Enzyme histochemistry of the retina at light activation.

光激活时视网膜的酶组织化学。

基本信息

批准号：
60480101
负责人：
SATITO T.
金额：
$ 4.29万
依托单位：
Jichi Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1987
项目状态：
已结题

项目摘要

The development of the rapid freeze substitution method enables us to see excellent morphology in electron microscopy. Therefore, the possibility of enzyme histochemistry of cyclic nucleotide metabolizing enzymes on the quick feeze substituted rat retina was studied.The ultrastructure of the freeze substituted samples was excellent in showing the fine parallel arrangement of the disk membranes in the rod outer segnt.In spite of a long substitution fixation of 3 days, using 0.5% glutraldehyde and 6% tannic acid in acetone, guanylate cyclase, phosphodiesterase, CTPst, and 5'-nucleotidase were demonstrably retained. With the contrast enhancemen on biogical membranes by the 0.5-4% glutaraldehyde and 6% tannic acid in aceton, the site of phosphodiesterase activity was clearly visualized on the disk membranes of rod outer segment sepecially at cytoplasmic side. The low phosphodiesterase activity was observed in the rod outer segment from the dark adapted retina showing as some disk membranes were positive but the others were not. All disk membranes were appleared positive and enhanced at light conditions.The present investigation reveals that the freeze substitution technique can preserve not only fine morphology but also enzyme activity in cells and tissues at cerain physiological conditions.

快速冻结替代方法的发展使我们能够在电子显微镜中看到出色的形态。因此，研究了环环核苷酸代谢酶在快速费用取代的大鼠视网膜上的酶组织化学的可能性。冻结取代样品的超微结构非常出色，显示出圆盘膜的精细平行排列在杆状上的圆盘膜的良好平行布置，并使用3天的hong sepration of nyr sepration of yny hyder spritation of hy hy spritation of hy hyder centration of hy hyder spritation of hyd glut n.5％gluts of Hyde n.5％gluts n.5％glute gllut of hyd glut。在丙酮中，明显地保留了瓜烷酸盐环化酶，磷酸二酯酶，CTPST和5'-核苷酸酶。由于0.5-4％戊二醛对生物膜上的对比增强剂和乙酸的6％单氨酸，磷酸二酯酶活性的位点在杆外段的圆盘膜上清楚地看到了，尤其是在细胞质方面。在黑暗改装视网膜的杆外段中观察到低磷酸二酯酶活性，因为有些椎间盘膜是阳性的，但其他磁盘却没有。在光条件下，所有磁盘膜均得到阳性并增强。目前的研究表明，冻结替代技术不仅可以保留细胞和组织中细胞和组织中酶活性的酶活性。