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AN ULTRASTRUCTURAL STUDY OF GLOMERULAR PERMEABILITY USING CATALASE AND PEROXIDASE AS TRACER PROTEINS

机译:用过氧化氢酶和过氧化物酶作为示踪蛋白对肾小球通透性的超微结构研究

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摘要

Mice were injected intravenously with beef liver catalase (mol wt 240,000) and very small doses of horseradish peroxidase (mol wt 40,000) and the site of localization of these enzymes in the kidney was studied by ultrastructural cytochemistry. 1 min after injection, catalase was present in glomerular capillary lumina and there was minimal permeation of the basement membrane. After 5–180 min, staining of the basement membrane increased progressively but was usually less than that in capillary lumina. At all time intervals the inner (sub-endothelial) layer of the basement membrane contained more reaction product than the lamina densa and the outer (subepithelial) layer. Catalase permeated the entire thickness of the basement membrane and extended up to the slit pore but not beyond the level of the slit diaphragm and was not seen in the urinary space or tubular lumina. Horseradish peroxidase permeated the whole thickness of the basement membrane within 2 min after injection; however, gradients of staining from the inner to outer layers of the basement membrane were frequently seen. The findings with both enzymes indicate that (a) the basement membrane restricts the passage of proteins over a wide range of molecular size with increasing impediment for larger molecules and (b) the slit pore functions as an additional barrier for molecules that cross the basement membrane.
机译:给小鼠静脉内注射牛肝过氧化氢酶(mol wt 240,000)和非常少量的辣根过氧化物酶(mol wt 40,000),并通过超微结构细胞化学研究这些酶在肾脏中的定位。注射后1分钟,过氧化氢酶存在于肾小球毛细血管腔内,并且基膜的渗透最小。 5–180分钟后,基膜的染色逐渐增加,但通常少于毛细管腔内的染色。在所有时间间隔,基膜的内(内皮下)层所含的反应产物都比椎板和外(胸膜上层)层多。过氧化氢酶贯穿基膜的整个厚度,并延伸至狭缝孔,但不超过狭缝隔膜的水平,并且在尿路空间或管状腔中均未见过。注射后2分钟内,辣根过氧化物酶渗入基膜的整个厚度。然而,从基底膜的内层到外层的染色梯度很常见。两种酶的发现均表明:(a)基底膜在较大分子大小范围内限制了蛋白质的通过,同时增大了对较大分子的阻碍,(b)狭缝孔为穿过基底膜的分子提供了额外的屏障。

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