Sheridan M A, Eilertson C D, Plisetskaya E M
Department of Zoology, North Dakota State University, Fargo 58105.
Gen Comp Endocrinol. 1991 Mar;81(3):365-72. doi: 10.1016/0016-6480(91)90163-z.
A specific and sensitive radioimmunoassay (RIA) for the measurement of plasma levels of somatostatin-25 (SS-25) in salmon was developed using antisera raised against coho salmon (Oncorhynchus kisutch) SS-25. Somatostatin-25 was iodinated by the chloramine-T method and repurified on Sephadex G-25. The RIA was performed using a double antibody (goat anti-rabbit gammaglobulin as second antibody) method under disequilibrium conditions. Plasma from several salmonids (coho, chinook, rainbow trout, brook trout, arctic char, lake trout, and whitefish) as well as plasma from some nonsalmonids (sucker, bluegill) cross-reacted with the antisera; serial dilutions of plasma from rainbow trout, brook trout, chinook salmon, and coho salmon were parallel to the SS-25 standard curve. Plasma from catfish showed negligible cross-reactivity. None of the mammalian somatostatins (somatostatin-14, somatostatin-28). U II, or other pancreatic hormones (insulin, glucagon) tested showed significant cross-reactivity with the antibody in the assay system. The lowest detectable level of SS-25 was 5 pg/tube; especially reproducible results were obtained in the range of 0.15-1.20 ng/ml, which appears to be the normal range of SS-25 circulating in the plasma of salmonids. Intra- and interassay coefficients of variation were 5.7 and 12.6%, respectively. Injection of glucose into chinook salmon resulted in an elevation of plasma SS-25 titers within 30 min and was coincident with hyperglycemia.
利用针对银大麻哈鱼(Oncorhynchus kisutch)生长抑素 - 25(SS - 25)制备的抗血清,开发了一种用于测量鲑鱼血浆中SS - 25水平的特异性和灵敏性放射免疫分析(RIA)方法。生长抑素 - 25通过氯胺 - T法进行碘化,并在葡聚糖凝胶G - 25上进行再纯化。RIA采用双抗体(山羊抗兔γ球蛋白作为第二抗体)法在非平衡条件下进行。几种鲑科鱼类(银大麻哈鱼、大鳞大麻哈鱼、虹鳟、溪鳟、北极红点鲑、湖鳟和白鲑)的血浆以及一些非鲑科鱼类(吸盘鱼、蓝鳃太阳鱼)的血浆与抗血清发生交叉反应;虹鳟、溪鳟、大鳞大麻哈鱼和银大麻哈鱼血浆的系列稀释与SS - 25标准曲线平行。鲶鱼的血浆显示出可忽略不计的交叉反应性。所测试的哺乳动物生长抑素(生长抑素 - 14、生长抑素 - 28)、U II或其他胰腺激素(胰岛素、胰高血糖素)在该分析系统中均未显示与抗体有显著交叉反应。SS - 25的最低可检测水平为5 pg/管;在0.15 - 1.20 ng/ml范围内获得了特别可重复的结果,这似乎是鲑科鱼类血浆中循环的SS - 25的正常范围。批内和批间变异系数分别为5.7%和12.6%。向大鳞大麻哈鱼注射葡萄糖导致血浆SS - 25滴度在30分钟内升高,且与高血糖症同时出现。