Induction of Long-Term Depression in Cerebellar Purkinje Neurons in a Mouse Brain Slice

In This Article

Abstract

Source: Yamaguchi, K., et al. Assessment of Long-term Depression Induction in Adult Cerebellar Slices. J. Vis. Exp. (2019)

In this video, a mouse cerebellar brain slice was used to investigate synaptic interactions between parallel fibers, climbing fibers, and Purkinje cells, which are essential for motor control. Stimulation of climbing fibers induced long-term depression (LTD) in Purkinje cells, weakening synaptic strength and resulting in reduced response to parallel fiber stimulation.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Brain Slicing

Orient the sample such that the dorsal side of the cerebellum is on the front side. Pour ice-cold cutting ACSF (Artificial cerebrospinal fluid), containing 1 µM TTX(Tetrodotoxin), sufficient to immerse the cerebellum completely. Place a gas tube into the ACSF and start bubbling with the O₂/CO₂ gas mixture.
Remove the arachnoid mater using a fine tweezer under binoculars. Cut the cerebellar peduncle with a blade, and remove the brainstem....

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Results

Neuronal stimulation diagram; electrophysiological recording setups; depolarization waveforms.
Figure 1: Schematic illustration of protocols to induce LTD in PF-PC synapse. (A) Protocol 1 Cj. 1 PF and 1 CF stimuli are applied simultaneously 300 times at 1 Hz (5 min) under current-clamp conditions. Electrode for whole-cell recording con.......