|Budget Amount *help
¥2,000,000 (Direct Cost : ¥2,000,000)
Fiscal Year 1994 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 1993 : ¥1,400,000 (Direct Cost : ¥1,400,000)
To reveal roles of differentiated microtubules in neurite outgrowth, we (1) studied significance of posttranslational modification in tubulin during development of the rat brain, and (2) tried to produce monoclonal antibodies inducing nerite outgrowth in the PC12 cell. For the first project, first, we established isoelectric focusing (IEF) , combined with immunoblot procedure using isotype-specific and modification site-specific anti-tubulin monoclonal antibodies, for analyzing posttranslational modification of tubulin. Using this IEF system, then, we found that modifications of alpha-and beta-tubulins were closely related to extension of neurites and maturation of dendrites, respectively. For the second project, first, we developed a living cell immunization procedure to produce monoclonal antibodies recognizing cell surface molecules on the living cells. Then, we found that one of anti-cell surface antibodies induced neurite outgrowth in the PC12 cell and that neuron-specific cytoskeletal proteins, such as neurofilament proteins NF-L and NF-M,and betaIII-tubulin, were expressed during the antibody-induced neurite outgrowth. Finally, we revealed that the antibody recognized a GPI-anchored molecule.