Kobayashi K, Kobayashi H, Ueda M, Honda Y
Department of Ophthalmology and Visual Science, Kyoto University Graduate School of Medicine, Japan.
Exp Eye Res. 1997 May;64(5):719-26. doi: 10.1006/exer.1996.0262.
Retinal pigment epithelium (RPE) has essential roles that maintain functions of the neural retina. Previously, we described that the 3b5 monoclonal antibody (3b5 mAb) recognized the RPE cells in the chick embryonic eye. In the present study, a gene coding for the 3b5 antigen has been cloned and sequenced. A 10-day-old embryonic RPE cDNA library was constructed in lambda Uni-ZAP XR, and screened with the 3b5 mAb as a probe. The complete nucleotide sequence of the antigen recognized by the 3b5 mAb was obtained. The cDNA encodes a 735 amino acid protein with a calculated molecular mass of 80,147 Da. The deduced amino acid sequence shares 72% identity with the human 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) type IV. Northern blot analysis detected a 2.9 kb transcript. Immunohistochemical and in-situ hybridization studies revealed that 17 beta-HSD type IV was preferentially expressed in the RPE cells. The expression of steroid metabolizing enzymes in the RPE cells is able to create estrogenic environment, which may have a role of maintenance of the neural retina.
视网膜色素上皮(RPE)在维持神经视网膜功能方面起着至关重要的作用。此前,我们描述了3b5单克隆抗体(3b5 mAb)可识别鸡胚眼中的RPE细胞。在本研究中,编码3b5抗原的基因已被克隆和测序。构建了一个10日龄胚胎RPE的λUni-ZAP XR cDNA文库,并用3b5 mAb作为探针进行筛选。获得了3b5 mAb识别的抗原的完整核苷酸序列。该cDNA编码一个735个氨基酸的蛋白质,计算分子量为80147 Da。推导的氨基酸序列与人类IV型17β-羟基类固醇脱氢酶(17β-HSD)有72%的同一性。Northern印迹分析检测到一个2.9 kb的转录本。免疫组织化学和原位杂交研究表明,IV型17β-HSD在RPE细胞中优先表达。RPE细胞中类固醇代谢酶的表达能够创造雌激素环境,这可能对维持神经视网膜有作用。