Marek K L, Mains R E
Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
J Neurochem. 1989 Jun;52(6):1807-16. doi: 10.1111/j.1471-4159.1989.tb07261.x.
The biosynthesis of neuropeptide Y (NPY) and norepinephrine (NE) has been examined in dissociated neuronal cultures from newborn rat superior cervical ganglion (SCG). NPY synthetic rate was measured by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis after incubation in medium containing a labeled amino acid. The authenticity of the NPY was confirmed by reverse-phase HPLC analyses of tryptic peptides. The NPY synthetic rate in cultures grown in complete serum free medium increased 30-fold after plating, in parallel to catecholamine synthesis; both NPY and the catecholamines reached the rate for adult SCG neurons. This development in culture is seen without spinal cord input, target organs, or significant numbers of glial cells. NPY synthesis was maintained in the face of a major decrease in the rate of NE production after cholinergic induction.
已对新生大鼠颈上神经节(SCG)解离的神经元培养物中神经肽Y(NPY)和去甲肾上腺素(NE)的生物合成进行了研究。在含有标记氨基酸的培养基中孵育后,通过免疫沉淀和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测量NPY合成速率。通过对胰蛋白酶肽段进行反相高效液相色谱分析,证实了NPY的真实性。在完全无血清培养基中生长的培养物中,接种后NPY合成速率增加了30倍,与儿茶酚胺合成平行;NPY和儿茶酚胺均达到成年SCG神经元的合成速率。在没有脊髓输入、靶器官或大量神经胶质细胞的情况下,培养物中出现了这种发育情况。在胆碱能诱导后NE产生速率大幅下降的情况下,NPY合成仍得以维持。