Calof A L, Reichardt L F
Neurosci Lett. 1985 Aug 30;59(2):183-9. doi: 10.1016/0304-3940(85)90197-1.
Purified motoneurons from chick, cultured on polycationic substrata treated with myotube-conditioned (MCM), respond by rapidly extending neurites. When MCM, partially purified by salt precipitation and ion-exchange chromatography, was fractionated on Sepharose CL-4B, the peak of neurite outgrowth-promoting activity (NOPA) corresponded to a peak of laminin (LA) immunoreactivity. Fractions from this peak contained a protein band that comigrated with an LA standard on sodium dodecyl sulfate gels. Antibodies to LA immunoprecipitated all motoneuron NOPA from MCM, and the specifically immunoprecipitated material comigrated with LA in both reducing and non-reducing gels. It thus appears that LA in MCM is essential for the ability of this conditioned medium to promote motoneuron neurite outgrowth.
从鸡中纯化的运动神经元,培养在经肌管条件培养基(MCM)处理的聚阳离子基质上时,会通过快速延伸神经突做出反应。当通过盐沉淀和离子交换色谱法部分纯化的MCM在琼脂糖凝胶CL-4B上进行分级分离时,神经突生长促进活性(NOPA)的峰值与层粘连蛋白(LA)免疫反应性的峰值相对应。该峰值的组分含有一条在十二烷基硫酸钠凝胶上与LA标准品迁移率相同的蛋白带。针对LA的抗体从MCM中免疫沉淀了所有运动神经元NOPA,并且在还原和非还原凝胶中,特异性免疫沉淀的物质都与LA迁移率相同。因此,MCM中的LA对于这种条件培养基促进运动神经元神经突生长的能力至关重要。