Maller J L, Butcher F R, Krebs E G
J Biol Chem. 1979 Feb 10;254(3):579-82.
Progesterone treatment of Xenopus oocytes in vitro causes progression through meiotic cell division. The role of altered intracellular levels of cAMP on the initiation of meiotic cell division has been studied. Basal levels of cAMP averaged 1.5 pmol in oocytes from eight females, and exposure to progesterone caused a rapid drop in cAMP to about 40 to 60% of basal. Half-maximal decreases occurred within 15 to 60 s, and cAMP returned to near basal values by 20 min after progesterone. Theophylline inhibition of progesterone-induced cell division was characterized by a small increase in basal levels of cAMP and a reduced drop in cAMP due to the hormone. Cholera toxin, an activator of adenylate cyclase, was found to be a potent inhibitor of progesterone-induced meiosis, with half-maximal inhibition at 8 times 10(-12) M. In addition, the purified A subunit of cholera toxin was an effective inhibitor of progesterone action when microinjected into oocytes, with half-maximal inhibition occurring at an approximate internal concentration of 1 X 10(-7) M. Cholera toxin alone increased cAMP levels by 20%, but upon addition of progesterone, the level increased transiently to 200% of basal, indicating that the inhibition was due to elevated levels of cAMP. The results support a model in which the initiation of meiotic cell division is regulated by cAMP and protein phosphorylation.
体外对非洲爪蟾卵母细胞进行孕酮处理会导致减数分裂细胞分裂进程的推进。人们已经研究了细胞内cAMP水平的改变对减数分裂细胞分裂起始的作用。来自8只雌性的卵母细胞中cAMP的基础水平平均为1.5皮摩尔,暴露于孕酮会导致cAMP迅速下降至基础水平的约40%至60%。半数最大降幅在15至60秒内出现,孕酮处理后20分钟cAMP恢复至接近基础值。茶碱对孕酮诱导的细胞分裂的抑制作用表现为cAMP基础水平略有升高以及因激素导致的cAMP降幅减小。霍乱毒素是一种腺苷酸环化酶激活剂,被发现是孕酮诱导的减数分裂的有效抑制剂,半数最大抑制浓度为8×10⁻¹² M。此外,纯化的霍乱毒素A亚基微注射到卵母细胞中时是孕酮作用的有效抑制剂,半数最大抑制发生在大约1×10⁻⁷ M的内部浓度。单独的霍乱毒素使cAMP水平升高20%,但加入孕酮后,水平会短暂升高至基础水平的200%,这表明抑制是由于cAMP水平升高所致。这些结果支持了一个模型,即减数分裂细胞分裂的起始受cAMP和蛋白质磷酸化的调节。