Mirshahi M, Nicolas C, Mirshahi A, Hecquet C, d'Hermies F, Faure J P, Agarwal M K
INSERM U-86, Centre Universitaire des Cordeliers, Paris, France.
Biochem Biophys Res Commun. 1996 Feb 6;219(1):150-6. doi: 10.1006/bbrc.1996.0197.
Immunoblotting with a polyclonal antibody, directed against the mineralocorticoid receptor protein purified from rat kidney in presence of the receptor-specific ligand RU 26752, labeled a single 98-102 kDa band in soluble extracts from bovine retina and from cultured bovine retinal pigment epithelial cells, identical to the receptor in several other tissues from the rat. The antibody also immunoprecipitated the receptor-3H-RU 26752 complex in bovine retinal extract. The growth of the isolated pigment epithelial cells was inhibited by RU 26752 and ZK91587, two ligands specific to the mineralocorticoid receptor. Successive passages in culture led to the disappearance of immunoreactivity in Western blots, concurrently with the refractoriness of the cells to growth inhibition by the two antagonists. On sections of the human eye, mineralocorticoid receptor-specific immunofluorescence was observed in retinal cone cells, pigment epithelium, epithelium of ciliary body, iris, cornea and lens. To our knowledge, this is the first ever demonstration of the mineralocorticoid receptor in ocular tissues.
使用一种多克隆抗体进行免疫印迹分析,该抗体针对在受体特异性配体RU 26752存在下从大鼠肾脏纯化的盐皮质激素受体蛋白,在牛视网膜和培养的牛视网膜色素上皮细胞的可溶性提取物中标记出一条单一的98 - 102 kDa条带,与大鼠其他几种组织中的受体相同。该抗体还在牛视网膜提取物中免疫沉淀了受体 - 3H - RU 26752复合物。盐皮质激素受体的两种特异性配体RU 26752和ZK91587抑制了分离的色素上皮细胞的生长。连续传代培养导致免疫印迹中的免疫反应性消失,同时细胞对这两种拮抗剂的生长抑制产生抗性。在人眼切片上,在视网膜锥细胞、色素上皮、睫状体上皮、虹膜、角膜和晶状体中观察到盐皮质激素受体特异性免疫荧光。据我们所知,这是首次在眼组织中证明盐皮质激素受体的存在。