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新鲜分离的人及猴视网膜色素上皮细胞中的二氢吡啶敏感性钙电流。

Dihydropyridine-sensitive calcium currents in freshly isolated human and monkey retinal pigment epithelial cells.

作者信息

Ueda Y, Steinberg R H

机构信息

Department of Ophthalmology, University of California, San Francisco School of Medicine 94143-0730.

出版信息

Invest Ophthalmol Vis Sci. 1995 Feb;36(2):373-80.

PMID:7843908
Abstract

PURPOSE

The authors previously reported that rat retinal pigment epithelial (RPE) cells exhibit an ionic current through voltage-operated calcium channels that is dihydropyridine sensitive. They attempted to record the same current from freshly isolated fetal or adult primate RPE cells, as well as from cultured cells.

METHODS

The whole-cell version of the patch-clamp technique was applied to RPE cells freshly isolated by enzymatic dissociation from fetal human and adult human and monkey eyes, as well as cultured fetal and adult human RPE cells. The cells were loaded with cesium to minimize potassium-related current and were bathed in an extracellular solution containing 40 mM barium to intensify the calcium currents.

RESULTS

Freshly isolated cells, both fetal and adult, showed sustained and inward-going barium current through voltage-operated calcium channels with membrane depolarizations from a high holding voltage of -60 mV. This current was affected by dihydropyridine compounds. Cultured human RPE cells showed no sign of a calcium current of this type.

CONCLUSIONS

Freshly isolated fetal and adult human RPE cells, as well as adult monkey, exhibit calcium current through voltage-operated, dihydropyridine-sensitive channels, similar to the neuronal L-type, just as in rat RPE cells.

摘要

目的

作者先前报道,大鼠视网膜色素上皮(RPE)细胞通过对二氢吡啶敏感的电压门控钙通道表现出离子电流。他们试图从新鲜分离的胎儿或成年灵长类动物RPE细胞以及培养的细胞中记录相同的电流。

方法

采用膜片钳技术的全细胞模式,对通过酶解从胎儿人类、成年人类和猴子眼睛新鲜分离的RPE细胞以及培养的胎儿和成年人类RPE细胞进行研究。细胞内充灌铯以最小化与钾相关的电流,并置于含有40 mM钡的细胞外溶液中以增强钙电流。

结果

新鲜分离的胎儿和成年细胞,在从 -60 mV的高钳制电压进行膜去极化时,通过电压门控钙通道显示出持续的内向钡电流。该电流受二氢吡啶化合物影响。培养的人类RPE细胞未显示出这种类型钙电流的迹象。

结论

新鲜分离的胎儿和成年人类RPE细胞以及成年猴子的RPE细胞,通过电压门控、对二氢吡啶敏感的通道表现出钙电流,类似于神经元L型,与大鼠RPE细胞一样。

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