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.

A Selective Adenylyl Cyclase 1 Inhibitor Relieves Pain Without Causing Tolerance

“…Mice were humanely euthanized via decapitation under isoflurane anesthesia (open drop method) 90 min after the injection. DRGs (approximately C5 to L3) were dissected as previously described ( Sleigh et al, 2020 ), frozen in liquid N 2 , and stored in a −80°C freezer until used. On assay day, DRGs from a ST034307-injected mouse and its vehicle-matched control were thawed on ice.…”

Tozasertib Attenuates Neuropathic Pain by Interfering with Aurora Kinase and KIF11 Mediated Nociception

“…While cutting deep into abdominal musculature a caution was made not to cut spinal nerves. The muscular, fat layer, vertebras connected to L4, L5, L6, and spinal cord were removed and cleaned . DRGs, sciatic nerve, and spinal cord were identified and isolated and were placed in PBS followed by immediate transfer of all the isolated tissues to −80 °C.…”

“…The muscular, fat layer, vertebras connected to L4, L5, L6, and spinal cord were removed and cleaned. 41 DRGs, sciatic nerve, and spinal cord were identified and isolated and were placed in PBS followed by immediate transfer of all the isolated tissues to −80 °C. Tissue lysate was prepared by using RIPA buffer.…”

Chondroitin and glucosamine sulphate reduced proinflammatory molecules in the DRG and improved axonal function of injured sciatic nerve of rats

“…Five to ten hours after the last dose of treatment, the spinal column was dissected from the base of the skull to the level of the femurs, it was cut down the mid-line and the spinal cord extruded and meninges removed as described by Sleigh et al 60 , L3&L4 DRG were extracted and rinsed and stored in ice-cold phosphate buffer solution (PBS) and frozen for biochemical analysis. DRG was homogenized with the aid of a glass homogenizer at 4 °C in ice-cold saline (2 ml) (11 mmol L −1 Tris buffer, pH 7.4).…”