Progesterone release of lipid second messengers at the amphibian oocyte plasma membrane: role of ceramide in initiating the G2/M transition.

Treatment with either sphingomyelinase (SMase), or soluble forms of ceramide, has been reported to induce meiosis in oocytes from Xenopus laevis, a species which can breed throughout most of the year. In this paper the sphingomyelin-derived second messenger, ceramide, is compared with progesterone for its ability to induce meiosis in oocytes from the seasonal breeder, Rana pipiens. Serum gonadotropin levels normally rise as Rana females emerge from hibernation in the spring, stimulating follicular synthesis of progesterone and subsequent ovulation. Injection of gonadotropins can induce earlier meiosis and ovulation, effective from the previous October through the following spring. During the same period, defolliculated oocytes respond to exogenous progesterone by meiosis, as indicated by nuclear breakdown. We find that in the spring, treatment of defolliculated Rana oocytes with exogenous C2- or C8-ceramide or SMase did induce meiosis, but not during the fall or winter. A 50% response was seen by late April and a 100% response by early May. Exposure of [3H]palmitate-labeled Rana oocytes to either exogenous progesterone or to SMase produced a rapid and comparable release of intracellular [3H]ceramide within 1-2 min in fall, winter or spring. Our results from this and from previous experiments indicate that increased ceramide is not the initiating event in meiotic induction in Rana, but is associated with a subsequent pathway which depends upon a threshold level of progesterone.