Das S R, Bhardwaj N, Gouras P
Department of Ophthalmology, Columbia University, New York, NY 10032.
Biochem J. 1990 May 15;268(1):201-6. doi: 10.1042/bj2680201.
The synthesis and release of 11-cis-retinoids by primary cultures of human retinal pigment epithelium (RPE) and the transfer of these retinoids to co-incubated human rod outer segments (ROS) were studied. Monolayers of 2-3-week-old cultured RPE incorporate tritiated all-trans-retinol, esterify it to the corresponding retinyl palmitate, form 11-cis-retinol and 11-cis-retinaldehyde and release retinaldehyde into the culture medium. The ratio of 11-cis to all-trans isomers of retinol, retinyl palmitate and retinaldehyde formed in the cells along with retinaldehyde released and incorporated into the ROS progressively increases, indicating a progressive increase in the concentration of 11-cis isomer from the time it is formed in RPE cells until its transfer to ROS. Incorporation of 11-cis-retinaldehyde into the ROS is directly related to the amount of albumin present in the media, suggesting the transfer of retinoids from RPE to photoreceptor to be a protein-mediated process. Events leading to isomerization, esterification, oxidation and release of retinoids by human RPE and incorporation of retinoids into ROS can therefore be examined in vitro.
研究了人视网膜色素上皮(RPE)原代培养物中11-顺式视黄醛的合成与释放,以及这些视黄醛向共孵育的人视杆外段(ROS)的转移。2 - 3周龄培养的RPE单层细胞摄取氚标记的全反式视黄醇,将其酯化为相应的棕榈酸视黄酯,形成11-顺式视黄醇和11-顺式视黄醛,并将视黄醛释放到培养基中。细胞中形成的视黄醇、棕榈酸视黄酯和视黄醛的11-顺式与全反式异构体的比例,以及释放并掺入ROS中的视黄醛,都逐渐增加,这表明从11-顺式异构体在RPE细胞中形成到其转移至ROS的过程中,11-顺式异构体的浓度逐渐增加。11-顺式视黄醛掺入ROS与培养基中白蛋白的含量直接相关,这表明视黄醛从RPE向光感受器的转移是一个蛋白质介导的过程。因此,可以在体外研究导致人RPE对视黄醛进行异构化、酯化、氧化和释放以及视黄醛掺入ROS的过程。