Validating Responsiveness of AAV-DIO-hM3Dq-DREADD
Katerina Rademacher, Ken Nakamura
Disclaimer
The protocols.io team notes that research involving animals and humans must be conducted according to internationally-accepted standards and should always have prior approval from an Institutional Ethics Committee or Board.
Abstract
To confirm responsiveness of AAV-DIO-hM3Dq-DREADD in DATIRESCre mice. Activation of DREADD receptors in dopamine neurons with CNO should induce a robust increase in locomotion.
Steps
Preparation
Mice should be at least two-weeks post-surgery.
Habituate mice to wireless running wheels in individually-housed cages.
See Victoria Vance, Katerina Rademacher, Ken Nakamura 2023. Behavior Tracking with Running Wheels. protocols.io https://protocols.io/view/behavior-tracking-with-running-wheels-cymrxu56
Prepare CNO for IP injection.
0.5mg/kg CNO dissolved in sterile saline, 500µL volume per mouse.
Prepared fresh the day of injection.
Procedure
Note: All injections should be performed at the same time of day, at least 2–3 hours before the dark cycle.
Day 1: IP inject mice with 500µL of sterile saline.
Day 2: IP inject mice with 500µL CNO.
Day 3 or 4: IP inject mice again with 500µL CNO.
Responders increase their running wheel activity in the 10-60min following IP injection of CNO. Exclude mice that do not respond to IP CNO from further analysis.
