Glycobiological Study of Physiological Roles of Interphotoreceptor Matrix

光感受器间基质生理作用的糖生物学研究

基本信息

批准号：
09671809
负责人：
UEHARA Fumiyuki
金额：
$ 1.92万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671809/
关键词：
Retina Interphotoreceptor matrix Glycobiology Lectin Rat Cow Retinal adhesion Light-response 網膜移植 cDNA

项目摘要

(a) Study of experimental conditions for successful retinal transplantation : Immunohistochemical studies revealed that macrophages are present in the proliferating cells of the rat retina induced by constant fluorescence light exposure. Two lectins were purified from bovine retinas by affinity chromatography using a lactose column. We isolated the cDNAs of the lectins by screening bovine retinal cDNA libraly using the polyclonal antibodies against the lectins. The sequence analysis revealed that the 15kD- and 35kD-lectins correspond to galectin and unknown lectin, respectively. We will use these retinal lectins for the experiments of retinal transplantation instead of plant lectins.(b) Study of mechanism of retinal adhesion mediated by the O-type-glycoconjugates in the interphotoreceptor matrix : Immunohistochemical studies revealed that MUC2 is not present in the retina. This result suggests that MUC2 is not related to retinal adhesion. We isolated the, cDNA of a core protein of the O-type-glycoconjugate by screening bovine retinal cDNA libraly using polyclonal antibodies. The sequence analysis revealed that the core protein is rich in serine and threonine. We will use the fusion protein from expression of the cDNA for the study of retinal adhesion.(c) Study of light-response of the N-type-glycoconjugates in the interphotoreceptor matrix : The light- response was detected by use of a fixative containing glutaraldehyde, whereas it was not deteted using paraformaldehyde alone for the perfusion-fixation of rats in light- and ultra-microscopic sudies. These results suggest that the N-type-glycoconjugates may move in the subretinal space without bridging of the interphotoreceptor matrix.

(a) 成功视网膜移植的实验条件研究：免疫组织化学研究表明，在持续荧光照射诱导的大鼠视网膜增殖细胞中存在巨噬细胞。使用乳糖柱通过亲和层析从牛视网膜中纯化两种凝集素。我们通过使用针对凝集素的多克隆抗体筛选牛视网膜cDNA文库来分离凝集素的cDNA。序列分析表明15kD-和35kD-凝集素分别对应于半乳糖凝集素和未知凝集素。我们将使用这些视网膜凝集素代替植物凝集素进行视网膜移植实验。(b)光感受器间基质中O型糖缀合物介导的视网膜粘附机制的研究：免疫组织化学研究表明视网膜中不存在MUC2 。这一结果表明MUC2与视网膜粘连无关。我们通过使用多克隆抗体筛选牛视网膜 cDNA 文库，分离了 O 型糖缀合物核心蛋白的 cDNA。序列分析显示核心蛋白富含丝氨酸和苏氨酸。我们将使用 cDNA 表达的融合蛋白来研究视网膜粘附。(c) 光感受器间基质中 N 型糖缀合物的光响应研究：通过使用含有戊二醛，而在光显微镜和超显微镜研究中单独使用多聚甲醛对大鼠进行灌注固定时并未检测到它。这些结果表明，N 型糖缀合物可以在视网膜下间隙中移动，而不桥接感光器间基质。