Eiden L E, Siegel R E, Giraud P, Brenneman D E
Unit on Molecular and Cellular Neurobiology, Laboratory of Cell Biology, NIMH, Bethesda, MD 20892.
Brain Res Dev Brain Res. 1988 Nov 1;44(1):141-50. doi: 10.1016/0165-3806(88)90125-3.
The ontogeny of vasoactive intestinal polypeptide (VIP), and Met-enkephalin in primary cultures of spinal cord/dorsal root ganglia from 12-day mouse embryos was examined by radioimmunoassay and immunohistochemistry. Met-enkephalin levels rose from less than 5 to 700 pg/culture over 26 days and were half maximal by day 16-18 in culture. VIP levels rose from less than 1 to 30 pg/culture over the same period, but were already half maximal by day 9. Met-enkephalin immunoreactivity was localized in multipolar medium sized neurons while VIP immunoreactivity was visualized both in neurons with extensively branched processes and in bipolar cells some of which appeared to be dorsal root ganglion cells. Tetrodotoxin (TTX)-sensitive spontaneous release of both peptides developed in parallel with the ability to stimulate peptide release with elevated potassium. Factors affecting the ontogeny of neuropeptide expression in, and release from, spinal cord neurons can now be examined in vitro in a strictly defined neurochemical environment.
通过放射免疫测定法和免疫组织化学方法，对来自12日龄小鼠胚胎的脊髓/背根神经节原代培养物中血管活性肠多肽（VIP）和甲硫氨酸脑啡肽的个体发生进行了研究。在26天的时间里，甲硫氨酸脑啡肽的水平从每培养物少于5 pg上升到700 pg，在培养的第16 - 18天达到最大值的一半。同期，VIP水平从每培养物少于1 pg上升到30 pg，但在第9天就已达到最大值的一半。甲硫氨酸脑啡肽免疫反应性定位于多极中等大小的神经元中，而VIP免疫反应性在具有广泛分支突起的神经元以及双极细胞中都可见，其中一些双极细胞似乎是背根神经节细胞。两种肽对河豚毒素（TTX）敏感的自发释放与用高钾刺激肽释放的能力平行发展。现在可以在严格定义的神经化学环境中，在体外研究影响脊髓神经元中神经肽表达个体发生和释放的因素。
