Joensuu T, Tuohimaa P, Vilja P
Department of Biomedical Sciences, University of Tampere, Finland.
J Endocrinol. 1991 Aug;130(2):191-7. doi: 10.1677/joe.0.1300191.
This study describes sensitive immunoenzymometric assays (IEMAs) for chicken avidin and ovalbumin, markers of cytodifferentiation and action of progesterone and oestrogen in the oviduct magnum mucosa. The determination range was 0.5-100 ng/ml and the detection limit 0.1 ng/ml in both IEMAs. The intra- and interassay coefficients of variation, measured from chicken tissue supernatants, averaged below 6 and 10% respectively. IEMAs correlated well with the radioimmunoassays for avidin and ovalbumin previously developed in our laboratory, and with the widely used [14C]biotin-binding method for avidin. Using an IEMA, we found avidin induction with low concentrations of progesterone in the differentiated oviduct of oestrogen-pretreated chicks. The induction has not been detected previously by less sensitive methods. Avidin was induced by all given doses of progesterone (0.2-200 mg/kg in vivo for 24 h after a short oestrogen treatment), the response being dose-dependent at doses of 0.2-20 mg progesterone/kg body weight, the maximum avidin production being about 70 micrograms/g tissue. Ovalbumin was induced at doses of 2-200 mg progesterone/kg body weight without variations in the responses, being about 35 mg/g. The mean content of avidin in the oviduct of laying hens was 58.1 micrograms/g, and of ovalbumin 74.9 mg/g. Minimal traces of avidin and ovalbumin were found in the oviduct after hatching (0.3 and 5 micrograms/g respectively); however, progesterone did not have an effect on this expression. Sensitivity, rapidity and practicability, together with non-radioactivity, are the main advantages of the present IEMAs for chicken avidin and ovalbumin.
本研究描述了用于检测鸡抗生物素蛋白和卵清蛋白的灵敏免疫酶量分析法(IEMAs),这两种蛋白是输卵管峡部黏膜中细胞分化以及孕酮和雌激素作用的标志物。两种IEMAs的测定范围均为0.5 - 100 ng/ml,检测限均为0.1 ng/ml。从鸡组织上清液测得的批内和批间变异系数平均分别低于6%和10%。IEMAs与我们实验室先前开发的抗生物素蛋白和卵清蛋白放射免疫分析法以及广泛使用的抗生物素蛋白[14C]生物素结合法相关性良好。使用IEMA,我们发现在雌激素预处理雏鸡的分化输卵管中,低浓度孕酮可诱导抗生物素蛋白的产生。以前用不太灵敏的方法未检测到这种诱导作用。所有给定剂量的孕酮(短期雌激素处理后,体内0.2 - 200 mg/kg,处理24小时)均可诱导抗生物素蛋白的产生,在0.2 - 20 mg孕酮/kg体重剂量下,反应呈剂量依赖性,抗生物素蛋白的最大产量约为70微克/克组织。在2 - 200 mg孕酮/kg体重剂量下可诱导卵清蛋白的产生,反应无变化,约为35毫克/克。产蛋母鸡输卵管中抗生物素蛋白的平均含量为58.1微克/克,卵清蛋白为74.9毫克/克。孵化后在输卵管中发现极微量的抗生物素蛋白和卵清蛋白(分别为0.3和5微克/克);然而,孕酮对此表达没有影响。灵敏度、快速性和实用性,以及无放射性,是目前用于检测鸡抗生物素蛋白和卵清蛋白的IEMAs的主要优点。