Simionescu M, Simionescu N, Palade G E
J Cell Biol. 1982 Nov;95(2 Pt 1):425-34. doi: 10.1083/jcb.95.2.425.
Cationized ferritin (CF) was injected interstitially to study the distribution of anionic sites on the basement membrane and abluminal aspect of the endothelium in the fenestrated capillaries of the mouse pancreas and intestinal mucosa. Extensive, but uneven removal of the basement membrane was obtained by collagenase perfusion of the vasculature before CF labeling. In the absence of collagenase treatment, CF label was essentially restricted to the lamina rara externa of the basement membrane and occurred in clusters distributed in a relatively ordered planar lattice. After collagenase digestion, labeling of the lamina rara interna and of the abluminal aspect of the endothelium became possible. In the lamina rara interna, the CF label occurred in clusters with a distribution comparable to that found in the lamina rara externa. On the abluminal aspect of the endothelium, the plasmalemma proper was extensively, though variably, labeled. Coated pits were heavily labeled, whereas the membranes and stomatal diaphragms of plasmalemmal vesicles and transendothelial channels remained free of CF decoration. In contradistinction with the heavy labeling of their luminal aspects, the abluminal surface of the fenestral diaphragms were free of any CF decoration. Pronase treatment removed all anionic sites detectable by CF binding. The findings establish the existence of differentiated microdomains on the abluminal aspect of the endothelial plasmalemma and suggest that the capillary wall selects permeant macromolecules according to charge, in addition to size.
注射阳离子铁蛋白(CF)以研究小鼠胰腺和肠黏膜有孔毛细血管中基底膜和内皮细胞腔外侧面阴离子位点的分布。在进行CF标记之前,通过对脉管系统进行胶原酶灌注,可广泛但不均匀地去除基底膜。在未进行胶原酶处理的情况下,CF标记基本上局限于基底膜的外疏松层,呈簇状分布,排列相对有序。胶原酶消化后,可对内疏松层和内皮细胞腔外侧面进行标记。在内疏松层,CF标记呈簇状分布,与外疏松层相似。在内皮细胞腔外侧面,质膜本身被广泛标记,但标记程度不一。有被小窝被大量标记,而质膜小泡和跨内皮通道的膜及气孔隔膜未被CF标记。与它们的腔面被大量标记形成对比的是,窗孔隔膜的腔外表面没有任何CF标记。链霉蛋白酶处理可去除所有能被CF结合检测到的阴离子位点。这些发现证实了内皮细胞质膜腔外侧面存在分化的微结构域,并表明毛细血管壁除了根据大小选择通透大分子外,还根据电荷进行选择。