Developmental stage-specific antigens in the nervous system of the cockroach.

A specific effort was made to obtain monoclonal antibodies that bind to macromolecules that play a role in the development of the nervous system. It was considered that good candidates for such molecules were those that were only transiently present in the embryonic nervous system. Hybridomas were prepared from spleen cells taken from mice that had been immunized with nerve cords from cockroach embryos at the 43-50% stage of development. The hybridoma supernatants were screened for antibody binding to frozen sections of both embryonic and adult thoracic ganglia. Cell lines that produced monoclonal antibodies that transiently bound to the embryonic nervous system were saved and cloned. These developmental stage-specific monoclonal antibodies either did not bind to the adult nervous system or bound to it with a pattern very different from that in the embryonic nervous system. The developmental stage-specific antigens detected by these monoclonal antibodies were organized into four categories based on the part of the embryonic nervous system in which they were transiently localized. These include binding to the cell bodies of all neurons, cell bodies of subsets of neurons or neuroblasts, subsets of axons, and the neuropile. Preliminary biochemical characterization of the antigens showed that many of these antibodies were recognizing carbohydrate epitopes. Functions for these antigens, most of which are components of the cell surface, are tentatively proposed.

Monoclonal antibodies to the cockroach nervous system.

In the cockroach nervous system individual motor neurons may be identified with respect to their position in the thoracic ganglia and to the muscles they innervate. When their axons are cut they have the ability to regrow such that when regeneration is completed they have specifically reinnervated their normal target muscles. This suggests the existence of a specific intercellular recognition process between motor neurons and muscles, and that neurons innervating different muscles are biochemically distinct from one another. The goal of this study was to use hybridoma techniques to obtain monoclonal antibodies that bind to some motor neurons and not others. Mice were injected with whole nerve cord and hybridoma supernatants were screened immunohistochemically on sections of ganglion and leg muscle. The monoclonal antibodies were categorized according to four types of specificity: tissue, regional, cell-type, and neuron-subset specificities. Antibodies expressing neuron-subset specificity were obtained very rarely. The probability of their occurrence could be increased by treating the mice with immunosuppressant drugs after initial administration of immunogen or by fixing the immunogen with paraformaldehyde in a manner similar to that of the tissue sections used in the screening process. Two of the neuron-subset specific monoclonal antibodies (MAbs) are of particular interest with respect to the goals of this study. They bind to axon terminals in the muscles of some neurons and not others. They do not bind to neuronal cell bodies in the ganglion, which makes identification of the neurons difficult. However, from the known innervation pattern of the coxal depressor muscles it appears that one of these MAbs selectively binds to axon terminals from either the inhibitory motor neurons or the dorsal unpaired median cells. Other antibodies of interest bind selectively to the synapse-rich neuropile in the ganglia or to peripheral parts of the nervous system like the nerve roots.