Serum growth hormone binding protein and hepatic GH binding sites in the Lewis dwarf rat: effects of IGF-I and GH.

Growth regulation Pub Date : 1994-12-01
R Barnard, J Mulcahy, J García-Aragón, B Wyse, P C Owens, S W Rowlinson, F Talamantes, W R Baumbach, M J Waters
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Abstract

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)

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Lewis侏儒大鼠血清生长激素结合蛋白和肝脏生长激素结合位点:igf - 1和生长激素的影响。
采用合成的鼠生长激素结合蛋白(GHBP)亲水性羧基末端序列对应的肽段作为标准,以单克隆抗体(MAb 4.3)作为一抗,建立了大鼠生长激素结合蛋白(GHBP)的放射免疫测定法(RIA)。用这种方法获得的正常大鼠GHBP浓度值与凝胶过滤和ELISA方法获得的值比较有利。11周龄正常雄性大鼠GHBP浓度(680 +/- 30 ng/ml, SEM, n = 9)显著低于正常雌性大鼠GHBP浓度(943 +/- 47 ng/ml, SEM, n = 25)。11周龄侏儒雄性大鼠GHBP浓度为423 +/- 35 ng/ml (n = 8);矮个子大鼠GHBP浓度不随年龄增长而增加,矮个子大鼠GHBP浓度随年龄增长而增加(542 +/- 32,P < 0.05, n = 9),正常雄性GHBP浓度为680 +/- 30,P < 0.001, n = 9)。不受内源性生长激素干扰的RIA的获得,将有助于进一步研究大鼠GHBP的激素和营养调节。采用该方法研究igf - 1输注(240微克/天,连续1周)和生长激素注射(65微克/100 g体重,每日2次,连续1周和4周)对11周龄Lewis侏儒大鼠血清GHBP浓度的影响。在同一动物的去饱和膜中也测量了肝生长激素结合位点。(摘要删节250字)
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