Glycine neurons in the brain and spinal cord. Antibody production and immunocytochemical localization

Abstract

Antibodies were raised against glycine and they were specific for immunocytochemistry. Obtained from rabbits immunized with glycine conjugated to glutaryled protein-carriers, antisera were then purified by adsorption on the various glutaraldehyde-conjugated protein-carriers. Using a modified ELISA method, their specificity was determined in competition experiments between conjugated glycine and either non-conjugated glycine or other conjugated amino acids or derivatives, preincubated with anti-glycine antibodies. Calculated at half-displacement, the resulting cross-reactivity ratios showed conjugated glycine to be the best recognized compound. By revealing the presence of the majority of the glycine-containing cell bodies in the brainstem and spinal cord, immunocytochemical applications of glycine antibodies confirmed their use as specific tools for a better understanding of the role of glycine in the central nervous system.