Smith R G, Vaca K, McManaman J, Appel S H
J Neurosci. 1986 Feb;6(2):439-47. doi: 10.1523/JNEUROSCI.06-02-00439.1986.
In vivo labeling of motoneurons with retrogradely transported wheat germ agglutinin, and in situ staining of cholinergic neurons with a monoclonal antibody against CAT, have been used as methods to identify motoneurons in cultures of dissociated 14 d embryonic rat ventral spinal cord. With these two different labeling techniques, we demonstrate that skeletal muscle extracts maintain motoneuron survival, and induce motoneuron morphologic and cholinergic differentiation in vitro. Further, we show that several partially purified muscle extract-derived proteins separately augment each of these specific parameters of motoneuron development. A 55,000 Da neutral glycoprotein specifically induces both ACh synthesis and motoneuron-selective process outgrowth and elongation, while a 1200-1500 Da peptide selectively augments both apparent motoneuron survival and ACh synthesis. A third trophic agent, a 33,000-35,000 Da acidic glycoprotein, induces process outgrowth in both motoneurons and nonmotoneurons of the ventral spinal cord, but has no effect on ACh synthesis.
用逆行运输的小麦胚凝集素对运动神经元进行体内标记，并用抗胆碱乙酰转移酶单克隆抗体对胆碱能神经元进行原位染色，已被用作鉴定14日龄胚胎大鼠腹侧脊髓解离培养物中运动神经元的方法。通过这两种不同的标记技术，我们证明骨骼肌提取物能维持运动神经元的存活，并在体外诱导运动神经元的形态和胆碱能分化。此外，我们表明几种部分纯化的肌肉提取物衍生蛋白分别增强了运动神经元发育的这些特定参数中的每一个。一种55000道尔顿的中性糖蛋白特异性地诱导乙酰胆碱合成以及运动神经元选择性的突起生长和延伸，而一种1200 - 1500道尔顿的肽选择性地增强明显的运动神经元存活和乙酰胆碱合成。第三种营养因子，一种33000 - 35000道尔顿的酸性糖蛋白，诱导腹侧脊髓运动神经元和非运动神经元的突起生长，但对乙酰胆碱合成没有影响。
