Wagner G F, Copp D H, Friesen H G
Department of Physiology, University of Manitoba, Winnipeg, Canada.
Endocrinology. 1988 May;122(5):2064-70. doi: 10.1210/endo-122-5-2064.
This report describes the characterization of an antiserum to salmon teleocalcin, a glycoprotein hormone that has recently been isolated from sockeye salmon, Oncorhynchus nerka, corpuscles of Stannius (CS). Immunodiffusion studies showed that teleocalcin was immunologically identical in four species of Pacific salmon. Histological staining of adjacent sections of sockeye salmon CS by periodic acid-Schiff reagent and immunocytochemistry revealed that the teleocalcin-immunoreactive cells were also periodic acid-Schiff positive. In bioassays using rainbow trout (Salmo gairdneri), the biological effects of teleocalcin were completely abolished by pretreatment with the antiserum before injection. The antiserum was ineffective by itself, however, in neutralizing endogenous teleocalcin. Immunoprecipitation studies of in vitro translation products from salmon CS poly(A)+ RNA identified a 26K protein, which corresponds to the size of the teleocalcin monomer as predicted by cDNA sequencing studies. Similar banding patterns were obtained with purified teleocalcin and crude CS extracts by Western blot analysis. However, there was one extra band (32K) in the CS extracts that was not present in the purified hormone preparations. This may represent the pro-form of the hormone in salmon.
本报告描述了一种针对鲑鱼远钙素的抗血清的特性,远钙素是一种糖蛋白激素,最近从红大麻哈鱼(Oncorhynchus nerka)的斯坦尼氏小体(CS)中分离得到。免疫扩散研究表明,远钙素在四种太平洋鲑鱼中具有免疫同一性。用高碘酸 - 希夫试剂对红大麻哈鱼CS的相邻切片进行组织学染色和免疫细胞化学分析显示,远钙素免疫反应性细胞也是高碘酸 - 希夫阳性。在使用虹鳟(Salmo gairdneri)的生物测定中,注射前用抗血清预处理可完全消除远钙素的生物学效应。然而,该抗血清本身在中和内源性远钙素方面无效。对鲑鱼CS poly(A)+ RNA的体外翻译产物进行免疫沉淀研究,鉴定出一种26K蛋白,其大小与cDNA测序研究预测的远钙素单体大小一致。通过蛋白质印迹分析,纯化的远钙素和粗CS提取物获得了相似的条带模式。然而,CS提取物中有一条额外的条带(32K),在纯化的激素制剂中不存在。这可能代表鲑鱼中该激素的前体形式。