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花生凝集素结合鸡视网膜中的糖蛋白:它们存在于米勒胶质细胞中。

Peanut agglutinin binding glycoproteins in the chick retina: their presence in Müller glia cells.

作者信息

Arregui C, Barra H S, Landa C A

机构信息

Centro de Investigaciones en Química Biológica de Córdoba, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Argentina.

出版信息

J Neurosci Res. 1992 Mar;31(3):532-42. doi: 10.1002/jnr.490310317.

Abstract

The histological and cellular distribution and some biochemical characteristics of components that bind peanut agglutinin (PNA), a lectin that recognizes preferentially terminal galactose-beta (1-3) N-acetyl galactosamine disaccharide residues of glycoconjugates, were studied in chick retinal tissue and in dissociated retinal cells after their differentiation in culture. In sections of retinal tissue from animals 7 days after hatching (Rp7), in addition to the inner and outer segments of the photoreceptor layer, the plexiform and optic fiber layers were stained with rhodamine-labeled PNA, indicating that, besides photoreceptor cells, other cellular types contribute to the PNA staining. We present evidence indicating that at least part of this staining is provided by Müller glia cells. In cultures of dissociated cells from retinas at embryonic day 7 (R7), photoreceptor-like cells and flat Müller glia-derived cells but not neurons were stained with rhodamine-labeled PNA. Furthermore, Müller glia cells isolated from Rp7 were also brightly stained with PNA. Western blot assays of extracts from R7 showed the presence of PNA binding glycoproteins of 31-33 kDa and a component that migrates at the dye front. In addition to the components detected in R7, extracts from R14 and Rp7 showed the presence of a major PNA binding glycoprotein of 175 kDa and a minor glycoprotein of 220 kDa. Extracts from the photoreceptor layer contain the 175 and 220 kDa glycoproteins, indicating their association with photoreceptor cells. The 31-33 kDa components were detected in extracts from the remnant inner retina, suggesting their association with the Müller glia cells. Supporting this view, these components and not those of 175 and 220 kDa were detected in cell cultures enriched in flat Müller glia-derived cells. Only the 31-33 kDa components and the component that migrates at the dye front were detected in extracts from cell cultures enriched in photoreceptor-like cells, suggesting the need of some environmental element for the expression of the 175 and 220 kDa components in the differentiated photoreceptor cells.

摘要

花生凝集素(PNA)是一种凝集素,优先识别糖缀合物的末端半乳糖-β(1-3)N-乙酰半乳糖胺二糖残基。本文研究了PNA结合成分在雏鸡视网膜组织以及培养分化后的解离视网膜细胞中的组织学和细胞分布,以及一些生化特性。在孵化后7天(Rp7)的动物视网膜组织切片中,除了光感受器层的内段和外段,神经纤维层和视神经纤维层也被罗丹明标记的PNA染色,这表明除了光感受器细胞外,其他细胞类型也对PNA染色有贡献。我们提供的证据表明,这种染色至少部分是由米勒胶质细胞提供的。在胚胎第7天(R7)视网膜解离细胞的培养物中,光感受器样细胞和平坦的米勒胶质细胞衍生细胞被罗丹明标记的PNA染色,但神经元未被染色。此外,从Rp7分离的米勒胶质细胞也被PNA强烈染色。R7提取物的蛋白质免疫印迹分析显示存在31 - 33 kDa的PNA结合糖蛋白和一种迁移到染料前沿的成分。除了在R7中检测到的成分外,R14和Rp7的提取物还显示存在一种主要的175 kDa的PNA结合糖蛋白和一种次要的220 kDa糖蛋白。光感受器层的提取物含有175和220 kDa的糖蛋白,表明它们与光感受器细胞相关。在残留的视网膜内层提取物中检测到31 - 33 kDa的成分,表明它们与米勒胶质细胞相关。支持这一观点的是,在富含平坦的米勒胶质细胞衍生细胞的细胞培养物中检测到了这些成分,而未检测到175和220 kDa的成分。在富含光感受器样细胞的细胞培养物提取物中,仅检测到31 - 33 kDa的成分和迁移到染料前沿的成分,这表明在分化的光感受器细胞中表达175和220 kDa的成分需要一些环境因素。

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