Okadaic acid accelerates germinal vesicle breakdown and overcomes cycloheximide- and 6-dimethylaminopurine block in cattle and pig oocytes.
Pig and cattle oocytes, when released from the follicle, spontaneously resume first meiotic division within 20 or 8 hr, respectively. In oocytes of both species, the activity of histone H1 kinase increases during maturation, exhibiting a maximum in metaphase I. Treatment of these oocytes with okadaic acid results in acceleration of germinal vesicle breakdown (GVBD) and of histone H1 kinase activation. This effect is more important in pig oocytes, in which the acceleration rises for 6 hr, as compared to 2 hr in cattle. Moreover, under these conditions, H1 kinase activity measured after 12 hr of culture appears higher than that observed in control metaphase I oocytes. When added to prophase oocytes, both cycloheximide and 6-DMAP (6-dimethylaminopurine) block GVBD and histone H1 kinase activation. Okadaic acid, at a concentration of 2.5 microM, is able to release the inhibitory effect exerted by cycloheximide on histone H1 kinase activity; however, GVBD occurred only in two-thirds of pig and one-quarter of cattle oocytes after 20 hr of culture. In addition, okadaic acid fully reverses the effect of 6-DMAP on H1 kinase activity and on GVBD in both species. The opposite effects of 6-DMAP and okadaic acid on MPF activation are discussed, as well as the nature of the protein, which has to be synthesized during the first meiotic division and may be involved in the MPF activation cascade.
Kalous
J
J
Department of Genetics, Czechoslovak Academy of Sciences, Libechov.
Kubelka
M
M
Rimkevicova
Z
Z
Guerrier
P
P
Motlik
J
J
D000225:Adenine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D003513:Cycloheximide / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects*; D004988:Ethers, Cyclic / Q000494:pharmacology*; D005260:Female; D016200:Maturation-Promoting Factor / Q000187:drug effects; D019319:Okadaic Acid; D009865:Oocytes / Q000187:drug effects* / Q000201:enzymology; D006656:Protamine Kinase / Q000096:biosynthesis*; D013552:Swine
D004988:Ethers, Cyclic; D019319:Okadaic Acid; C001020:N(6),N(6)-dimethyladenine; D003513:Cycloheximide; D006656:Protamine Kinase; D016200:Maturation-Promoting Factor; D000225:Adenine