POSTNATAL PRECESSING OF PRO DYNORPAHIN IN THE BRAIN

强啡肽原是三种阿片肽的共同前体：强啡肽 强啡肽B和α-新内啡肽。 虽然有相当多的 关于这些肽的解剖分布的信息， 的前体，以及由此衍生的终产物的序列， 前体，相对较少的是知道产后个体发育这一点 系统，翻译后处理中涉及的步骤， 前强啡肽，或调节区域特异性蛋白水解的机制 强啡肽原的加工过程 本建议将考虑两个问题：1） 大细胞/后部强啡肽原的出生后个体发生 垂体系统和黑质纹状体系统; 2）确定是否 从下丘脑和纹状体获得的出生后神经元是合适的 用于体外单层组织培养研究。 最初的研究将监测稳态水平的 垂体后叶中的强啡肽原相关终产物， 黑质在出生后的发展。 这些研究将涉及 放射免疫分析结合凝胶过滤色谱法 和高效液相色谱法。 体外研究将测试 不同年龄新生神经元在体外单层培养中的存活率 脉冲放射性标记范例。 将通过免疫亲和法进行分析 SDS聚丙烯酰胺凝胶柱层析 电泳

Pro-dynorphin is the common precursor for three opioid peptides: dynorphin A, dynorphin B, and alpha-neo-endorphin. Although there is considerable information on the anatomical distribution of these peptides, the structure of the precursor, and the sequences of the end products derived from this precursor, relatively little is known about the postnatal ontogeny of this system, the steps involved in the post-translational processing of pro-dynorphin, or the mechanisms for regulating region specific proteolytic processing of pro-dynorphin. This proposal will consider two issues: 1) the postnatal ontogeny of pro-dynorphin in the magnocellular/posterior pituitary system and the striato-nigral system; 2) determine whether postnatal neurons obtained from the hypothalamus and striatum are suitable for in vitro monolayer tissue culture studies. The initial studies will monitor the steady state levels of pro-dynorphin-related end products in the posterior pituitary and substantia nigra during postnatal development. These studies will involve radioimmunoassay analyses in conjunction with gel filtration chromatography and high performance liquid chromatography. The in vitro studies will test the viability of different aged neonatal neurons in monolayer culture with a pulse radiolabeling paradigm. Analyses will be done by immunoaffinity column chromatography in conjunction with SDS polyacrylamide gel electrophoresis.