Barnard R, Mulcahy J, García-Aragón J, Wyse B, Owens P C, Rowlinson S W, Talamantes F, Baumbach W R, Waters M J
Department of Physiology and Pharmacology, University of Queensland, St Lucia, Australia.
Growth Regul. 1994 Dec;4(4):147-54.
A radioimmunoassay (RIA) for the rat growth hormone binding protein (GHBP) was developed using a synthetic peptide (corresponding to the hydrophilic carboxyl-terminal sequence of mouse GHBP) as standard and a monoclonal antibody (MAb 4.3) reactive with this peptide as the primary antibody. The values for GHBP concentration obtained for normal rats using this assay compare favourably with those obtained by gel filtration and ELISA methods. The concentration of GHBP in normal male rats at 11 weeks of age (680 +/- 30 ng/ml, SEM, n = 9) was significantly less than the concentration in normal females (943 +/- 47 ng/ml, SEM, n = 25). In 11-week-old dwarf male rats the concentration of GHBP was 423 +/- 35 ng/ml (n = 8); less than in dwarf females (542 +/- 32, P < 0.05, n = 9) and normal males (680 +/- 30, P < 0.001, n = 9). The GHBP concentration in dwarf rats was not age-dependent, whereas in normal females the concentration of GHBP increased with age. The availability of an RIA which is not susceptible to interference by endogenous GH, will facilitate further studies on hormonal and nutritional regulation of the rat GHBP. The assay was applied to studying the effects of IGF-I infusion (240 micrograms/day for 1 week) and GH injection (65 micrograms/100 g body weight, twice daily for 1 week and 4 weeks) on the serum concentration of GHBP in 11-week-old Lewis dwarf rats. Hepatic GH binding sites were also measured in desaturated membranes from the same animals.(ABSTRACT TRUNCATED AT 250 WORDS)
我们开发了一种用于大鼠生长激素结合蛋白(GHBP)的放射免疫分析方法(RIA),该方法使用一种合成肽(对应于小鼠GHBP的亲水性羧基末端序列)作为标准品,并使用与该肽反应的单克隆抗体(MAb 4.3)作为一抗。使用该分析方法获得的正常大鼠GHBP浓度值与通过凝胶过滤和ELISA方法获得的值相比具有优势。11周龄正常雄性大鼠的GHBP浓度(680±30 ng/ml,标准误,n = 9)显著低于正常雌性大鼠(943±47 ng/ml,标准误,n = 25)。11周龄侏儒雄性大鼠的GHBP浓度为423±35 ng/ml(n = 8);低于侏儒雌性大鼠(542±32,P < 0.05,n = 9)和正常雄性大鼠(680±30,P < 0.001,n = 9)。侏儒大鼠的GHBP浓度与年龄无关,而正常雌性大鼠的GHBP浓度随年龄增加。一种不易受内源性GH干扰的RIA的可用性将有助于进一步研究大鼠GHBP的激素和营养调节。该分析方法用于研究IGF-I输注(240微克/天,持续1周)和GH注射(65微克/100克体重,每天两次,持续1周和4周)对11周龄Lewis侏儒大鼠血清GHBP浓度的影响。还测量了同一动物去饱和膜中的肝脏GH结合位点。(摘要截断于250字)