DiMaggio D A, Chronwall B M, Buchanan K, O'Donohue T L
Neuroscience. 1985 Aug;15(4):1149-57. doi: 10.1016/0306-4522(85)90259-3.
Radioimmunoassay was combined with high pressure liquid chromatography and immunohistochemistry to establish the identity of pancreatic polypeptide-like immunoreactive material in the central nervous system of the rat. Antisera to avian pancreatic polypeptide, bovine pancreatic polypeptide, the invariant amidated carboxyterminal hexapeptide fragment of mammalian pancreatic polypeptides and the structurally related peptide, neuropeptide Y, were used immunocytochemically to localize neurons containing immunoreactive pancreatic polypeptide-like material in rat brain. Adjacent brain sections stained by the indirect immunofluorescent technique and single sections from double-staining experiments demonstrated that identical fibers and perikarya stained for pancreatic polypeptide-like immunoreactive material by antisera directed against each of the four peptides. Characterization of pancreatic polypeptide-like immunoreactive material in chromatographed rat brain extracts by radioimmunoassay using antisera to either neuropeptide Y or the carboxy-terminal portion of the pancreatic polypeptide molecule revealed that the major peak of immunoreactive material, as measured by either assay, appeared to co-elute with synthetic porcine neuropeptide Y. A minor peak of immunoreactive material co-eluting with peptide YY standard was indicated by the neuropeptide Y radioimmunoassay. This was contrasted by data obtained from chromatographic profiles of rat pancreas, which showed that the main immunoreactive peak, as measured by the neuropeptide Y assay, co-eluted with porcine peptide YY, with a minor peak co-eluting with porcine neuropeptide Y. The main peak of immunoreactive material in pancreas, as measured by the pancreatic polypeptide carboxy-terminal radioimmunoassay, eluted considerably earlier than standard peptide YY, neuropeptide Y and bovine pancreatic polypeptide, and is probably identical to rat pancreatic polypeptide.(ABSTRACT TRUNCATED AT 250 WORDS)
放射免疫测定法与高压液相色谱法及免疫组织化学相结合,以确定大鼠中枢神经系统中胰多肽样免疫反应性物质的特性。使用针对禽胰多肽、牛胰多肽、哺乳动物胰多肽不变的酰胺化羧基末端六肽片段以及结构相关肽神经肽Y的抗血清,通过免疫细胞化学方法定位大鼠脑中含有胰多肽样免疫反应性物质的神经元。通过间接免疫荧光技术染色的相邻脑切片以及双重染色实验的单一切片表明,针对这四种肽中的每一种的抗血清对胰多肽样免疫反应性物质染色的纤维和核周体是相同的。通过使用针对神经肽Y或胰多肽分子羧基末端部分的抗血清进行放射免疫测定,对经色谱分离的大鼠脑提取物中的胰多肽样免疫反应性物质进行表征,结果显示,两种测定方法所测得的免疫反应性物质的主要峰似乎与合成猪神经肽Y共洗脱。神经肽Y放射免疫测定法表明存在一个与肽YY标准品共洗脱的免疫反应性物质小峰。这与从大鼠胰腺色谱图获得的数据形成对比,后者显示,神经肽Y测定法所测得的主要免疫反应性峰与猪肽YY共洗脱,一个小峰与猪神经肽Y共洗脱。通过胰多肽羧基末端放射免疫测定法测得的胰腺中免疫反应性物质的主要峰,比标准肽YY、神经肽Y和牛胰多肽洗脱得早得多,可能与大鼠胰多肽相同。(摘要截短至250字)