Li Y C, Hayes S, Young A P
Neurobiotechnology Center, Ohio State University, Columbus 43210, USA.
J Mol Neurosci. 1997 Apr;8(2):145-58. doi: 10.1007/BF02736779.
We have compared the steroid responsiveness of Müller glial cells of intact embryonic chicken retina with that of primary cultures derived from Müller glia. Appropriately constructed fusion genes were found to be highly glucocorticoid inducible after their cotransfection with an expression vector encoding the human glucocorticoid receptor (GR) into intact embryonic d-10 (E10) or E5.5 retina. Dramatically attenuated inductions were obtained after contransfection of Müller cell primary cultures. The progesterone receptor (PR) was also demonstrated to function in intact retina, but not in Müller cell primary cultures. An immunochemical assay was utilized to confirm that a glucocorticoid-responsive, beta-galactosidase-encoding fusion gene was specifically induced in Müller cells after its transfection into intact retina. Thus, in contrast to Müller cells in intact retina, Müller cells in primary culture have lost the capacity to achieve transcriptional activation by steroid receptors. We postulate that coordinate expression of the GR, and other more general factors required for steroid inducibility, is lost by dispersion and primary culture of retinal Müller glial cells.
我们比较了完整胚胎鸡视网膜中 Müller 胶质细胞与源自 Müller 胶质细胞的原代培养物对类固醇的反应性。在将适当构建的融合基因与编码人糖皮质激素受体(GR)的表达载体共转染到完整的胚胎第 10 天(E10)或 E5.5 视网膜后,发现这些融合基因具有高度的糖皮质激素诱导性。在 Müller 细胞原代培养物共转染后,诱导作用显著减弱。还证明了孕酮受体(PR)在完整视网膜中起作用,但在 Müller 细胞原代培养物中不起作用。利用免疫化学分析证实,将编码β-半乳糖苷酶的糖皮质激素反应性融合基因转染到完整视网膜后,在 Müller 细胞中特异性诱导表达。因此,与完整视网膜中的 Müller 细胞相比,原代培养中的 Müller 细胞已失去通过类固醇受体实现转录激活的能力。我们推测,视网膜 Müller 胶质细胞的分散和原代培养导致 GR 以及类固醇诱导所需的其他更普遍因子的协调表达丧失。