Kalina M, Grimelius L
Department of Histology and Cell Biology, Sackler School of Medicine, Tel Aviv University, Israel.
J Histochem Cytochem. 1991 Feb;39(2):213-20. doi: 10.1177/39.2.1987265.
Rat alveolar Type II cells were immunostained with antibodies directed against chromogranin A (monoclonal, LK2H10) and chromogranins A and B (polyclonal, LKZM1U). The chromogranins or chromogranin-like proteins were identified in cells in lung tissue sections and isolated Type II cells at the light and electron microscopic levels. We used post-embedding immunoelectron microscopy, with immunogold, to detect the proteins' immunoreactivity in osmicated tissues. Gold particles were distributed over the phospholipid lamellae within the lamellar bodies of alveolar Type II cells and over the lattice structure of tubular myelin. Quantitative analysis of gold labeling densities in the various cell compartments indicated that only the latter two structures were specifically labeled. Controls, which included pre-absorption of both anti-chromogranin antibodies with excess chromogranin A or with native surfactant, resulted in a greater than 60% decrease in gold labeling. A possible role of chromogranins or chromogranin-like proteins as Ca2+ binding proteins in alveolar Type II cells is discussed.
用针对嗜铬粒蛋白A(单克隆抗体,LK2H10)以及嗜铬粒蛋白A和B(多克隆抗体,LKZM1U)的抗体对大鼠肺泡II型细胞进行免疫染色。在肺组织切片细胞和分离出的II型细胞中,通过光学显微镜和电子显微镜鉴定出了嗜铬粒蛋白或嗜铬粒蛋白样蛋白。我们采用包埋后免疫电子显微镜技术,利用免疫金法检测经锇酸处理组织中这些蛋白的免疫反应性。金颗粒分布在肺泡II型细胞板层小体内的磷脂片层以及管状髓磷脂的晶格结构上。对不同细胞区室中金标记密度的定量分析表明,只有后两种结构被特异性标记。对照实验包括用过量的嗜铬粒蛋白A或天然表面活性剂预吸附两种抗嗜铬粒蛋白抗体,结果金标记减少了60%以上。本文讨论了嗜铬粒蛋白或嗜铬粒蛋白样蛋白作为肺泡II型细胞中Ca2+结合蛋白的可能作用。