Discussion paper: induction by progesterone and a "maturation-promoting factor" of soluble proteins in Xenopus laevis oocytes in vitro

Abstract

In vitro incubation of isolated X. laevis oocytes with progesterone induces the specific labeling of proteins during maturation. They are detected as several discrete peaks with a double-labeling technique that involves the injection of [3H] leucine into hormone-treated and [14C] leucine into control cells. The peaks are separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Identical proteins are obtained with two other steroids that provoke maturation and with p-chloromercuribenzoate. The same progesterone-induced proteins are observed even if oocytes have been treated with actinomycin D or ethidium bromide or have been enucleated. Cycloheximide suppresses the labeling of proteins that are induced by progesterone, if administered either at the time of radioactive amino acid exposure or after prelabeling the oocytes to determine whether the hormone-induced peaks are due to compartmental changes of the proteins. A cytoplasmic fraction, obtained from maturing oocytes before GVBD, promotes maturation when injected into resting oocytes and therefore contains an MPF. After this injection, GVBD occurs earlier than during incubation with progesterone and is accompanied by the formation of the same proteins and of MPF, both suppressed by cycloheximide. The possibility that MPF is an induced protein is then discussed. It has been also found that there is a decrease in membrane permeability, as evidenced by decreased leucine uptake into proteins of oocytes incubated in radioactive amino acids and exposed to progesterone. The same decrease occurs after injection of MPF-containing cytoplasm into oocytes.