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用于奇努克鲑(Oncorhynchus tshawytscha)生长激素的高灵敏度放射免疫测定法的开发与验证。

Development and validation of a highly sensitive radioimmunoassay for chinook salmon (Oncorhynchus tshawytscha) growth hormone.

作者信息

Le Bail P Y, Sumpter J P, Carragher J F, Mourot B, Niu P D, Weil C

机构信息

Laboratoire de Physiologie des Poissons, INRA, Rennes, France.

出版信息

Gen Comp Endocrinol. 1991 Jul;83(1):75-85. doi: 10.1016/0016-6480(91)90107-h.

Abstract

This study describes the development of a highly specific and very sensitive radioimmunoassay for salmonid growth hormone. Antiserum raised against chinook (Oncorhynchus tshawytscha) GH2, which did not recognize 125I-sPRL and 125I-sGTH (at 1:1000 initial dilution), was able to inhibit growth when injected into rainbow trout (Oncorhynchus mykiss). 125I-sGH2, used as tracer, was not recognized by anti-sGTH or by anti-sPRL. Mammalian GH and ACTH and salmonid GTH, TSH, and PRL did not cross-react in the sGH assay. The inhibition curves for pituitary extracts and plasma from salmonids were parallel to the salmon GH standard, whereas those from carp, tilapia, and catfish showed no significant cross reactivity. The RIA ED90 and ED50 values were 0.2 and 1.5 ng/ml, respectively. Using this RIA for measuring GH release by cultured pituitary cell we observed a strong inhibiting effect of SRIF (10(-6) M) and a stimulatory effect of hGRF (10(-6) M). This RIA allowed us also to detect daily fluctuations in the plasma GH concentration in cannulated rainbow trout.

摘要

本研究描述了一种针对鲑鱼生长激素的高特异性和高灵敏度放射免疫分析方法的开发。用奇努克鲑(Oncorhynchus tshawytscha)GH2制备的抗血清,在初始稀释度为1:1000时不识别125I-sPRL和125I-sGTH,将其注射到虹鳟(Oncorhynchus mykiss)体内时能够抑制生长。用作示踪剂的125I-sGH2不被抗sGTH或抗sPRL识别。哺乳动物生长激素和促肾上腺皮质激素以及鲑鱼促性腺激素、促甲状腺激素和催乳素在sGH分析中不发生交叉反应。鲑鱼垂体提取物和血浆的抑制曲线与鲑鱼生长激素标准曲线平行,而鲤鱼、罗非鱼和鲶鱼的抑制曲线未显示出明显交叉反应。放射免疫分析的ED90和ED50值分别为0.2和1.5 ng/ml。使用这种放射免疫分析法测量培养垂体细胞释放的生长激素,我们观察到生长抑素(10(-6) M)有强烈的抑制作用,而人促生长激素释放因子(10(-6) M)有刺激作用。这种放射免疫分析法还使我们能够检测插管虹鳟血浆中生长激素浓度的每日波动情况。

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