Immunohistological Study of Monkey Foveal Retina
Immunohistological Study of Monkey Foveal Retina
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DOI：
10.1038/s41598-019-41793-y
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发表时间：
2019-03-27
影响因子：
4.6
通讯作者：
Takai, Shinji
中科院分区：
文献类型：
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作者：
Ikeda, Tsunehiko;Nakamura, Kimitoshi;Takai, Shinji
中央凹是位于黄斑中心的解剖学凹坑，无血管、缺氧，具有其他组织干细胞巢的特征。我们假设在中央凹中可能存在未分化的视网膜干细胞样细胞，并且可能发生神经发生。因此，我们使用食蟹猴视网膜进行了免疫组织学研究。制备包括中心凹的视网膜冷冻组织切片后，对胶质纤维酸性蛋白（GFAP）、巢蛋白、波形蛋白、神经元特异性 III 类 β-微管蛋白（Tuj-1）、视紫红质抑制蛋白 4、神经丝、CD117、CD44、Ki67 和细胞视黄醛结合蛋白（CRALBP）进行免疫染色，然后进行荧光和/或共聚焦显微镜检查。组织切片的免疫染色能够清楚地观察到对应于中央凹 Muller 细胞锥内半层的强 GFAP 阳性细胞。中心凹斜面的表层部分与 GFAP 和波形蛋白共染色。在中心凹视网膜最内层观察到 Tuj-1 阳性细胞，该细胞跨越周围的神经节细胞层。此外，在中央凹处观察到 Tuj-1 和 GFAP 的共定位。在中央凹的光感受器间基质中发现了 CD117 和 CD44 的共表达。小凹锥体的巢蛋白和视紫红质抑制蛋白 4 均呈阳性染色，然而，小凹外侧的光感受器层对巢蛋白的染色较弱。在亨利层的内半部观察到巢蛋白和波形蛋白的共定位，而在外半部主要观察到巢蛋白和神经丝的共定位。在小凹周围的外丛状层和神经节细胞层的细胞突起中观察到分散的Ki67阳性细胞。大部分 GFAP 阳性区域的 CRALBP 免疫染色呈阴性。 Muller细胞锥分为内层的GFAP强阳性细胞（可能是星形胶质细胞）和外层的巢蛋白阳性/GFAP弱阳性放射状胶质细胞样细胞。这些发现表明，小凹中的此类未分化细胞群可能参与维持形态和再生。
The fovea centralis, an anatomically concave pit located at the center of the macula, is avascular, hypoxic, and characteristic of stem-cell niches of other tissues. We hypothesized that in the fovea, undifferentiated retinal-stem-cell-like cells may exist, and that neurogenesis may occur. Hence, we performed an immunohistological study using cynomolgus monkey retinas. After preparing frozen tissue sections of the retina including the foveal pit, immunostaining was performed for glial fibrillary acidic protein (GFAP), nestin, vimentin, neuron-specific class III beta-tubulin (Tuj-1), arrestin 4, neurofilament, CD117, CD44, Ki67, and cellular retinaldehyde-binding protein (CRALBP), followed by fluorescence and/or confocal microscopy examinations. Immunostaining of the tissue sections enabled clear observation of strongly GFAP-positive cells that corresponded to the inner-half layer of the foveolar Muller cell cone. The surface layer of the foveal slope was partially costained with GFAP and vimentin. Tuj-1-positive cells were observed in the innermost layer of the foveolar retina, which spanned to the surrounding ganglion cell layer. Moreover, colocalization of Tuj-1 and GFAP was observed at the foveal pit. The coexpression of CD117 and CD44 was found in the interphotoreceptor matrix of the fovea. The foveolar cone stained positive for both nestin and arrestin 4, however, the photoreceptor layer outside of the foveola displayed weak staining for nestin. Colocalization of nestin and vimentin was observed in the inner half of the Henle layer, while colocalization of nestin and neurofilament was observed in the outer half, predominantly. Scattered Ki67-positive cells were observed in the cellular processes of the outer plexiform layer and the ganglion cell layer around the foveola. Immunostaining for CRALBP was negative in most parts of the GFAP-positive area. The Muller cell cone was divided into GFAP-strongly positive cells, presumably astrocytes, in the inner layer and nestin-positive/GFAP-weakly positive radial glia-like cells in the outer layer. These findings indicated that groups of such undifferentiated cells in the foveola might be involved in maintaining morphology and regeneration.