Rani C S, Nordenström K, Norjavaara E, Ahrén K
Biol Reprod. 1983 Jun;28(5):1021-31. doi: 10.1095/biolreprod28.5.1021.
Factors responsible for development of catecholamine (CA) responsiveness in granulosa cells (Gc) of preovulatory follicles from immature rats injected with 10 IU of pregnant mare's serum gonadotropin (PMSG) on Day 26 were studied. CA did not stimulate cyclic AMP (cAMP) production in whole follicles isolated before (morning) or after (evening) the preovulatory gonadotropin surge on Day 28, while newly formed corpora lutea found on Day 29 responded to CA. Gc from the preovulatory follicles did not respond to CA when tested immediately after isolation. Gc were cultured for various periods in Eagle's MEM without serum and subsequently tested for a possible stimulation of cAMP and steroidogenic responses by CA. In Gc from follicles isolated in the morning (AM-Gc) and cultured for 12 or 24 h, no response to CA was found, while the Gc isolated in the evening (PM-Gc) and cultured similarly for 12 h showed a marked response to CA both in stimulation of cAMP and progesterone production. By a total or partial elimination of the gonadotropin surge, using pentobarbital in combination with luteinizing hormone (LH) or using specific antisera to LH or follicle-stimulating hormone (FSH), it was found that previous exposure to LH was necessary for the PM-Gc to develop CA responsiveness during culture. Further, it was possible to induce CA responsiveness in AM-Gc by treatment of rats with LH for a short period in vivo followed by a period of culture. The appearance of CA responsiveness in PM-Gc cultured for 12 h was abolished when cycloheximide (5 micrograms/ml) was present during culture. It appears that the following three conditions have to be satisfied for the isolated Gc to develop CA responsiveness: 1) exposure to LH in vivo, 2) culture for a short period, and 3) an active protein synthesis. It is concluded that under physiological conditions the process of luteinization is associated with acquisition of responsiveness to CA and this process depends on the LH component of the preovulatory gonadotropin surge.
研究了在第26天注射10国际单位孕马血清促性腺激素（PMSG）的未成熟大鼠排卵前卵泡颗粒细胞（Gc）中儿茶酚胺（CA）反应性发展的相关因素。在第28天排卵前促性腺激素峰出现之前（上午）或之后（晚上）分离的完整卵泡中，CA均未刺激环磷酸腺苷（cAMP）的产生，而在第29天发现的新形成的黄体对CA有反应。排卵前卵泡分离后立即检测时，其Gc对CA无反应。将Gc在无血清的伊格尔氏基本培养基（Eagle's MEM）中培养不同时间，随后检测CA对cAMP和类固醇生成反应的可能刺激作用。在上午分离的卵泡的Gc（AM-Gc）中培养12或24小时，未发现对CA有反应，而在晚上分离的Gc（PM-Gc）中进行类似的12小时培养时，在刺激cAMP和孕酮产生方面均显示出对CA的明显反应。通过使用戊巴比妥与促黄体生成素（LH）联合或使用LH或促卵泡生成素（FSH）的特异性抗血清完全或部分消除促性腺激素峰，发现PM-Gc在培养过程中产生CA反应性之前必须预先接触LH。此外，通过在体内用LH对大鼠进行短期处理，然后进行一段时间的培养，可以诱导AM-Gc产生CA反应性。当在培养过程中存在环己酰亚胺（5微克/毫升）时，培养12小时的PM-Gc中CA反应性的出现被消除。似乎分离的Gc要产生CA反应性必须满足以下三个条件：1）体内接触LH；2）短期培养；3）活跃的蛋白质合成。得出的结论是，在生理条件下，黄体化过程与对CA反应性的获得有关，并且这个过程取决于排卵前促性腺激素峰中的LH成分。
