Takayanagi M, Watanabe T
Department of Veterinary Anatomy, School of Agriculture, Nagoya University, Japan.
Tissue Cell. 1996 Feb;28(1):17-24. doi: 10.1016/s0040-8166(96)80040-1.
The colocalization of aromatic L-amino acid decarboxylase (AADC), dopamine beta-hydroxylase (DBH), S-100 protein and chromogranin A (CgA) in the insulin-containing B-cells of the chicken endocrine pancreas was investigated by using light microscopic immunohistochemistry and combined pre-embedding immunoperoxidase and post-embedding immunogold electron microscopic immunocytochemistry. Using the post-embedding method, immunoreactivity against the anti-insulin serum by protein A-gold technique was observed in the core of all types of B-cell granules. Immunoreaction with anti-S-100 protein serum was detected in the core of all types of B-cell granules. Immunoreaction with anti-S-100 protein serum was detected in the core of all types of B-cell granules from non-osmicated tissues even by post-embedding method, but immunoreactivities against the anti-AADC, DBH and CgA sera were only demonstrable in crystalloid granules of B-cells by pre-embedding method. Immunoreaction with the anti-CgA serum was also detected in the cytoplasmic matrix around crystalloid granules and also in the dense bodies showing immunonegative with anti-insulin serum. From these results, it seems likely that S-100 protein co-stored all types of B-cell granules involved in the maturation of granules, and AADC, DBH and CgA are related to the synthesis of noradrenaline in crystalloid granules of B-cells.
运用光学显微镜免疫组织化学以及预包埋免疫过氧化物酶和后包埋免疫金电子显微镜免疫细胞化学技术,对鸡内分泌胰腺含胰岛素的B细胞中芳香族L-氨基酸脱羧酶(AADC)、多巴胺β-羟化酶(DBH)、S-100蛋白和嗜铬粒蛋白A(CgA)的共定位进行了研究。采用后包埋法,通过蛋白A-金技术在所有类型B细胞颗粒的核心部位观察到了抗胰岛素血清的免疫反应性。在所有类型B细胞颗粒的核心部位均检测到了与抗S-100蛋白血清的免疫反应。即使采用后包埋法,在未经锇酸处理的组织中所有类型B细胞颗粒的核心部位也检测到了与抗S-100蛋白血清的免疫反应,但通过预包埋法,仅在B细胞的晶体颗粒中证实了与抗AADC、DBH和CgA血清的免疫反应性。在晶体颗粒周围的细胞质基质以及对抗胰岛素血清呈免疫阴性的致密小体中也检测到了与抗CgA血清的免疫反应。从这些结果来看,S-100蛋白似乎共同储存于参与颗粒成熟的所有类型B细胞颗粒中,而AADC、DBH和CgA与B细胞晶体颗粒中去甲肾上腺素的合成有关。