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昆虫麻蝇(Leucophaea maderae)中一种胰岛素相关肽的免疫细胞化学定位及免疫化学特性分析

Immunocytochemical localization and immunochemical characterization of an insulin-related peptide in the insect Leucophaea maderae.

作者信息

Hansen G N, Hansen B L, Jørgensen P N, Scharrer B

机构信息

Institute of Cell Biology and Anatomy, University of Copenhagen, Denmark.

出版信息

Cell Tissue Res. 1990 Feb;259(2):265-73. doi: 10.1007/BF00318448.

Abstract

Immunocytochemical tests with eight monoclonal antibodies against either bovine or human insulin and seven polyclonal antibodies against bovine insulin were carried out to determine the presence of insulin-like neuropeptides in the brain and affiliated neuroendocrine structures of the insect Leucophaea maderae. Reaction products identified in the brain, subesophageal ganglion, and corpus cardiacum-corpus allatum complex indicate the presence of materials resembling mammalian insulins in its antigenic properties. The immunostaining observed with monoclonal antibodies appears to indicate the occurrence of an insulin-related peptide that shows sequential similarities with parts of both the A- and B-chains of mammalian insulin molecules. These suppositions are supported by the results of dot-blot and two-site time-resolved immunofluorometric assay (TRI-IFMA) screenings of fractions of Leucophaea tissue extracts obtained by chromatography. The polyclonal antibodies yielded reaction products in some of the same areas and in additional parts of the neuroendocrine system not visualized by the monoclonal antibodies. Immunoreaction was observed in the following areas: the pars intercerebralis of the protocerebrum, the nervi corporis cardiaci I transporting insulin-like material to the corpus cardiacum, the dorsolateral protocerebral area and the optic lobes, the deutocerebrum, the tritocerebrum, and the subesophageal ganglion. In addition, smaller cell bodies with immunoreactive deposits occur at the border between proto- and deuto-cerebrum, and in the central area of the protocerebrum. The distribution of reactive material in the corpus cardiacum-corpus allatum complex after use of both groups of antibodies was the same.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

利用8种抗牛胰岛素或人胰岛素的单克隆抗体以及7种抗牛胰岛素的多克隆抗体进行免疫细胞化学试验,以确定昆虫麻蝇大脑及附属神经内分泌结构中是否存在胰岛素样神经肽。在大脑、咽下神经节以及心侧体 - 咽侧体复合体中鉴定出的反应产物表明,存在抗原特性与哺乳动物胰岛素相似的物质。用单克隆抗体观察到的免疫染色似乎表明存在一种与胰岛素相关的肽,它与哺乳动物胰岛素分子的A链和B链部分具有序列相似性。通过色谱法获得的麻蝇组织提取物各组分的斑点印迹和双位点时间分辨免疫荧光测定(TRI - IFMA)筛选结果支持了这些推测。多克隆抗体在一些相同区域以及单克隆抗体未显示的神经内分泌系统其他部分产生了反应产物。在以下区域观察到免疫反应:前脑的脑间部、将胰岛素样物质输送到心侧体的第一心神经、背外侧前脑区域和视叶、中脑、后脑以及咽下神经节。此外,在原脑和中脑之间的边界以及原脑的中央区域出现了有免疫反应性沉积物的较小细胞体。使用两组抗体后,心侧体 - 咽侧体复合体中反应物质的分布相同。(摘要截短于250字)

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