Processing the spinal cord for in situ hybridization with radiolabelled oligonucleotides

Abstract

Publisher Summary The spinal cord receives sensory information from the trunk and limbs and is the final station for movement commands. Its anatomy and physiology are well defined and detailed knowledge about the segmental spinal-cord circuitry makes this structure ideal for experimental studies. As the first station to integrate somatosensory information in the central nervous system (CNS), the spinal cord is of outstanding interest to the field of pain research and therapy, and is extensively studied in models of traumatic injury or regeneration. This chapter describes the application of the in situ hybridization (ISH) method to investigate gene expression in the spinal cord of the adult rodent. This method has been used extensively to investigate the expression of neurotransmitters and to characterize neuroplastic changes following noxious sensory inputs in various experimental pain-model systems in the spinal cord. The ISH signal in the spinal cord can be quantified macroscopically from X-ray autoradiographs and in cellular resolution from emulsion-dipped sections. Quantification of silver-grain density in emulsion-dipped sections is a much more accurate, semiquantitative measure of the mRNA amount labeled because this quantification is selective for the ISH label over defined cells or structures.