Dufau M L, Hodgen G D, Goodman A L, Catt K J
Endocrinology. 1977 Jun;100(6):1557-65. doi: 10.1210/endo-100-6-1557.
The concentration of biologically active LH in rhesus monkey (Macaca mulatta) serum was measured by a highly sensitive bioassay based upon testosterone production by dispersed rat interstitial cells. The sensitivity of the in vitro bioassay was equal to or higher than that of radioimmunoassay, with detection limits of 0.1 mIU of human menopausal gonadotropin (hMG) or 10 ng of a rhesus pituitary gonadotropin preparation (LER-1909-2). Parallel dose-response curves were obtained for hMG and rhesus monkey pituitary gonadotropin. The method permits bioassy of LH in 20-100 micronl of serum from adult male monkeys, and from female monkeys during the follicular and luteal phases of the menstrual cycle. Bioactive LH concentrations could be assayed in 0.25 to 5 micronl of serum from mid-cycle, postmenopausal, and castrated female monkeys. Serum LH was undetectable in two hypophysectomized adult female monkeys and six intact immature animals, and was 13+/-6 (SD) mIU/ml in adult male monkeys. In adult females, follicular phase LH levels ranged from 17 to 169 mIU/ml, with a mean of 76+/-52 mIU/ml. The midcycle LH peak was 1738+/-742 mIU/ml and the luteal phase values ranged from 6-47 mIU/ml, with a mean of 35+/-5 mIU/ml. Serum LH concentrations ranged from 100 to 900 mIU/ml in two menopausal females, and from 590-1480 mIU/ml in castrated females. Treatment of castrated female monkeys with estrogen plus progesterone produced an initial two-fold rise in serum LH within 3 days, followed by a gradual decline to one-fourth to one-tenth of the initial levels after 10 days of treatment. Serum LH was suppressed to undetectable levels during the third week, and remained so for the duration of the 60-day treatment period. Bioactive serum LH levels were comparable to levels determined by radioimmunoassay during the follicular and luteal phases of the menstrual cycle, with increased bio-immunoratio at the midcycle peak. The concentrations of biologically active serum LH in rhesus monkeys were similar to those in the human female during the follicular and luteal phases of the menstrual cycle, and were higher at midcycle and after castration. Serum LH levels measured by the interstitial cell bioassay in the rhesus monkey showed appropriate physiological changes and responses to gonadal steroid administration. Furthermore, the bioassay did not detect the LH-like material measured by heterologous radioimmunoassay in the serum of hypophysectomized, immature and steroid-suppressed monkeys. Thus, the rat interstitial cell assay provides a sensitive and valid procedure for measurement of biologically active LH in the serum of these non-human primates.
采用一种基于分散的大鼠间质细胞产生睾酮的高灵敏度生物测定法，测定了恒河猴（猕猴）血清中生物活性促黄体生成素（LH）的浓度。体外生物测定法的灵敏度等于或高于放射免疫测定法，检测限为人绝经促性腺激素（hMG）0.1 mIU或恒河猴垂体促性腺激素制剂（LER-1909-2）10 ng。获得了hMG和恒河猴垂体促性腺激素的平行剂量反应曲线。该方法可用于测定成年雄性猴子以及处于月经周期卵泡期和黄体期的雌性猴子20 - 100微升血清中的LH。可测定处于月经周期中期、绝经后和去势雌性猴子0.25至5微升血清中的生物活性LH浓度。在两只垂体切除的成年雌性猴子和六只未成熟的完整动物中未检测到血清LH，成年雄性猴子血清LH为13±6（标准差）mIU/ml。成年雌性猴子卵泡期LH水平在17至169 mIU/ml之间，平均为76±52 mIU/ml。月经周期中期LH峰值为1738±742 mIU/ml，黄体期值在6 - 47 mIU/ml之间，平均为35±5 mIU/ml。两只绝经后雌性猴子血清LH浓度在100至900 mIU/ml之间，去势雌性猴子血清LH浓度在590 - 1480 mIU/ml之间。对去势雌性猴子用雌激素加孕激素治疗，3天内血清LH最初升高两倍，治疗10天后逐渐降至初始水平的四分之一至十分之一。在第三周血清LH被抑制到检测不到的水平，并在60天治疗期内一直保持。在月经周期的卵泡期和黄体期，生物活性血清LH水平与放射免疫测定法测定的水平相当，在月经周期中期峰值时生物免疫比增加。恒河猴血清中生物活性LH的浓度在月经周期的卵泡期和黄体期与人类女性相似，在月经周期中期和去势后更高。通过间质细胞生物测定法测定的恒河猴血清LH水平显示出适当的生理变化以及对性腺类固醇给药的反应。此外，该生物测定法未检测到垂体切除、未成熟和类固醇抑制的猴子血清中通过异源放射免疫测定法测定的LH样物质。因此，大鼠间质细胞测定法为测量这些非人灵长类动物血清中生物活性LH提供了一种灵敏且有效的方法。
