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促卵泡激素(FSH)体外对青春期前大鼠卵巢细胞外活性环磷酸腺苷磷酸二酯酶的刺激作用。

Stimulatory effect of FSH in vitro on the extracellularly active cyclic AMP phosphodiesterase in the prepubertal rat ovary.

作者信息

Selstam G, Rosberg S

出版信息

Acta Endocrinol (Copenh). 1976 Mar;81(3):563-73. doi: 10.1530/acta.0.0810563.

Abstract

Intact prepubertal rat ovaries were incubated with radioactively labelled adenosine 3',5'-cyclic monophosphate (cAMP) in Krebs bicarbonate buffer containing glucose. The rate of degradation of cAMP was determined by measuring the radioactivity in the medium after precipitation with Ba(OH)2 and ZnSO4. The fate of the nucleotide was followed by measuring the products in the incubation medium. Paper chromatography was used for the separation and identification of these products. It was found that cAMP was degraded to AMP, which in turn was degraded to inorganic phosphate (Pi) and adenosine. An uptake of labelled products was also observed. NIH-FSH-S9 (10 and 100 mug/ml), but not NIH-LH-B8 (0.1-100 mug/ml), increased the degradation of cAMP. Concomitantly, an increased accumulation of labelled adenosine and Pi as well as an increased uptake of labelled products were seen. Kinetic studies with low concentrations of cAMP (0.125-0.025 mumol/l) revealed an apparent Km value of 0.12 mumol/l for the phosphodiesterase (PDE) activity. FSH significantly changed the slope of the curve in the Lineweaver-Burk plot by increasing the PDE activity. The increased PDE activity in the presence of FSH is discussed in relation to earlier findings of differences in action betweeh LH and FSH on the cAMP system in the prepubertal rat ovary.

摘要

将未成熟青春期前大鼠的卵巢置于含有葡萄糖的Krebs碳酸氢盐缓冲液中,与放射性标记的3',5'-环磷酸腺苷(cAMP)一起孵育。通过用Ba(OH)2和ZnSO4沉淀后测量培养基中的放射性来确定cAMP的降解速率。通过测量孵育培养基中的产物来追踪核苷酸的去向。用纸色谱法分离和鉴定这些产物。发现cAMP降解为AMP,而AMP又降解为无机磷酸盐(Pi)和腺苷。还观察到标记产物的摄取。NIH-FSH-S9(10和100μg/ml),但不是NIH-LH-B8(0.1 - 100μg/ml),增加了cAMP的降解。同时,观察到标记腺苷和Pi的积累增加以及标记产物的摄取增加。用低浓度cAMP(0.125 - 0.025μmol/l)进行的动力学研究表明,磷酸二酯酶(PDE)活性的表观Km值为0.12μmol/l。FSH通过增加PDE活性显著改变了Lineweaver-Burk图中曲线的斜率。结合先前关于青春期前大鼠卵巢中LH和FSH对cAMP系统作用差异的研究结果,讨论了FSH存在时PDE活性增加的情况。

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