Grierson J P, Petroski R E, Ling D S, Geller H M
Department of Pharmacology University of Medicine & Dentistry of New Jersey (UMDNJ)-Robert Wood Johnson Medical School, Piscataway 08854.
Brain Res Dev Brain Res. 1990 Aug 1;55(1):11-9. doi: 10.1016/0165-3806(90)90100-d.
We have observed a heterogeneity in the ability of a monolayer of cultured rat astrocytes to support the attachment and growth of dissociated embryonic hypothalamic neurons in culture. Areas of the monolayer which have an uneven surface ('rocky' astrocytes) are poor substrates for neuronal attachment and neuritic outgrowth, while surrounding areas of the glial monolayer ('flat' astrocytes) support extensive neuronal growth. Astrocytes obtained from both neonatal cerebral cortex or hypothalamus displayed 'rocky' morphology. We utilized immunocytochemical techniques with antibodies directed against putative adhesion molecules to investigate the source of this heterogeneity. Antibodies against tenascin/cytotacin, fibronectin, laminin, N-CAM, thrombospondin, heparan sulfate proteoglycan, and the p185 protein product of the neu oncogene were employed in indirect-immunofluorescence experiments. We found that the difference in the surface properties of astrocytes appears to be correlated with the expression of the extracellular matrix molecule tenascin/cytotacin, but not with any of the other molecules we tested. Our data suggest that tenascin/cytotactin is inhibitory to neuronal attachment and process outgrowth in the developing nervous system.
我们观察到,培养的大鼠星形胶质细胞单层支持解离的胚胎下丘脑神经元在培养物中附着和生长的能力存在异质性。单层中表面不平的区域(“粗糙”的星形胶质细胞)是神经元附着和神经突生长的不良底物,而胶质单层的周围区域(“扁平”的星形胶质细胞)则支持广泛的神经元生长。从新生大鼠大脑皮层或下丘脑获得的星形胶质细胞呈现出“粗糙”的形态。我们利用针对假定粘附分子的抗体的免疫细胞化学技术来研究这种异质性的来源。在间接免疫荧光实验中使用了针对腱生蛋白/细胞腱蛋白、纤连蛋白、层粘连蛋白、神经细胞粘附分子(N-CAM)、血小板反应蛋白、硫酸乙酰肝素蛋白聚糖以及neu癌基因的p185蛋白产物的抗体。我们发现,星形胶质细胞表面特性的差异似乎与细胞外基质分子腱生蛋白/细胞腱蛋白的表达相关,而与我们测试的任何其他分子无关。我们的数据表明,腱生蛋白/细胞腱蛋白在发育中的神经系统中对神经元附着和突起生长具有抑制作用。