Nov 12, 2024
Open-field experiments from Alcacer et al 2024
- 1Alcala University
Protocol Citation: Cristina Alcacer 2024. Open-field experiments from Alcacer et al 2024. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr9e9bvmk/v1
Manuscript citation:
Abnormal hyperactivity of specific striatal ensembles encodes distinct dyskinetic behaviors revealed by high-resolution clustering
Cristina Alcacer, Andreas Klaus, Marcelo Mendonça, Sara F. Abalde, Maria Angela Cenci, Rui M. Costa
bioRxiv 2024.09.06.611664; doi: https://doi.org/10.1101/2024.09.06.611664
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 01, 2024
Last Modified: November 12, 2024
Protocol Integer ID: 111382
Keywords: ASAPCRN
Abstract
Open field experimental set up and protocol for Alcacer et al 2024.
Guidelines
The inertial sensor should secured to a connector or holder placed on the back of the microendoscope with a consistent alignment of its axes. If there is a different alignment and axes used, there may need to be corrections made in pre-processing before subsequent data analyses.
Materials
-white open-field arena (40 cm X 40 cm) with transparent acrylic bottom and side placed inside a sound-attenuating chamber.
-Video camera (Flea3, Point Grey Research) placed at the bottom of the arena recording at 30-40 frames per second (fps)
-Wireless inertial measurement unit (IMU), Champalimaud Hardware 22 Platform; WEAR motion sensor system - https://www.cf-hw.org/harp/wear)
-Microendoscope (Inscopix)
Head-mount Habituation
Head-mount Habituation
Before starting experiments, habituate each mouse for 1 hour per day for 2-3 days to the head-mount equipment.
Mouse is lightly anesthetized with isoflurane to facilitate mounting of the equipment.
Attach a replica of the microendoscope that is the same weight as the real one (2g) and the actual wireless inertial sensor (1.8g).
Allow mouse to wake from the anesthesia.
Monitor mouse while it habituates to the equipment for 1 hour.
Lightly anesthetized with isoflurane to facilitate removing the equipment.
Recover mouse in its home cage and repeat for a total of 2-3 days.
Experimental day
Experimental day
Mouse is lightly anesthetized with isoflurane to facilitate mounting of the equipment.
Attach the microendoscope (2g) and the wireless inertial sensor (1.8g).
Place mouse in the open field box.
Wait 15 minutes after mouse has recovered from the anesthesia before starting the experiment/recording.
Begin simultaneous video recording, striatal activity recording and acceleration monitoring. Note: the microendoscope and IMU are left in place all the session, that is, during the baseline recording, the 20 min waiting period and the 10 min recording after LD/VEH.
Baseline recording was taken for 10 minutes.
Remove mouse from open field arena and inject i.p. with L-DOPA or vehicle (see doi: xxx).
Place mouse in a cage next to the arena and allow for a 20 minute waiting period for treatment to take effect.
Place mouse back in the open field arena and record/image for another 10 minutes.
Stop recording and remove mouse from open field arena.
Lightly anesthetized with isoflurane to facilitate removing the equipment.
Recover mouse in its home cage and repeat for up to 3-4 days.