Isolation and culture of motoneurons from embryonic chicken spinal cords.
Isolation and culture of motoneurons from embryonic chicken spinal cords.
复制标题
鸡胚脊髓运动神经元的分离和培养。
DOI：
10.1073/pnas.76.7.3537
复制
发表时间：
1979
影响因子：
11.1
通讯作者：
Y. Shimada
中科院分区：
文献类型：
--
作者：
S. Masuko;H. Kuromi;Y. Shimada
描述了一种从17-18期（Hamburger和汉密尔顿编号）鸡胚脊髓中分离胆碱能α运动神经元的方法，此时已表明运动神经元退出有丝分裂周期，但其它类型的神经元和神经胶质仍处于有丝分裂状态。在10 μ M 1-β-D-阿拉伯呋喃糖基胞嘧啶存在下，将脊髓腹侧一半的片段孵育24小时，以消除分裂细胞，随后解离成单细胞悬浮液。已经获得的以下证据表明，这些细胞是神经元，似乎是α运动神经元：（i）它们对阿拉伯呋喃糖基胞嘧啶的致死作用有抗性，因此在17-18期是有丝分裂后的;（ii）当在体外生长时，它们表现出与腹侧脊髓神经元相似的形态学特征，包括能够被银、尼氏染色，pH 6.5-7.0的亚甲蓝活体染色和胆碱乙酰转移酶组织化学;（iii）它们具有高胆碱乙酰转移酶活性;（iv）它们能够与肌肉形成功能性突触。
A method is described for isolating cholinergic alpha motoneurons from the spinal cord of chicken embryos at stage 17-18 (Hamburger and Hamilton numbering), at the time when it has been shown that motoneurons withdraw from the mitotic cycle but neurons of other types and glia are still mitotic. Fragments of the ventral half of the spinal cord are incubated for 24 hr in the presence of 10 microM 1-beta-D-arabinofuranosylcytosine in order to eliminate dividing cells and are subsequently dissociated into a suspension of single cells. The following evidence has been obtained and suggests that these cells are neuronal and appear to be alpha motoneurons: (i) they are resistant to the lethal effect of arabinofuranosylcytosine, and thus are postmitotic at stage 17-18; (ii) when grown in vitro, they exhibit morphological characteristics similar to those of ventral spinal neurons, which include the ability to be stained with silver, Nissl, methylene blue vital stain at pH 6.5-7.0, and choline acetyltransferase histochemistry; (iii) they have high choline acetyltransferase activity; (iv) they are capable of forming functional synapses with muscle.