Bridges C C, El-Sherbeny A, Roon P, Ola M S, Kekuda R, Ganapathy V, Camero R S, Cameron P L, Smith S B
Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta, USA.
Histochem J. 2001 Mar;33(3):149-58. doi: 10.1023/a:1017991925821.
Caveolae are flask-shaped membrane invaginations present in most mammalian cells. They are distinguished by the presence of a striated coat composed of the protein, caveolin. Caveolae have been implicated in numerous cellular processes, including potocytosis in which caveolae are hypothesized to co-localize with folate receptor alpha and participate in folate uptake. Our laboratory has recently localized folate receptor alpha to the basolateral surface of the retinal pigment epithelium (RPE). It is present also in many other cells of the retina. In the present study, we asked whether caveolae were present in the RPE, and if so, whether their pattern of distribution was similar to folate receptor alpha. We also examined the distribution pattern of caveolin-1, which can be a marker of caveolae. Extensive electron microscopical analysis revealed caveolae associated with endothelial cells. However, none were detected in intact or cultured RPE. Laser scanning confocal microscopical analysis of intact RPE localized caveolin-1 to the apical and basal surfaces, a distribution unlike folate receptor alpha. Western analysis confirmed the presence of caveolin-1 in cultured RPE cells and laser scanning confocal microscopy localized the protein to the basal plasma membrane of the RPE, a distribution like that of folate receptor alpha. This distribution was confirmed by electron microscopic immunolocalization. The lack of caveolae in the RPE suggests that these structures may not be essential for folate internalization in the RPE.
小窝是存在于大多数哺乳动物细胞中的烧瓶状膜内陷结构。它们的特征是存在由小窝蛋白组成的条纹状衣被。小窝参与了众多细胞过程,包括小窝胞饮作用,据推测在该过程中小窝与α-叶酸受体共定位并参与叶酸摄取。我们实验室最近已将α-叶酸受体定位到视网膜色素上皮(RPE)的基底外侧表面。它也存在于视网膜的许多其他细胞中。在本研究中,我们探究了RPE中是否存在小窝,如果存在,其分布模式是否与α-叶酸受体相似。我们还检查了小窝蛋白-1的分布模式,它可以作为小窝的标志物。广泛的电子显微镜分析显示小窝与内皮细胞相关。然而,在完整的或培养的RPE中均未检测到小窝。对完整RPE进行激光扫描共聚焦显微镜分析,将小窝蛋白-1定位到顶端和基底表面,这种分布与α-叶酸受体不同。蛋白质印迹分析证实培养的RPE细胞中存在小窝蛋白-1,激光扫描共聚焦显微镜将该蛋白定位到RPE的基底质膜,这种分布与α-叶酸受体的分布相似。这种分布通过电子显微镜免疫定位得到证实。RPE中缺乏小窝表明这些结构可能对于RPE中的叶酸内化不是必需的。