|Budget Amount *help
¥3,100,000 (Direct Cost : ¥3,100,000)
Fiscal Year 1998 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Fiscal Year 1997 : ¥2,000,000 (Direct Cost : ¥2,000,000)
1) Production of monoclonal antibodies against lamprey-retinal homogenate.
By this project, 23 new monoclonal antibodies were added to the series of antibodies we precedingly produced. According to their binding sites, the 23 antibodies were classified as follows : (1) Putative photopigment-specific antibodies including long photoreceptor cell (LPC) specific, Fl, Al and short (SPC) -specific, HI ; (2) LPC- and SPC -inner segment-specific, C12, H7, and F91 ; (3) Entire cell-body (LPC and SPC) -specific, G3, 04, and F92 ; (4) Several antibodies which were characterized by their binding to broad areas of glial processes, D2, D5, G3, D1, F11, A3, H6, D12, and A8 ; and (5) Pigment epithelium-specific, A1. In the present project, D2 was selected and investigated particularyin distal. D2 was found to bind with intermediate-sized filaments of Muller cell of the retina, supporting cells of the pineal and parapineal bodies, and ependymal cells bordering the ventricular system. By Western blotting against retinal homogenate, D2 antibody was enhanced by a remarkable band at 76kD.The nature of the protein is recognized by D2 antibody will be subject of the next project.
2) Electron microscopy of immunogold binding sites.
By freeze replica labeling method, a component of rhodopsin in the form dense IMP gathering more or less with immunogold conjugated H16 antibody. For this method, antibody H16 was most effective, so that it was useful as a standard for the EM immunocytochemistry. For D2, however, was detected on intermediate-sized filaments which were mostly hidden under shadowing deposition.