Modeling Neuronal Injury in Mouse Primary Cerebellar Granule Neurons

In This Article

Abstract

Source: Laaper, M. et al., Modeling Neuronal Death and Degeneration in Mouse Primary Cerebellar Granule Neurons. J. Vis. Exp. (2017).

This video demonstrates the generation of a neuronal injury model in mouse primary cerebellar granule neurons. It outlines the steps involved in inducing neuronal injury using NMDA, an excitatory neurotransmitter receptor agonist, and glycine, a co-agonist. NMDA and glycine bind to specific neuronal receptors, causing their overstimulation and resulting in a massive intracellular calcium influx, which leads to neuronal cell damage and death.

Protocol

1. Experimental Preparation

NOTE: The following stock solutions can be prepared and maintained until use.

Dissection Solution
1. Dissolve 3.62 g of sodium chloride (NaCl), 0.2 g of potassium chloride (KCl), 0.069 g of sodium phosphate (NaH₂PO₄∙ H₂O), and 1.306 g of D-(+)-Glucose in 500 mL of distilled H₂O. Then, add 12.5 mL of HEPES Buffer, 450 µL of Phenol red solution, and 20 mL of Bovine serum albumin (BSA) fraction V to the solution. Adjust to pH 7.4 using 1 N sodium hydroxide (NaOH).
2. Sterilize with ....

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Results

Brain dissection process; A: incision direction diagram, B-C: brain tissue sample extraction.
Figure 1: Removal of mouse brain and dissection of cerebellum. (A) To extract the brain of a 6-7 day old mouse, using a pair of forceps, grasp the head and cut the skin anteriorly along the dotted lines using a pair of microdissection scissors.......