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缝隙连接蛋白连接子 43(GJA1)在人视网膜和视神经中的免疫定位。

Immunolocalization of gap junction protein connexin43 (GJA1) in the human retina and optic nerve.

机构信息

Department of Ophthalmology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.

出版信息

Invest Ophthalmol Vis Sci. 2010 Aug;51(8):4028-34. doi: 10.1167/iovs.09-4847. Epub 2010 Apr 7.

Abstract

PURPOSE

Gap junctions are intercellular channels that have been implicated in the pathogenesis of neuronal death after central nervous system injury. This study determines the expression pattern of gap junction protein connexin43 in the human retina and optic nerve.

METHODS

An affinity-isolated polyclonal antibody to the C-terminal segment of the cytoplasmic domain of human connexin43 was used to determine connexin43 localization. Postmortem human eyes were examined by immunohistochemical staining of frozen sections using antibodies to connexin43. Antibody binding was detected using confocal microscopy and fluorochrome-conjugated secondary antibodies. Double-label immunohistochemistry identified the cell types expressing connexin43.

RESULTS

Connexin43 immunoreactivity was detected in the human retina on glial fibrillary acidic protein (GFAP)-positive astrocytes in the retinal ganglion cell layer and, to a lesser extent, on the processes of glutamine synthetase-labeled Müller cells. The retinal and choroidal circulations showed strong connexin43 immunolabeling. Dense connexin43 immunoreactivity was present between adjacent cells of the retinal pigment epithelium, and there was diffuse connexin43 immunoreactivity on GFAP-positive astrocytes in the optic nerve.

CONCLUSIONS

In the human retina and optic nerve, connexin43 is present on glia, blood vessels, and epithelial cells. An understanding of the distribution of connexin43 in the normal retina and optic nerve may be used to evaluate changes associated with retinal and optic nerve disease.

摘要

目的

缝隙连接是细胞间的通道,已被牵涉到中枢神经系统损伤后神经元死亡的发病机制中。本研究旨在确定缝隙连接蛋白 connexin43 在人视网膜和视神经中的表达模式。

方法

使用针对人 connexin43 胞质域 C 末端片段的亲和分离的多克隆抗体来确定 connexin43 的定位。通过对死后人眼的冰冻切片进行免疫组织化学染色,使用 connexin43 抗体来检测 connexin43 的结合。使用共聚焦显微镜和荧光素标记的二级抗体检测抗体结合。双标免疫组化鉴定表达 connexin43 的细胞类型。

结果

在人视网膜中,connexin43 免疫反应性可在神经节细胞层的胶质纤维酸性蛋白 (GFAP) 阳性星形胶质细胞上以及程度较轻的谷氨酰胺合成酶标记的 Müller 细胞的突起上检测到。视网膜和脉络膜循环显示出强烈的 connexin43 免疫标记。视网膜色素上皮细胞之间存在密集的 connexin43 免疫反应性,视神经中 GFAP 阳性星形胶质细胞上存在弥漫性 connexin43 免疫反应性。

结论

在人视网膜和视神经中,connexin43 存在于神经胶质细胞、血管和上皮细胞上。了解 connexin43 在正常视网膜和视神经中的分布情况,可能有助于评估与视网膜和视神经疾病相关的变化。

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