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人视网膜母细胞瘤细胞系WERI-Rb1中神经元相关III类β-微管蛋白亚型(hβ4)和微管相关蛋白2(MAP2)的抗原表达。一项比较免疫印迹和免疫细胞化学研究。

Antigenic expression of neuron-associated class III beta-tubulin isotype (h beta 4) and microtubule-associated protein 2 (MAP2) by the human retinoblastoma cell line WERI-Rb1. A comparative immunoblot and immunocytochemical study.

作者信息

Gass P, Frankfurter A, Katsetos C D, Herman M M, Donoso L A, Rubinstein L J

机构信息

Department of Pathology, University of Virginia School of Medicine, Charlottesville.

出版信息

Ophthalmic Res. 1990;22(1):57-66. doi: 10.1159/000267002.

Abstract

The antigenic expression of two neuron-associated microtubule proteins, class III beta-tubulin isotype (h beta 4) and microtubule-associated protein 2 (MAP2), was evaluated in a comparative immunoblot and immunocytochemical study of the human retinoblastoma cell line WERI-Rb1 maintained for up to 30 days in three different in vitro conditions. Western blots were performed on whole sodium dodecyl sulfate extracts of cells grown in floating suspensions, on Gelfoam matrices and on coverslips. Immunoperoxidase histochemistry was performed on matrix cultures. Immunoblotting demonstrated that h beta 4 and MAP2 were present under all culture conditions. By immunocytochemistry, staining of cytologically undifferentiated cells with anti-h beta 4 and anti-MAP2 monoclonal antibodies was found on Gelfoam matrix explants. In contrast, glial fibrillary acidic protein was not detected by either immunoblots or immunocytochemistry. These findings are in keeping with the solely neuroblastic nature of this line and provide no evidence for its divergent (i.e. neuronal and glial) differentiation capacity.

摘要

在一项比较免疫印迹和免疫细胞化学研究中,对人视网膜母细胞瘤细胞系WERI-Rb1在三种不同体外条件下培养长达30天的情况进行了评估,以检测两种神经元相关微管蛋白,即III类β-微管蛋白亚型(hβ4)和微管相关蛋白2(MAP2)的抗原表达。对在悬浮培养、明胶海绵基质和盖玻片上生长的细胞的全十二烷基硫酸钠提取物进行了蛋白质免疫印迹分析。对基质培养物进行了免疫过氧化物酶组织化学分析。免疫印迹显示,在所有培养条件下均存在hβ4和MAP2。通过免疫细胞化学方法,在明胶海绵基质外植体上发现,用抗hβ4和抗MAP2单克隆抗体对未分化细胞进行染色。相比之下,无论是免疫印迹还是免疫细胞化学都未检测到胶质纤维酸性蛋白。这些发现与该细胞系仅具有成神经细胞性质相符,并未提供其具有不同分化能力(即神经元和胶质细胞分化能力)的证据。

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