Newsome D A, Rothman R J
Lions Eye Research Laboratories, Louisiana State University Medical Center School of Medicine, New Orleans 70112.
Invest Ophthalmol Vis Sci. 1987 Nov;28(11):1795-9.
Zinc, an essential trace element, is present in unusually high concentrations in the chorioretinal complex relative to most other tissues. Because little has been known about the interactions between the retinal pigment epithelium and free or protein-associated zinc, we studied 65Zn uptake by human retinal pigment epithelium in vitro. When monolayers were exposed to differing concentrations from 0 to 30 microM 65Zn in Dulbecco's modified Eagle's medium with 5.4 gm/l glucose at 37 degrees C and 4 degrees C, we observed a temperature-dependent saturable accumulation of the radiolabel. With 15 microM 65Zn, we saw a biphasic pattern of uptake with a rapid first phase and a slower second phase over 120 min. Uptake of 65Zn was inhibited by iodacetate and cold, and reduced approximately 50% by the addition of 2% albumin to the labelling medium. Neither ouabain nor 2-deoxyglucose inhibited uptake. Cells previously exposed to 65Zn retained approximately 70% of accumulated 65Zn 60 min after being changed to radiolabel-free medium. Following removal of cells from the extracellular matrix adherent to the dish bottom, a variable amount of nonspecific binding of 65Zn to the residual matrix was demonstrated. These observations are consistent with a facilitated type of transport and demonstrate the ability of human retinal pigment epithelium in vitro to accumulate and retain zinc.
锌作为一种必需的微量元素,相对于大多数其他组织,在脉络膜视网膜复合体中的浓度异常高。由于人们对视网膜色素上皮与游离或与蛋白质结合的锌之间的相互作用了解甚少,我们在体外研究了人视网膜色素上皮对65锌的摄取。当单层细胞在含有5.4克/升葡萄糖的杜尔贝科改良伊格尔培养基中,于37℃和4℃下暴露于0至30微摩尔/升的不同浓度65锌时,我们观察到放射性标记物呈温度依赖性饱和积累。在15微摩尔/升65锌的情况下,我们在120分钟内观察到摄取呈双相模式,第一阶段快速,第二阶段较慢。65锌的摄取受到碘乙酸盐和低温的抑制,在标记培养基中添加2%白蛋白可使摄取减少约50%。哇巴因和2-脱氧葡萄糖均不抑制摄取。先前暴露于65锌的细胞在更换为无放射性标记的培养基60分钟后,仍保留约70%积累的65锌。从附着在培养皿底部的细胞外基质中去除细胞后,证明65锌与残留基质存在可变数量的非特异性结合。这些观察结果与一种易化转运类型一致,并证明人视网膜色素上皮在体外具有积累和保留锌的能力。