Calvo M, Tebar F, Lopez-Iglesias C, Enrich C
Departament de Biologia Cellular, Institut d' Investigacions Biomèdiques August Pi I Sunyer, Facultat de Medicina, Universitat de Barcelona, Barcelona, Spain.
Hepatology. 2001 May;33(5):1259-69. doi: 10.1053/jhep.2001.23937.
Caveolae are small pits on the plasma membrane found in several, if not all, differentiated cells. They are involved in potocytosis, endocytosis, transcytosis, membrane trafficking, and signal transduction. Although caveolin has recently been identified in subcellular fractions from rat liver there is no clear-cut morphologic evidence for the presence of prototypical caveolae on the surface of hepatocytes. In this study the presence of caveolae at the cell surface of hepatocytes was directly shown by rapid-freeze, deep-etching electron microscopy. Moreover, combined deep-etching and immunogold techniques revealed caveolin in caveolae of the dorsal membrane of primary culture hepatocytes. Using reagents that perturb membrane cholesterol and interfere with endocytosis through the caveolae, a caveolae-dependent internalization of cholera toxin B and retinol-binding protein by hepatocytes in primary culture was shown. Finally, immunocytochemical analysis of caveolin in nonparenchymal cells of the rat liver showed its presence in Kupffer and stellate cells, however no caveolin could be detected in endothelial cells.
小窝是在多种(即便不是所有)分化细胞的质膜上发现的小凹陷。它们参与液相内吞、胞吞作用、转胞吞作用、膜运输和信号转导。尽管最近在大鼠肝脏的亚细胞组分中鉴定出了小窝蛋白,但在肝细胞表面尚无明确的典型小窝存在的形态学证据。在本研究中,通过快速冷冻、深度蚀刻电子显微镜直接显示了肝细胞表面存在小窝。此外,结合深度蚀刻和免疫金技术在原代培养肝细胞背膜的小窝中发现了小窝蛋白。使用扰乱膜胆固醇并干扰通过小窝的胞吞作用的试剂,显示原代培养的肝细胞通过小窝依赖性内化霍乱毒素B和视黄醇结合蛋白。最后,对大鼠肝脏非实质细胞中小窝蛋白的免疫细胞化学分析显示其在库普弗细胞和星状细胞中存在,然而在内皮细胞中未检测到小窝蛋白。