Kanan Yogita, Kasus-Jacobi Anne, Moiseyev Gennadiy, Sawyer Kjell, Ma Jian-Xing, Al-Ubaidi Muayyad R
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Exp Eye Res. 2008 Feb;86(2):344-54. doi: 10.1016/j.exer.2007.11.006. Epub 2007 Nov 17.
To determine whether cones and Müller cells in the rod dominated retina cooperate to regenerate the 11-cis retinal chromophore via the retinoid cycle, two cell lines from the rod dominated retinas of Murine were used for this study: 661W, a mouse cell line derived from cones, and rMC-1, a rat Müller cell line. Retinoid cycle enzymes were analyzed by RT-PCR, and their catalytic activity was detected by incubation with retinoids and analyzed by HPLC. We found that 661W cells are capable of reducing all-trans retinal to all-trans retinol due to the presence of multiple dehydrogenases and to generate minor amounts of retinyl-ester. The rMC-1 cells take up all-trans retinol and oxidize it to all-trans retinal or esterify it to retinyl-ester, but are incapable of isomerizing all-trans retinoids to 11-cis retinoids. This could be a reflection of lack of necessary activities in Müller cells in vivo, which suggests that Müller cells do not contribute to retinoid cycling by regenerating 11-cis retinoids. Alternatively, this could be due to the potential that rMC-1, as a transformed cell line, has stopped expressing the proteins needed for the regeneration of 11-cis retinoids.
为了确定在视杆细胞主导的视网膜中,视锥细胞和穆勒细胞是否通过类视黄醇循环协同再生11-顺式视黄醛发色团,本研究使用了来自小鼠视杆细胞主导的视网膜的两种细胞系:661W,一种源自视锥细胞的小鼠细胞系,以及rMC-1,一种大鼠穆勒细胞系。通过RT-PCR分析类视黄醇循环酶,并通过与类视黄醇孵育检测其催化活性,然后用HPLC进行分析。我们发现,由于存在多种脱氢酶,661W细胞能够将全反式视黄醛还原为全反式视黄醇,并生成少量视黄酯。rMC-1细胞摄取全反式视黄醇并将其氧化为全反式视黄醛或将其酯化生成视黄酯,但无法将全反式类视黄醇异构化为11-顺式类视黄醇。这可能反映了体内穆勒细胞缺乏必要的活性,这表明穆勒细胞不会通过再生11-顺式类视黄醇来参与类视黄醇循环。或者,这可能是由于rMC-1作为一种转化细胞系,可能已经停止表达再生11-顺式类视黄醇所需的蛋白质。