Variation and immunocyochemical localization of processing enzymes in proopiomelanocortin cells during ontoneny

原阿黑皮素细胞在色调过程中加工酶的变化和免疫细胞化学定位

基本信息

批准号：
04670002
负责人：
TANAKA Shigeyasu
金额：
$ 1.28万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670002/
关键词：
Processing Processing enzyme Secretory granule Pituitary gland Immunocyochemistry 内分泌細胞

项目摘要

PC1/PC3 and PC2 are endoproteases involved in prohormone cleavage at pairs of basic amino acids. To determine the cellular and subcellular distribution of PC1/PC3 and PC2 in the pituitary glands, we generated their polyclonal antisera in rabbits, using two synthetic peptide antigens corresponding to amino acids 442-459 (ST-28) of PC1/PC3 and 613-629 (ST-29) of PC2 and two bacterially expressed antigens covering amino acids 145-414 (KN-1) of PC1/PC3 and 385-637 (KN-2) of PC2. Western blot analysis revealed the presence of PC1/PC3 87 and 68 kDa) and PC2 (70 kDa) in the rat pancreatic islets, indicating that each of the antisera was specific for the corresponding antigen. In the present immunocyochemical studies, many neuroendcrine cells displayd positive reaction to these antibodies. In almost all the animals as far as examined, PC1/PC3 and PC2 showed differential immunoreactivity in proopiomelanocortin (POMC) cells. In the anterior lobe, antiserum against PC1/PC3 exclusively immunostained corticotrophs in the intermediate lobe. Immunoelectron microscopy, using double-labeling with gold particles of different sizes revealed that almost all of immunolabeled ACTH- or alpha-MSH-secretory granules were also positive to PC1/PC3 and PC2 antibody. This differential localization of PC1/PC3 and PC2 provides a novel argument for the subtrate-specificity of the PC1/PC3 and PC2 enzymes in the processing of pituitary POMC cells. To elucidate factors that regulate changes in the processing patterns of POMC in the pituitary corticotrophs during postnatal development, in near further, the presence of PC1/PC3 and PC2 will be examined immunocytochemically.

PC1/PC3和PC2是在基本氨基酸对的激素裂解中涉及的内螺旋。为了确定垂体中PC1/PC3和PC2的细胞和亚细胞分布，我们使用两个合成肽抗原在兔子中生成了它们的多克隆抗血清，与PC1/PC3和613-629（ST-29）的PC1/PC3酸氨基酸442-459（ST-28）相对应的氨基酸442-459（ST-28）和PC2和613-629（ST-29）（ST-29）（ST-29）（ST-29）（ST-29）（ST-29） PC1/PC3的145-414（KN-1）和PC2的385-637（KN-2）。 Western印迹分析显示，大鼠胰岛中存在PC1/PC3 87和68 kDa）和PC2（70 kDa），表明每个抗血清都针对相应的抗原具有特异性。在目前的免疫化学研究中，许多神经内脑细胞对这些抗体表现出阳性反应。在所检查的几乎所有动物中，PC1/PC3和PC2在亚蛋白酶素（POMC）细胞中均显示出差异性免疫反应性。在前叶中，针对PC1/PC3的抗血清在中间叶中仅免疫染色的皮质营养素。免疫电子显微镜，使用具有不同大小的金颗粒的双标记表明，几乎所有免疫标记的ACTH或α-MSH-MSH分泌颗粒也对PC1/PC3和PC2抗体呈阳性。 PC1/PC3和PC2的这种差异定位为垂体POMC细胞的处理中PC1/PC3和PC2酶的亚酸特异性提供了新的论点。为了阐明在产后发育过程中垂体皮质营养物中POMC加工模式变化的因素，在接近进一步的情况下，将在PC1/PC3和PC2的存在下进行免疫细胞化学检查。