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Protocol for dissection and culture of murine dorsal root ganglia neurons to study neuropeptide release.

STAR protocols (2021-02-23)
Caroline Perner, Caroline L Sokol
ABSTRACT

In this protocol, we provide step-by-step instructions for dissection and culture of primary murine dorsal root ganglia (DRG), which provide an opportunity to study the functional properties of peripheral sensory neurons in vitro. Further, we describe the analysis of neuropeptide release by ELISA as a possible downstream application. In addition, isolated DRGs can be used directly for immunofluorescence, flow cytometry, RNA sequencing or proteomic approaches, electrophysiology, and calcium imaging. For complete details on the use and execution of this protocol, please refer to Perner et al. (2020).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Hanks′ Balanced Salt solution, Modified, with sodium bicarbonate, without phenol red, calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Cytosine β-D-arabinofuranoside hydrochloride, crystalline
Sigma-Aldrich
Glial Cell Line-derived Neurotrophic Factor from rat, recombinant, expressed in baculovirus infected Sf21 cells, lyophilized powder, suitable for cell culture, ≥97% (SDS-PAGE)