Davis J S, Alila H W, West L A, Corradino R A, Hansel W
Department of Internal Medicine, University of South Florida College of Medicine, Tampa.
Mol Cell Endocrinol. 1988 Jul;58(1):43-50. doi: 10.1016/0303-7207(88)90052-4.
The present studies were conducted to determine whether the large or small bovine luteal cell was the site for the stimulatory effect of prostaglandin F2 alpha (PGF) on phospholipase C-catalyzed inositol phospholipid hydrolysis. Corpora lutea were removed from heifers during the luteal phase of the normal estrous cycle. Small luteal cells were isolated by unit-gravity sedimentation and large luteal cells were isolated by flow cytometry using a Becton Dickson FACS 440 cell sorter. PGF provoked rapid (5-30 s) and sustained (up to 30 min) increases in the levels of inositol mono-, bis-, and trisphosphates (IP, IP2, IP3, respectively) in small luteal cells. IP3 was formed more rapidly than IP2 or IP following PGF treatment. The PGF-stimulated increase in IP3 was accompanied by a transient reduction in the levels of 3H-labeled phosphatidylinositol 4,5-bisphosphate. LiCl (10 mM) enhanced inositol phosphate accumulation in response to PGF. Maximal increases in inositol phosphate accumulation were observed with 1-10 microM PGF and half-maximal increases were observed with 60 nM PGF. PGF (1-10 microM) had no effect on cAMP levels but stimulated small increases in progesterone accumulation in 30 min incubations of small luteal cells. PGF also increased the accumulation of inositol phosphates in large luteal cells. The increases were apparent within 5 min of incubation (the earliest time examined) and further increases were observed in incubations lasting 30 min. PGF had no significant effect on cAMP or progesterone in 30 min incubations of large cells.(ABSTRACT TRUNCATED AT 250 WORDS)
进行本研究以确定大或小的牛黄体细胞是否是前列腺素F2α(PGF)对磷脂酶C催化的肌醇磷脂水解产生刺激作用的位点。在正常发情周期的黄体期从小母牛身上取出黄体。通过单位重力沉降分离小黄体细胞,使用Becton Dickson FACS 440细胞分选仪通过流式细胞术分离大黄体细胞。PGF可使小黄体细胞中的肌醇单磷酸、双磷酸和三磷酸(分别为IP、IP2、IP3)水平迅速(5 - 30秒)且持续(长达30分钟)升高。PGF处理后,IP3的形成比IP2或IP更快。PGF刺激的IP3升高伴随着3H标记的磷脂酰肌醇4,5 - 二磷酸水平的短暂降低。LiCl(10 mM)增强了对PGF的肌醇磷酸积累反应。在1 - 10 μM PGF时观察到肌醇磷酸积累的最大增加,在60 nM PGF时观察到半最大增加。PGF(1 - 10 μM)对cAMP水平无影响,但在小黄体细胞30分钟孵育中刺激孕酮积累有小幅增加。PGF也增加了大黄体细胞中肌醇磷酸的积累。孵育5分钟内(最早检测时间)增加明显,在持续30分钟的孵育中进一步增加。在大细胞30分钟孵育中,PGF对cAMP或孕酮无显著影响。(摘要截断于250字)