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利用共聚焦显微镜免疫组织化学技术对人视网膜的细微结构进行定义。

Fine structure of the human retina defined by confocal microscopic immunohistochemistry.

机构信息

Eye & ENT Hospital, College of Medicine, Fudan University , Shanghai, China.

Shanghai Key Laboratory of Visual Impairment and Restoration, Fudan University , Shanghai, China.

出版信息

Br J Biomed Sci. 2021 Jan;78(1):28-34. doi: 10.1080/09674845.2020.1776586. Epub 2020 Jul 17.

DOI:10.1080/09674845.2020.1776586
PMID:32498649
Abstract

INTRODUCTION

Research in to the pathophysiology of the complex layers of retinal and sub-retinal cells is hampered by inadequate recognition of particular cells and tissues. A comprehensive panel of antibodies recognising retinal tissues is lacking. Our purpose was to determine the value of a panel of antibodies labelling various cells in the human retina.

METHOD

Five groups of antibodies labelled frozen sections of retinas: (1) protein kinase C-α, Glutamine Synthetase (GS) and ionized calcium-binding adapter molecule 1 (Iba1); (2) Parvalbumin, Calretinin and glial fibrillary acidic protein (GFAP); (3) Thy1, GS and Iba1; (4) Rhodopsin, GS and Iba1; and (5) Brn3a, Rhodopsin and protein kinase C-α. The distribution of these antigens were determined by confocal microscopy and calculated grey value of each antibody in each layer of the retina by Image J.

RESULTS

Different antibodies determined certain retinal layers. Thy 1 is a good determinant of the ganglion cell layer, whilst GS is present in all layers except the photoreceptor layer. Brn3a is specific for the ganglion cell layer whilst parvalbumin marks the ganglion cell layer and the outer plexiform layer. Rhodopsin strongly marks the photoreceptor layer, but this is also marked weakly by GFAP.

CONCLUSION

The multiple labelling of human retinal cells brings further understanding of the biological characteristics and functions of these cells, and provides a theoretical basis for their possible role in diseases. In the growing field of human retina research, our data may provide a point of reference for future studies of the human retina.

摘要

简介

对视网膜和视网膜下细胞复杂层次的病理生理学的研究受到对特定细胞和组织认识不足的阻碍。缺乏能识别视网膜组织的全面抗体组。我们的目的是确定标记人视网膜中各种细胞的抗体组的价值。

方法

五组抗体标记了冷冻的视网膜切片:(1)蛋白激酶 C-α、谷氨酰胺合成酶 (GS) 和离子钙结合衔接分子 1 (Iba1);(2)副甲状腺蛋白、钙结合蛋白和神经胶质纤维酸性蛋白 (GFAP);(3)Thy1、GS 和 Iba1;(4)视紫红质、GS 和 Iba1;和(5)Brn3a、视紫红质和蛋白激酶 C-α。通过共聚焦显微镜确定这些抗原的分布,并通过 Image J 计算视网膜各层中每种抗体的灰度值。

结果

不同的抗体确定了某些视网膜层。Thy1 是神经节细胞层的良好标志物,而 GS 存在于除感光细胞层之外的所有层中。Brn3a 特异性标记神经节细胞层,而副甲状腺蛋白标记神经节细胞层和外丛状层。视紫红质强烈标记感光细胞层,但 GFAP 也能弱标记。

结论

对人视网膜细胞的多重标记进一步了解了这些细胞的生物学特征和功能,并为它们在疾病中的可能作用提供了理论基础。在人类视网膜研究的不断发展领域,我们的数据可能为未来的人类视网膜研究提供参考点。

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