Kanan Yogita, Wicker Lea D, Al-Ubaidi Muayyad R, Mandal Nawajes A, Kasus-Jacobi Anne
Department of Cell Biology, Universit of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.
Invest Ophthalmol Vis Sci. 2008 Mar;49(3):1071-8. doi: 10.1167/iovs.07-1207.
RDH11 and RDH12 are closely related retinol dehydrogenases expressed in the retina. RDH12 has been linked to the early-onset retinal dystrophy Leber congenital amaurosis, whereas RDH11 has not been associated with human disease. To understand their physiological roles, the authors investigated their expression during development and their regulation by light-induced oxidative stress in mouse retina.
Quantitative RT-PCR and immunoblot analysis were used for quantification of RDH11 and RDH12 during development and oxidative stress. Expression during development was measured between embryonic day (E) 12 and postnatal day (P) 210 (7 months) in C57BL/6 mouse eyes. Expression during light-induced oxidative stress was measured between 2 and 24 hours of exposure to light in BALB/c mouse retina.
The RDH11 level was low and remarkably constant during development and oxidative stress. RDH12 expression started at P7 and increased until P30 to approximately sevenfold higher than RDH11. Oxidative stress induced by exposure to constant bright light led to a rapid and significant decrease of RDH12 protein.
The low and constant expression of RDH11 suggested a housekeeping function for this enzyme. The onset of RDH12 expression during the maturation of photoreceptor cells suggested a function related to the visual process. The light-induced rapid decrease of RDH12 protein, preceding the decrease of the mRNA, suggested a specific degradation of the protein rather than a regulation of gene expression.
RDH11和RDH12是在视网膜中表达的密切相关的视黄醇脱氢酶。RDH12与早发性视网膜营养不良——莱伯先天性黑蒙相关,而RDH11尚未与人类疾病相关联。为了解它们的生理作用,作者研究了它们在发育过程中的表达以及在小鼠视网膜中由光诱导的氧化应激对其的调节。
采用定量逆转录聚合酶链反应(qRT-PCR)和免疫印迹分析来定量RDH11和RDH12在发育和氧化应激过程中的表达。在C57BL/6小鼠眼中,测量胚胎期第12天(E12)至出生后第210天(P210,7个月)发育过程中的表达。在BALB/c小鼠视网膜中,测量光照2至24小时期间光诱导氧化应激过程中的表达。
RDH11水平在发育和氧化应激过程中较低且相当稳定。RDH12的表达在P7开始,并持续增加直至P30,比RDH11高约7倍。持续强光照射诱导的氧化应激导致RDH12蛋白迅速且显著减少。
RDH11的低水平且稳定的表达表明该酶具有管家功能。感光细胞成熟过程中RDH12表达的开始表明其功能与视觉过程相关。光诱导的RDH12蛋白在mRNA减少之前迅速减少,表明是蛋白质的特异性降解而非基因表达的调节。