Effects of astroglia on the development of cultured neurons from embryonic rat cerebral cortex.

Y Ibata, S Toya, S Kohsaka
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Abstract

1. Previous studies from our laboratory demonstrated that subcultured astroglia enhance neurite outgrowth and survival of cultured neurons from embryonic rat cerebral cortex, but suppress proliferation of neuroblasts. 2. In the present study, the mechanisms of these three effects were further investigated. 3. Dissociated neurons were seeded on poly-L-lysine-coated coverslips which were plated on subcultured astroglia, and the survival, proliferation and neurite outgrowth of the neurons were investigated. Under these conditions, survival and antimitotic effects were also observed, while neurite extension was not stimulated. 4. The results clearly indicate that neuronal survival and proliferation are regulated by soluble factors produced by astroglia. 5. We also postulated that the neurite-promoting effect of astroglia is mediated by cell-cell contact. 6. This idea was confirmed by the finding that neurite extension was enhanced when the neurons were cultured directly on heat-treated astroglia. 7. The neurite-promoting effect was found to be specific to astroglia. 8. We preliminarily characterized the astroglial surface neurite-promoting factors (ASNPFs). 9. The relationship of laminin to ASNPFs was examined by using antibody to laminin. Laminin antibody did not inhibit the ASNPF activity. 10. The effect of digestion of heat-treated astroglia with enzymes (sialidase and endo-beta-galactosidase) on the ASNPF activity was also examined. 11. These enzyme treatments did not inhibit the ASNPF activity. 12. These results suggest that enhancement of the neurite-promoting activity is not associated with the sugar moiety of ASNPFs.

星形胶质细胞对胚胎大鼠大脑皮层培养神经元发育的影响。
1. 我们实验室之前的研究表明,继代星形胶质细胞能促进胚胎大鼠大脑皮层神经元的神经突生长和存活,但抑制神经母细胞的增殖。2. 本研究进一步探讨了这三种效应的作用机制。3.将解离的神经元接种于聚赖氨酸包被盖片上,盖片镀于传代星形胶质细胞上,观察神经元的存活、增殖和神经突生长情况。在这些条件下,还观察到存活和抗有丝分裂作用,而神经突的延伸不受刺激。4. 结果表明,星形胶质细胞产生的可溶性因子可调节神经元的存活和增殖。5. 我们还假设星形胶质细胞的神经突促进作用是通过细胞间接触介导的。6. 当神经元直接在热处理的星形胶质细胞上培养时,神经突的延伸得到了增强,这一发现证实了这一观点。7. 神经突促进作用被发现是星形胶质细胞特有的。8. 我们初步鉴定了星形胶质表面神经突促进因子(ASNPFs)。9. 用层粘连蛋白抗体检测层粘连蛋白与asnpf的关系。层粘连蛋白抗体对ASNPF活性无抑制作用。10. 研究了唾液酸酶和内切-半乳糖苷酶对热处理星形胶质细胞ASNPF活性的影响。11. 这些酶处理没有抑制ASNPF的活性。12. 这些结果表明,促进神经突活性的增强与asnpf的糖部分无关。
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