Endocytosis in the amphibian oocyte. Effect of insulin and progesterone on membrane and fluid internalization during the meiotic divisions.

Endocytosis has been studied in the denuded Rana pipiens oocyte using 3H-labeled inulin. Internalization of labeled inulin is linear after the first 10-15 min and uptake into the cytoplasm is temperature-dependent and is blocked by 15 microM cyanide. Uptake occurs without hydrolysis of the inulin and varies exponentially with the concentration of inulin in the medium. Based on specific activity of the medium and inulin uptake into the cytoplasm, it is estimated that a fluid volume of about 20-25 nl is internalized per oocyte per hour. This fluid phase uptake corresponds to a half-time of about 35 h for turnover of the oocyte fluid phase. An estimate of membrane area based on endocytotic vesicle size from electron micrographs suggests that the entire oocyte plasma membrane recycles several times an hour. A fraction (15-20%) of the inulin taken up is associated with the plasma-vitelline membrane complex and uptake into the membrane complex parallels uptake into the cytoplasm. Insulin (a meiotic agonist) concentrations that induce plasma membrane hyperpolarization over the first h also stimulate [3H]inulin uptake into both the oocyte cytoplasm and membrane complex over the same time period. Progesterone (the physiological inducer) has no effect on inulin uptake during the first hour, but by 16-17 h after exposure to progesterone, inulin uptake is significantly enhanced. These results suggest that hormones such as insulin and progesterone may regulate membrane permeability by a programmed internalization and possible recycling of the plasma membrane components.