The investigator has generated four monoclonal antibodies that bind to basement membranes in peripheral nerve and cardiac and skeletal muscle, but do not recognize any epitopes in the central nervous system. These antibodies interfere with neurite growth in vitro. The location of these determinants and their functional activity suggest that these antibodies recognize constituents important for allowing successful axonal regeneration in the peripheral nervous system. The PC1C5 antigen is of particular interest since this antibody does not bind to the Schwann cell basement membrane in sections of sciatic nerves from homozygous dystrophic mice (dy/dy); all other antibodies tested (34 in total) do stain Schwann cell basement membranes in dy/dy mice. The objectives of the research are to: 1) test the PC1C5 antigen for its ability to promote neurite outgrowth; 2) obtain partial amino acid sequence for this antigen and look for homology to other previously characterized proteins; 3) screen cDNA expression libraries with the four antibodies or with synthetic oligonucleotide probes based on the sequences obtained in objective 2; 4) compare the molecular differences between PC1C5 antigens synthesized by Schwann cells and muscle cells; 5) screen libraries from various normal and dy/dy tissues with probes obtained from expression libraries in objective 3; and 6) generate Schwann cell lines from normal and dy/dy mice in an attempt to provide an in vitro model for the dystrophic mutation.
