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辣根过氧化物酶从脑脊液进入大鼠脉络丛的摄取,特别涉及跨上皮运输。

Uptake of horseradish peroxidase from CSF into the choroid plexus of the rat, with special reference to transepithelial transport.

作者信息

van Deurs B, Møller M, Amtorp O

出版信息

Cell Tissue Res. 1978 Feb 24;187(2):215-34. doi: 10.1007/BF00224366.

Abstract

Protein uptake from cerebral ventricles into the epithelium of the choroid plexus, and transport across the epithelium were studied ultrastructurally in rats. Horseradish peroxidase (HRP, MW 40,000) was used as protein tracer. Steady-state ventriculo-cisternal perfusion with subatmospheric pressure (-10cm of water) in the ventricular system was applied. HRP dissolved in artificial CSF was perfused from the lateral ventricles to cisterna magna for various times, and ventriculo-cisternal perfusion, vascular perfusion or immersion fixation with a formaldehyde-glutaraldehyde solution was performed. Coated micropinocytic vesicles containing HRP were seen both connected with the apical, lateral and basal epithelial surface and within the cells. Heavily HRP-labeled vesicles were often fused with the lining membrane of slightly labeled or unlabeled intercellular spaces. Since the apical tight junctions of the epithelium never appeared open or never contained HRP in the spaces between the fusion points, and since the intercellular spaces between adjacent epithelial cells below the junctions only infrequently contained tracer after 5 min, by increasing amounts after 15-60 min of HRP perfusion, a vesicular transport of HRP from the apical epithelial surface to the intercellular spaces, bypassing the tight junctions, is suggested. In addition to the transepithelial transport, micropinocytic vesicles also transported HRP to the lysosomal apparatus of the epithelial cells. With increasing length of exposure to HRP, a sequence of HRP-labeled structures could be evaluated, from slightly labeled apical vacuoles and multivesicular bodies to very heavily labeled dense bodies.

摘要

在大鼠中,利用超微结构研究了蛋白质从脑室进入脉络丛上皮以及跨上皮的转运过程。辣根过氧化物酶(HRP,分子量40,000)用作蛋白质示踪剂。在脑室系统中采用低于大气压(-10厘米水柱)的稳态脑室-脑池灌注。将溶解在人工脑脊液中的HRP从侧脑室灌注至小脑延髓池不同时间,然后进行脑室-脑池灌注、血管灌注或用甲醛-戊二醛溶液进行浸没固定。可见含有HRP的被膜微胞饮小泡既与上皮细胞的顶端、侧面和基底表面相连,也存在于细胞内。大量HRP标记的小泡常与轻度标记或未标记的细胞间隙的衬里膜融合。由于上皮细胞的顶端紧密连接从未出现开放状态,融合点之间的间隙中也从未含有HRP,并且由于在HRP灌注5分钟后,连接下方相邻上皮细胞之间的细胞间隙中很少含有示踪剂,而在15 - 60分钟后示踪剂含量逐渐增加,提示HRP通过小泡转运从上皮细胞顶端表面进入细胞间隙,绕过紧密连接。除了跨上皮转运外,微胞饮小泡还将HRP转运至上皮细胞的溶酶体装置。随着暴露于HRP时间的增加,可以评估一系列HRP标记的结构,从轻度标记的顶端空泡和多泡体到标记非常强烈的致密体。

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