Harvesting DRG from Rat Spine: A Procedure to Extract Dorsal Root Ganglion from the Spine of a Rat Model

Abstract

Source: Schmidt, L. B. et al. The Chick Chorioallantoic Membrane In Vivo Model to Assess Perineural Invasion in Head and Neck Cancer. J. Vis. Exp. (2019)

In this video, we demonstrate the extraction of dorsal root ganglion or DRG from a harvested rat spine for downstream applications.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Harvest and Preparation of DRGs (estimated timing: 2 h, day 8)

NOTE: Experiments with mice and rats require approval from the IACUC. In some countries, the use of chicken eggs also requires approval.

1. Get six to seven week-old Sprague Dawley rats (~200 g in weight) to extract DRGs.
   NOTE: One rat should yield ~40 cervical and thoracic DRGs. Mouse DRG also integrates in the CAM; however, the conditions....

Results

Figure 1: DRG extraction on day 8. A. Rat schematic illustrating the anatomical location of the spine. B. Diagr.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
PBS (1x) pH 7.4	Gibco	# 10010-023	Phosphate Buffered Saline
Sprague Dawley rats (females)	Charles River laboratories	Strain code: 001	6-7 weeks old (190-210g in weight)
Dumont # 5 fine forceps	Fine Science Tools (FST)	# 11254-20	Used to harvest DRG
Extra fine Graefe forceps, curved	Fine Science Tools (FST)	# 11151-10	Used to graft DRG onto the CAM on day 8 and to harvest CAM tissue on day 17
Extra fine Graefe forceps, straight	Fine Science Tools (FST)	# 11150-10	Used to graft DRG onto the CAM on day 8 and to harvest CAM tissue on day 17
DMEM (1x)	Gibco	# 11965-092	Dulbeecco`s Modified Eagle Medium
CellTracker Red CMTPX fluorescent dye	Life Technologies	# C34552	Reconstitute 50µg in 40µL of DMSO and stock at -20oC. Use 1µL of stock solution/mL of culture medium
Pen/Strep	Gibco	# 15140-122	10,000 Units/mL Penicilin, 10,000 µg/mL Streptomycin
HI FBS	Gibco	# 10082-147	Heat-inactivated Fetal Bovine Serum
DMSO	Fisher Bioreagents	# BP231-100	Dimethyl Sulfoxide