Monoclonal antibody BM89 recognizes a novel cell surface glycoprotein of the L2/HNK-1 family in the developing mammalian nervous system.

A monoclonal antibody, BM89, obtained with Triton X-114-treated pig synaptic membranes as an immunogen, recognizes a neuronal antigen in the newborn porcine nervous system. By immunohistochemistry, BM89 staining was observed within the neuropil of all areas of the forebrain and spinal cord tested. In addition, BM89 labeled the cell bodies and proximal dendrites of spinal cord neurons. In the peripheral nervous system, BM89 immunoreactivity was present in a subpopulation of dorsal root ganglion neurons and was predominantly associated with non-myelinated axons in peripheral nerves. Initial biochemical characterization of the antigen in pig brain showed that it is an integral membrane glycoprotein with a molecular weight of 41,000. Moreover, it cross-reacts with the L2/HNK-1 carbohydrate epitope expressed by members of a large family of glycoproteins. Homologous antigens with molecular weights of 41,000-43,000 were identified in the rat, rabbit and fetal human brain. Immunoblotting and immunohistochemistry revealed that the epitope recognized by BM89 is developmentally regulated in the rat nervous system. In cryostat sections from rat cerebellum, spinal cord and dorsal root ganglia, an age-dependent decline of BM89 immunoreactivity was observed during postnatal development. In the cerebellum, the BM89 epitope was very abundant in cells of the external and the internal granular layers between postnatal days 5 and 15. During this period some staining was also identified in the developing molecular layer and the prospective white matter. Subsequently, and in the adult, overall staining was greatly reduced and remaining immunoreactivity was associated only with the internal granular layer. In the spinal cord and dorsal root ganglia, staining was very prominent at postnatal day 5; it decreased considerably thereafter and was barely detectable in the adult. Immunostaining of rat brain and dorsal root ganglion cultures revealed that the BM89 antigen is a cell surface molecule expressed by a subpopulation of central and peripheral nervous system neurons. The biochemical properties in conjunction with the topographical location and the developmental profile of the antigen recognized by BM89 suggest that it may represent a developmentally important recognition molecule.