Hatten M E, Furie M B, Rifkin D B
J Neurosci. 1982 Sep;2(9):1195-206. doi: 10.1523/JNEUROSCI.02-09-01195.1982.
The role of the matrix glycoprotein fibronectin in the formation of the external granular layer of the developing mouse cerebellum was investigated by in vitro studies of the binding of cerebellar cells to a fibronectin-coated culture substratum and by in vivo immunocytochemical localization of antiplasma fibronectin antiserum in cerebellar tissue. The adhesion of cells dissociated from embryonic and early postnatal mouse cerebellum is developmental stage-specific when the cells are plated on tissue culture substrata derivatized with human plasma fibronectin. Cells dissociated from mouse cerebellum at embryonic day 13 form cellular aggregates on insoluble plasma fibronectin. In contrast, cells dissociated from embryonic day 16 through postnatal day 7 cerebellum form a monolayer. Time-lapse video recordings reveal extensive cell movement of late embryonic and early postnatal cerebellar cells on insoluble plasma fibronectin. Late embryonic and early postnatal cerebellar cells bind to fibronectin but do not degrade the fibronectin substratum. Immunocytochemical studies of the binding of antiplasma fibronectin antisera to cryostat sections of intact embryonic and early postnatal cerebellar tissue reveal a brightly stained region of endogenous fibronectin along the route of granule cell migration from the lateral caudal part of the neuroepithelium lining the fourth ventricle up onto the external surface of the cerebellar anlage. When the formation of the external granular layer is completed, the intense region of fibronectin is no longer visible.
通过体外研究小脑细胞与纤连蛋白包被的培养基质的结合以及体内免疫细胞化学定位抗血浆纤连蛋白抗血清在小脑组织中的分布,来研究基质糖蛋白纤连蛋白在发育中小鼠小脑外颗粒层形成中的作用。当将从小鼠胚胎期和出生后早期小脑解离的细胞接种在用人血浆纤连蛋白衍生化的组织培养基质上时,细胞的黏附具有发育阶段特异性。在胚胎第13天从小鼠小脑解离的细胞在不溶性血浆纤连蛋白上形成细胞聚集体。相比之下,从胚胎第16天到出生后第7天的小脑解离的细胞形成单层。延时视频记录显示晚期胚胎和出生后早期小脑细胞在不溶性血浆纤连蛋白上有广泛的细胞运动。晚期胚胎和出生后早期小脑细胞与纤连蛋白结合,但不降解纤连蛋白基质。抗血浆纤连蛋白抗血清与完整胚胎和出生后早期小脑组织冰冻切片结合的免疫细胞化学研究显示,沿着颗粒细胞从第四脑室内衬神经上皮的尾侧外侧部分迁移到小脑原基外表面的路径,有一个内源性纤连蛋白染色明亮的区域。当外颗粒层形成完成时,纤连蛋白的强烈区域不再可见。