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晶状体代谢合作:一项利用冷冻置换放射自显影术和电子显微镜对小鼠晶状体转运和通透性进行可视化的研究。

Lens metabolic cooperation: a study of mouse lens transport and permeability visualized with freeze-substitution autoradiography and electron microscopy.

作者信息

Goodenough D A, Dick J S, Lyons J E

出版信息

J Cell Biol. 1980 Aug;86(2):576-89. doi: 10.1083/jcb.86.2.576.

Abstract

Transport of metabolites is demonstrated between compartments of the adult mouse lens by freeze-substitution autoradiography. In vivo patterns of lysine incorporation are compared with in vitro patterns of lysine, glucose, uridine, and deoxyglucose incorporation. Intracellular and extracellular distributions of tritiated metabolites are determined by comparison of transported substrates with the nontransported molecules of similar molecular size: mannitol and sucrose. The permeability of the lens intercellular spaces is probed with Procion Yellow at the level of fluorescence microscopy, and with horseradish peroxidase at the electron microscope level. Freeze-fracture electron microscopy reveals gap junctions between epithelial cells, between lens fibers, and between epithelial cells and lens fibers. Zonulae occludentes (tight junctions) are not routinely observed between epithelial cells in the mouse. This latter result is subject to species variation, however, since zonulae occludentes are abundant between chicken epithelial cells. The permeability results suggest that the lens cells are capable of metabolic cooperation, mediated by an extensive gap junction network.

摘要

通过冷冻置换放射自显影技术证明了成年小鼠晶状体各部分之间存在代谢物的转运。将赖氨酸掺入的体内模式与赖氨酸、葡萄糖、尿苷和脱氧葡萄糖掺入的体外模式进行比较。通过将转运的底物与分子大小相似的非转运分子(甘露醇和蔗糖)进行比较,确定氚标记代谢物的细胞内和细胞外分布。在荧光显微镜水平用普施安黄,在电子显微镜水平用辣根过氧化物酶探测晶状体细胞间隙的通透性。冷冻断裂电子显微镜显示上皮细胞之间、晶状体纤维之间以及上皮细胞与晶状体纤维之间存在间隙连接。在小鼠的上皮细胞之间通常未观察到紧密连接(闭锁小带)。然而,后一结果存在物种差异,因为在鸡的上皮细胞之间存在丰富的紧密连接。通透性结果表明,晶状体细胞能够通过广泛的间隙连接网络介导进行代谢协作。

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