Mettling C, Gouin A, Robinson M, el M'Hamdi H, Camu W, Bloch-Gallego E, Buisson B, Tanaka H, Davies A M, Henderson C E
INSERM U.382, Biochimie CNRS-INSERM, Montpellier, France.
J Neurosci. 1995 Apr;15(4):3128-37. doi: 10.1523/JNEUROSCI.15-04-03128.1995.
We compared the survival requirements of early- and late-born motoneurons from E5 chicken spinal cord. Density gradient centrifugation followed by immunopanning using SC1 antibody allowed us to purify two size classes of motoneuron. Large motoneurons retained by 6.8% metrizamide were shown by BrdU labeling in ovo to be born on average 1.5 d earlier than the small motoneurons recovered from the metrizamide pellet. Large motoneurons were both biochemically and functionally more mature: they expressed higher levels of choline acetyltransferase and low-affinity neurotrophin receptor, and had an acute requirement for trophic support from muscle-derived factors. After 24 hr in culture in basal medium, all early-born motoneurons died, whereas 60% of late-born motoneurons survived. Small motoneurons can develop into large motoneurons in ovo, suggesting that they represent a general transitional stage in motoneuron development. Our results suggest that a defined period elapses between birth of a motoneuron and its acquisition of trophic dependence, possibly corresponding to the time required for target innervation. This property may have important consequences for the timing and regulation of developmental motoneuron death.
我们比较了E5期鸡脊髓中早出生和晚出生运动神经元的存活需求。通过密度梯度离心,随后使用SC1抗体进行免疫淘选,我们得以纯化出两种大小不同的运动神经元。通过卵内BrdU标记显示,被6.8%的甲泛葡胺保留的大型运动神经元平均比从甲泛葡胺沉淀中回收的小型运动神经元早1.5天出生。大型运动神经元在生化和功能上都更成熟:它们表达更高水平的胆碱乙酰转移酶和低亲和力神经营养因子受体,并且对肌肉衍生因子的营养支持有急性需求。在基础培养基中培养24小时后,所有早出生的运动神经元都死亡了,而60%的晚出生运动神经元存活了下来。小型运动神经元在卵内可发育成大型运动神经元,这表明它们代表了运动神经元发育中的一个一般过渡阶段。我们的结果表明,运动神经元出生与其获得营养依赖性之间存在一个确定的时间段,这可能与靶标神经支配所需的时间相对应。这一特性可能对发育性运动神经元死亡的时间和调节具有重要影响。