Method for the preparation of active maturation promoting factor (MPF) from in vitro matured oocytes of Xenopus laevis.
A method for the large scale extraction of Maturation Promoting Factor (MPF) from in vitro matured oocytes of Xenopus laevis is described. MPF has been previously described only as a component(s) of hormone-matured cytoplasm within amphibian oocytes (or eggs) which is able to induce the reinitiation of the meiotic process from late diplotene stage until second metaphase arrest, when microinjected into diplotene arrested (fully grown) recipient oocytes. Standard biochemical methods for the extraction and purification of this factor(s) haven been unsuccessful due to its extreme instability and sensitivity to dilution. The procedure is dependent upon the inclusion of sodium fluoride (NaF) in the extraction medium with its effect presumably due to its ability to inhibit phosphorprotein phosphatases. The successful preservation of MPF activity described in this report permits further attempts to be made to isolate and characterize this, to date, elusive cytoplasmic factor, which plays a key role in the complex cellular processes involved in the hormone-dependent differentiation of an oocyte into an egg.
D000818:Animals; D005260:Female; D006133:Growth Substances / Q000302:isolation & purification*; D009865:Oocytes / Q000032:analysis*; D010063:Ovum / Q000032:analysis*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D012969:Sodium Fluoride; D014981:Xenopus
D006133:Growth Substances; D012969:Sodium Fluoride; D010749:Phosphoprotein Phosphatases