van Kuppevelt T H, Cremers F P, Domen J G, van Beuningen H M, van den Brule A J, Kuyper C M
Eur J Cell Biol. 1985 Jan;36(1):74-80.
In order to localize and characterize proteoglycans in human lung alveoli, we have used the cationic dye Cuprolinic Blue according to the critical electrolyte concentration method. After staining, five types of Cuprolinic Blue-positive filaments become apparent: two types in the basement membranes of type I and type II epithelial cells respectively and lying in one or two layers; one type, more scattered, localized in the basement membrane of the endothelial cells and another kind associated with collagen fibrils and separated from each other according to the main banding period (+/- 60 nm) of these fibrils. Finally, there was a type of filament which was only locally present at a variety of places. The basement membrane filaments were sensitive to heparinase, heparitinase, pronase (without prefixation) and nitrous acid treatment, but not to Streptomyces hyaluronidase, neuraminidase, chondroitinase ABC, chondroitinase AC, pronase (after prefixation) and 2.0 M MgCl2 treatment. The basement membrane filaments, therefore, represent heparan sulphate-containing proteoglycans. On the other hand, the collagen fibril associated filaments were sensitive to treatment with heparinase, chondroitinase ABC and pronase (without prefixation), but insensitive to Streptomyces hyaluronidase, neuraminidase, nitrous acid, heparitinase, chondroitinase AC, pronase (after prefixation) and 2.0 M MgCl2 (after prefixation) treatment. These filaments thus represent iduronic acid-rich dermatan sulphate-containing proteoglycans. Several physiological functions for these proteoglycans are discussed.
为了定位和鉴定人肺肺泡中的蛋白聚糖,我们根据临界电解质浓度法使用了阳离子染料铜蓝。染色后,出现了五种铜蓝阳性细丝:分别位于I型和II型上皮细胞基底膜中的两种,呈一层或两层排列;一种分布更分散,位于内皮细胞基底膜中;另一种与胶原纤维相关,并根据这些纤维的主带周期(±60nm)彼此分开。最后,有一种细丝仅局部存在于多个部位。基底膜细丝对肝素酶、硫酸乙酰肝素酶、链霉蛋白酶(未预固定)和亚硝酸处理敏感,但对透明质酸酶、神经氨酸酶、软骨素酶ABC、软骨素酶AC、链霉蛋白酶(预固定后)和2.0M氯化镁处理不敏感。因此,基底膜细丝代表含硫酸乙酰肝素的蛋白聚糖。另一方面,与胶原纤维相关的细丝对肝素酶、软骨素酶ABC和链霉蛋白酶(未预固定)处理敏感,但对透明质酸酶、神经氨酸酶、亚硝酸、硫酸乙酰肝素酶、软骨素酶AC、链霉蛋白酶(预固定后)和2.0M氯化镁(预固定后)处理不敏感。因此,这些细丝代表富含艾杜糖醛酸的含硫酸皮肤素的蛋白聚糖。文中讨论了这些蛋白聚糖的几种生理功能。