Functional characterization of mouse RDH11 as a retinol dehydrogenase involved in dark adaptation in vivo
Functional characterization of mouse RDH11 as a retinol dehydrogenase involved in dark adaptation in vivo
复制标题
DOI：
10.1074/jbc.m413789200
复制
发表时间：
2005-05-27
影响因子：
4.8
通讯作者：
Edwards, AO
中科院分区：
文献类型：
--
作者：
Kasus-Jacobi, A;Ou, JF;Edwards, AO
我们先前克隆了小鼠RDH 11（mRDH 11）作为由转录因子固醇调节元件结合蛋白调节的基因，并表明它是在非眼组织如肝脏和睾丸以及视网膜中表达的视黄醇脱氢酶（Kasus-Jacobi，A.，欧，杰，Bashmakov，Y. K.，谢尔顿，J.M.，理查德森，J. A.，戈尔茨坦，J.L.，Brown，M. S.（2003）J.Biol.Chem.278，32380-32389）。有人提出，它在漂白光光异构化后的视觉发色团11-顺式-视黄醛的再循环中起作用，这一途径被称为视觉循环。在这项工作中，我们描述了我们对mRDH 11的眼功能的研究。我们通过用lacZ报告基因替换mrdh 11编码序列来创建基因敲除小鼠用于表达谱分析。5-溴-4-氯-3-吲哚基-β-D-吡喃半乳糖苷（X-Gal）染色显示该基因在感光细胞中活跃转录。我们通过免疫印迹分析表明，mRDH 11与从视网膜的非外段部分纯化的视网膜膜相关。RDH 11缺陷小鼠在发育和衰老过程中未发现明显的视网膜缺损。通过视网膜电图研究mRDH 11破坏的功能后果。与野生型小鼠相比，暗适应延迟了2.5-3倍。然而，11-顺式-视网膜循环动力学在暗适应过程中没有受到影响，这表明mRDH 11不参与视觉周期。我们认为，mRDH 11中断影响视色素代谢的感光细胞内节和延迟的动力学暗适应通过调节钙稳态。
We previously cloned mouse RDH11 ( mRDH11) as a gene regulated by the transcription factor sterol regulatory element-binding proteins and showed that it is a retinol dehydrogenase expressed in non-ocular tissues such as the liver and testis and in the retina (Kasus-Jacobi, A., Ou, J., Bashmakov, Y. K., Shelton, J. M., Richardson, J. A., Goldstein, J. L., and Brown, M. S. ( 2003) J. Biol. Chem. 278, 32380-32389). It was proposed to function in the recycling of the visual chromophore 11-cis-retinal after photoisomerization by a bleaching light, a pathway referred to as the visual cycle. In this work, we describe our studies on the ocular function of mRDH11. We created a knockout mouse by replacing the mrdh11 coding sequence with the lacZ reporter gene for expression profiling. 5-Bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining demonstrated active transcription of this gene in photoreceptor cells. We show by immunoblot analysis that mRDH11 is associated with retinal membranes purified from a non-outer segment fraction of the retina. No obvious retinal defect was found during development and aging of RDH11-deficient mice. The functional consequences of mRDH11 disruption were investigated by electroretinography. Dark adaptation was delayed by a factor of 2.5-3 compared with wild-type mice. However, the kinetics of 11-cis-retinal recycling during dark adaptation was not affected, suggesting that mRDH11 is not involved in the visual cycle. We propose that mRDH11 disruption affects retinoid metabolism in photoreceptor inner segments and delays the kinetics of dark adaptation through modulation of calcium homeostasis.