Aiming at a phylogenetic tree of vertebrate photoreceptor cells on the basis of immunocytochemistry

1)抗七鳃鳗-视网膜匀浆的单抗的制备：通过该项目，在我们先前生产的一系列抗体的基础上增加了23个新的单抗。根据结合部位，23种抗体可分为：(1)可能的光色素抗体，包括长感光细胞(LPC)特异的F1、A1和短(SPC)特异的HI；(2)LPC和SPC内节段特异的C12、H7和F91；(3)整个细胞体(LPC和SPC)特异的G3、04和F92；(4)几种抗体，其特征是与广泛的神经胶质突起区域结合，D2、D5、G3、D1、F11、A3、H6、D12和A8；和(5)色素上皮特异的，A1。在本课题中，我们选择了D2，并对其进行了详细的远端研究。D2与视网膜Muller细胞、松果体和副松果体的支持细胞以及与脑室系统接壤的室管膜细胞的中等大小的细丝结合。通过免疫印迹和视网膜匀浆杂交，D2抗体在76kD处增强了一条显著的条带。D2抗体识别该蛋白的性质将成为下一步研究的对象。2)免疫金结合部位的电子显微镜观察。该方法以抗体H16效果最好，可作为EM免疫细胞化学的标准。然而，在中等大小的细丝上检测到了D2，这些细丝大多隐藏在阴影沉积下。

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.