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豚鼠肾上腺中源自两种阿片肽前体的免疫反应性肽的共定位及特性研究

Co-localization and characterization of immunoreactive peptides derived from two opioid precursors in guinea pig adrenal glands.

作者信息

Evans C J, Erdelyi E, Hunter J, Barchas J D

出版信息

J Neurosci. 1985 Dec;5(12):3423-7. doi: 10.1523/JNEUROSCI.05-12-03423.1985.

Abstract

Peptides derived from both proenkephalin and prodynorphin have been identified in guinea pig adrenal medulla. In extracts of whole adrenal glands radioimmunoassays directed to the prodynorphin-derived peptides alpha-neoendorphin, dynorphin A, and dynorphin B detected high concentrations of immunoreactive material ranging from 113 to 216 pmol/gm. The concentrations measured by radioimmunoassays directed to the proenkephalin products met-enkephalin-Arg-Gly-Leu and met-enkephalin-Arg-Phe were 878 and 484 pmol/gm, respectively. No metorphamide or dynorphin(1-8) could be detected in the adrenals. Leucine-enkephalin immunoreactivity which can be generated from either prodynorphin or proenkephalin could also be measured in the extracts. Gel filtration showed the immunoreactive material, with the exception of that measured by the alpha-neoendorphin radioimmunoassay, to be predominantly of high molecular weight ranging from Mr = 3,000 to 12,000. Immunocytochemistry, using well characterized antisera to alpha-neoendorphin and met-enkephalin-Arg-Gly-Leu, demonstrated that the prodynorphin and proenkephalin products were present in the same cells in the medulla region of the gland. The results show that two opioid peptide precursors can be localized in the same cells and exhibit some common features in their processing. As a relatively homogeneous, localized system, the guinea pig adrenal gland should prove a valuable, in vivo model for the study of co-localized opioid precursors.

摘要

在豚鼠肾上腺髓质中已鉴定出源自前脑啡肽原和前强啡肽原的肽类。在整个肾上腺提取物中,针对源自前强啡肽原的肽类α-新内啡肽、强啡肽A和强啡肽B的放射免疫测定法检测到高浓度的免疫反应性物质,范围为113至216 pmol/g。针对前脑啡肽原产物甲硫氨酸脑啡肽-精氨酸-甘氨酸-亮氨酸和甲硫氨酸脑啡肽-精氨酸-苯丙氨酸的放射免疫测定法所测得的浓度分别为878和484 pmol/g。在肾上腺中未检测到甲硫氨酰胺或强啡肽(1-8)。提取物中也可测量到可由前强啡肽原或前脑啡肽原产生的亮氨酸脑啡肽免疫反应性。凝胶过滤显示,除α-新内啡肽放射免疫测定法所测量的物质外,免疫反应性物质主要为高分子量,范围从Mr = 3000至12000。使用针对α-新内啡肽和甲硫氨酸脑啡肽-精氨酸-甘氨酸-亮氨酸的特征明确的抗血清进行免疫细胞化学分析表明,前强啡肽原和前脑啡肽原产物存在于肾上腺髓质区域的同一细胞中。结果表明,两种阿片肽前体可定位于同一细胞中,并在其加工过程中表现出一些共同特征。作为一个相对同质的局部系统,豚鼠肾上腺应被证明是研究共定位阿片肽前体的有价值的体内模型。

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