A video protocol for rapid dissection of mouse dorsal root ganglia from defined spinal levels

Abstract

Objective: Dorsal root ganglia (DRG) are heterogeneous assemblies of assorted sensory neuron cell bodies found in bilateral pairs at every level of the spinal column. Pseudounipolar afferent neurons convert external stimuli from the environment into electrical signals that are retrogradely transmitted to the spinal cord dorsal horn. To do this, they extend single axons from their DRG-resident somas that then bifurcate and project both centrally and distally. DRG can be dissected from mice at embryonic stages and any age post-natally, and have been extensively used to study sensory neuron development and function, response to injury, and pathological processes in acquired and genetic diseases. We have previously published a step-by-step dissection method for the rapid isolation of post-natal mouse DRG. Here, the objective is to extend the protocol by providing training videos that showcase the dissection in fine detail and permit the extraction of ganglia from defined spinal levels. Results: By following this method, the reader will be able to swiftly and accurately isolate specific lumbar, thoracic, and cervical DRG from mice. Dissected ganglia can then be used for RNA/protein analyses, subjected to immunohistochemical examination, and cultured as explants or dissociated primary neurons, for in-depth investigations of sensory neuron biology.

Altered Sensory Neuron Development in CMT2D Mice Is Site-Specific and Linked to Increased GlyRS Levels

“…DRG were extracted from either non-perfused or saline-perfused mice as previously described (Sleigh et al, 2016 , 2020b ). The most caudal pair of floating ribs and the large size of lumbar level 4 (L4) DRG and associated axon bundles were used as markers to consistently and accurately define the spinal level.…”

Editorial: Peripheral nerve anatomy in health and disease

“…Mitochondria were smaller and transported more quickly in sensory projections than in supporting glial cells. Primary DRG cultures from horses will be a useful tool for studying the impact of several, prevalent equine peripheral nerve diseases (Cahalan, Boehm, et al, 2022), and can be compared with cultures from chickens (Powell et al, 2014), mice (Sleigh, West, & Schiavo, 2020) and dogs (Ganchingco et al, 2019), among others, to determine whether there are differences in basic cellular processes (e.g., axonal transport) between species.…”