( | + | Pre Treatment (1/28/24) 2 weeks before starting water restriction |
4 groups: | 4 groups: | ||
| − | 1. Isotonic LiCl and 4-OHT, N= | + | 1. Isotonic LiCl and 4-OHT, N=12: Excitatory DREADDs are expected to be expressed |
| − | |||
| − | |||
| − | |||
| − | |||
| − | + | 2. Isotonic Saline and sesame oil, N=11: No DREADDs are expected to be expressed | |
| − | '''Day 0: Water removed''' ( | + | '''Day 0: Water removed''' (**2024-7-19) |
| Line 62: | Line 60: | ||
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''' ( | + | '''Day 12: Conditioning Day''' (2024-2-28) |
| − | Mice are given 10-min access to 0.125% saccharin solution, then injected with CNO (3mg/kg,10ml/kg, i.p) | + | Mice are given 10-min access to 0.125% saccharin solution, then injected with CNO (3mg/kg,10ml/kg, i.p) after 15 min |
Groups: | Groups: | ||
| − | * | + | * LTC: Exp group (Predicted to form taste aversion) |
| − | + | * VVC: Control (expect to not form taste aversion) | |
| − | |||
| − | * | ||
| − | * | + | * JCF01-04: ??? |
| − | * | + | * JCF05-08: ??? |
| − | * | + | * JCF09-12: ??? |
| − | * | + | * JCF13-16: ??? |
After injection, ad lib water returned overnight. | After injection, ad lib water returned overnight. | ||
Latest revision as of 15:38, 19 July 2024
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: Dissolved in isotonic saline 0.3mg/ml shaking
4-OHT preparation: Dissolved in pharmaceutical grade sesame oil 5mg/ml shaking o/n 55c
| Clozapine-N-Oxide Taste Aversion | |
|---|---|
| DREADDs and Magentic Field | |
| Expt Code | JCF |
| Start Date | 2024-1-28 |
| 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
People
Jason Cote
Agenda
Mice In House Colony FosCreER X hM3Dq (n=7 male, n=16 female)
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 (1/28/24) 2 weeks before starting water restriction 4 groups:
1. Isotonic LiCl and 4-OHT, N=12: Excitatory DREADDs are expected to be expressed
2. Isotonic Saline and sesame oil, N=11: No DREADDs are expected to be expressed
Day 0: Water removed (**2024-7-19)
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 (2024-2-28)
Mice are given 10-min access to 0.125% saccharin solution, then injected with CNO (3mg/kg,10ml/kg, i.p) after 15 min
Groups:
- LTC: Exp group (Predicted to form taste aversion)
- VVC: 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.