Bridges C D
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907.
Vision Res. 1989;29(12):1711-7. doi: 10.1016/0042-6989(89)90153-3.
Ocular tissue homogenates were incubated in darkness with [11,12-3H] all-trans retinol. Formation of radiolabeled 11-cis retinol was used as an index of isomerase activity and was determined by high-performance liquid chromatography. Isomerase was found in the eyes of cattle, human, rat, chicken, turtle, goldfish and frog, representing the mammals, birds, reptiles, bony fishes and amphibians. The enzyme was concentrated in the pigment epithelium (RPE). Variable activity was found in the retina, where the amount of radiolabeled 11-cis retinol formed under standard incubation conditions at protein concentrations of 0.03-1.08 mg/ml was 6.4 +/- 6.0% of that in the RPE-choroid. Using the same methodology, we could not detect isomerase in the retinas of three cephalopods (Octopus, Sepia and Loligo). In rats, isomerase was present at postnatal day 10 but not at postnatal days 0 and 4. Therefore, the expression in the RPE of retinol isomerase, which is essential for the formation of rhodopsin in the developing photoreceptors, is coordinated with the emergence of the rod outer segment in the retina. However, the continued expression of this enzyme in RCS rats does not depend on the presence of photoreceptors, because loss of photoreceptors was not associated with an absence of isomerase activity in RCS rats. Our findings suggest that a reciprocal flow of retinoids between the retina and the site of isomerase action in the RPE is a feature common to the visual cycle in all vertebrates.
将眼组织匀浆与[11,12 - 3H]全反式视黄醇在黑暗中孵育。放射性标记的11 - 顺式视黄醇的形成用作异构酶活性的指标,并通过高效液相色谱法测定。在牛、人、大鼠、鸡、龟、金鱼和青蛙的眼中发现了异构酶,分别代表哺乳动物、鸟类、爬行动物、硬骨鱼和两栖动物。该酶集中在色素上皮(RPE)中。在视网膜中发现了可变活性,在标准孵育条件下,蛋白质浓度为0.03 - 1.08 mg/ml时,视网膜中形成的放射性标记的11 - 顺式视黄醇的量是RPE - 脉络膜中的6.4±6.0%。使用相同的方法,我们在三种头足类动物(章鱼、乌贼和枪乌贼)的视网膜中未检测到异构酶。在大鼠中,出生后第10天存在异构酶,但出生后第0天和第4天不存在。因此,视黄醇异构酶在RPE中的表达与视网膜中视杆外段的出现相协调,而视黄醇异构酶对于发育中的光感受器中视紫红质的形成至关重要。然而,该酶在RCS大鼠中的持续表达并不依赖于光感受器的存在,因为在RCS大鼠中光感受器的丧失与异构酶活性的缺失无关。我们的研究结果表明,视网膜与RPE中异构酶作用部位之间类视黄醇的双向流动是所有脊椎动物视觉循环的一个共同特征。