Bloch-Gallego E, Huchet M, el M'Hamdi H, Xie F K, Tanaka H, Henderson C E
UPR 8402 du CNRS-U 249 de l'INSERM, Montpellier, France.
Development. 1991 Jan;111(1):221-32. doi: 10.1242/dev.111.1.221.
Motoneurons were identified in vitro by a new method using the SC1 monoclonal antibody. They constituted up to 30% of total neurons in cultures of whole spinal cord from 4.5-day chicken embryos, and survived for at least 5 days in the presence of muscle extract, but not in its absence. By contrast, other neurons and floor-plate cells survived without muscle-derived factors. Motoneurons were purified to homogeneity by 'panning' on dishes coated with SC1 antibody; they developed rapidly even in the absence of other spinal cells. Concentrations of muscle extract required for half-maximal motoneuron survival were indistinguishable in pure and mixed cultures, suggesting that muscle-derived factors act directly on motoneurons. Other purified growth factors tested, including ciliary neurotrophic factor, did not have the survival-promoting activity of muscle.
利用SC1单克隆抗体的新方法在体外鉴定出运动神经元。在来自4.5天大鸡胚的全脊髓培养物中,它们占总神经元的比例高达30%,并且在有肌肉提取物存在的情况下能存活至少5天,而在没有肌肉提取物时则不能存活。相比之下,其他神经元和底板细胞在没有肌肉衍生因子的情况下也能存活。通过在包被有SC1抗体的培养皿上进行“淘选”,运动神经元被纯化至同质;即使在没有其他脊髓细胞的情况下,它们也能快速发育。在纯培养物和混合培养物中,运动神经元半数最大存活所需的肌肉提取物浓度没有差异,这表明肌肉衍生因子直接作用于运动神经元。所测试的其他纯化生长因子,包括睫状神经营养因子,均不具有肌肉的促存活活性。