Sacral Spinal Cord Transection and Isolated Sacral Cord Preparation to Study Chronic Spinal Cord Injury in Adult Mice.

Carmelo Bellardita,Ole Kiehn,Maite Marcantoni,Peter Löw

doi:10.21769/bioprotoc.2784

Abstract

Spinal cord injury (SCI) is characterized by multiple sensory/motor impairments that arise from different underlying neural mechanisms. Linking specific sensory/motor impairments to neural mechanism is limited by a lack of direct experimental access to these neural circuits. Here, we describe an experimental model which addresses this shortcoming. We generated a mouse model of chronic spinal cord injury that reliably reproduces spasticity observed after SCI, while at the same time allows study of motor impairments in vivo and in an in vitro preparation of the spinal cord. The model allows for the combination of mouse genetics in in vitro and in vivo conditions with advanced imaging, behavioral analysis, and detailed electrophysiology, techniques which are not easily applied in conventional SCI models.

Highlights

Note: All experiments should be performed in accordance with relevant guidelines and regulations
We describe the technical aspects of sacral spinal cord transection in adult mice, and the subsequent use of in vitro sacral spinal cord preparations for direct examination of the neural mechanisms which cause spasm in chronic spinal cord injury
The glass capillary is pulled, and the tip is manually adapted to the spinal cord

Summary

Materials and Reagents

Lidocaine Hydrochloride (Sigma-Aldrich, catalog number: BP214) 17. Sodium chloride (NaCl, Sigma-Aldrich, catalog number: 433209) Note: This product has been discontinued. Potassium chloride (KCl, Sigma-Aldrich, catalog number: P9541) 27. Sodium bicarbonate (NaHCO3, Sigma-Aldrich, catalog number: S5761) 29. Calcium chloride (CaCl2, Sigma-Aldrich, catalog number: 449709) 32. Magnesium chloride (MgCl2, Sigma-Aldrich, catalog number: V000149) 33. 1. Forceps (Fine Science Tools, catalog number: 11251-10) 2. Toothed forceps (Fine Science Tools, catalog number: 11154-10) 3. Dumont No 2 laminectomy forceps (Fine Science Tools, catalog number: 11223-20) 5. Fine Scissors–Tungsten Carbide (Fine Science Tools, catalog number: 14568-09) 6. Fine Scissors–Tungsten Carbide & ToughCut (Fine Science Tool, catalog number: 14558-11) 7. Scalpel (Fine Science Tools, catalog number: 10020-00) 9.

Procedure

Findings

Data analysis