Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
Int J Mol Sci. 2022 Jan 26;23(3):1408. doi: 10.3390/ijms23031408.
One important role of epigenetic regulation is controlling gene expression in development and homeostasis. However, little is known about epigenetics' role in regulating opsin expression. Cell cultures (HEK 293, Y79, and WERI) producing different levels of opsins were treated with 5-aza-2'-deoxycytidine (5-Aza-dc) and/or sodium butyrate (SB) or suberoylanilide hydroxamic acid (SAHA) for 72 h. Global DNA methylation, site-specific methylation, and expressions of opsins were measured by LUMA assay, bisulfite pyrosequencing, and qPCR, respectively. Mouse retinal explants from wild-type P0/P1 pups were ex vivo cultured with/without 5-Aza-dc or SAHA for 6 days. The morphology of explants, DNA methylation, and expressions of opsins was examined. The drugs induced global DNA hypomethylation or increased histone acetylation in cells, including DNA hypomethylation of rhodopsin () and L-opsin () and a concomitant increase in their expression. Further upregulation of and/or in HEK 293 or WERI cells was observed with 5-Aza-dc and either SB or SAHA combination treatment. Mouse retinal explants developed normally but had drug-dependent differential DNA methylation and expression patterns of opsins. DNA methylation and histone acetylation directly regulate opsin expression both in vitro and ex vivo. The ability to manipulate opsin expression using epigenetic modifiers enables further study into the role of epigenetics in eye development and disease.
表观遗传调控的一个重要作用是控制发育和内稳态中的基因表达。然而，对于表观遗传在调节视蛋白表达中的作用知之甚少。用 5-氮杂-2′-脱氧胞苷（5-Aza-dc）和/或丁酸钠（SB）或琥珀酰亚胺基羟肟酸（SAHA）处理产生不同水平视蛋白的细胞培养物（HEK 293、Y79 和 WERI）72 小时。通过 LUMA 测定、亚硫酸氢盐焦磷酸测序和 qPCR 分别测量全基因组 DNA 甲基化、特定位点甲基化和视蛋白的表达。将野生型 P0/P1 幼鼠的视网膜外植体在有/无 5-Aza-dc 或 SAHA 的情况下进行离体培养 6 天。检查外植体的形态、DNA 甲基化和视蛋白的表达。这些药物诱导细胞中的全基因组 DNA 低甲基化或组蛋白乙酰化增加，包括视紫红质（）和 L-视蛋白（）的 DNA 低甲基化，以及它们表达的相应增加。在用 5-Aza-dc 联合 SB 或 SAHA 处理后，在 HEK 293 或 WERI 细胞中观察到 和/或 的进一步上调。发育正常的小鼠视网膜外植体，但具有药物依赖性的视蛋白 DNA 甲基化和表达模式差异。DNA 甲基化和组蛋白乙酰化直接调节视蛋白的体外和离体表达。使用表观遗传修饰剂来操纵视蛋白表达的能力使我们能够进一步研究表观遗传在眼睛发育和疾病中的作用。
