Inhibition of protein kinases by 6-dimethylaminopurine accelerates the transition to interphase in activated mouse oocytes.
Mouse oocyte activation is followed by a peculiar period during which the interphase network of microtubules does not form and the chromosomes remain condensed despite the inactivation of MPF. To evaluate the role of protein phosphorylation during this period, we studied the effects of the protein kinase inhibitor 6-dimethylaminopurine (6-DMAP) on fertilization and/or parthenogenetic activation of metaphase II-arrested mouse oocytes. 6-DMAP by itself does not induce the inactivation of histone H1 kinase in metaphase II-arrested oocytes, and does not influence the dynamics of histone H1 kinase inactivation during oocyte activation. However, 6-DMAP inhibits protein phosphorylation after oocyte activation. In addition, the phosphorylated form of some proteins disappear earlier in oocytes activated in the presence of 6-DMAP than in the activated control oocytes. This is correlated with the acceleration of some post-fertilization morphological events, such as sperm chromatin decondensation and its transient recondensation, formation of the interphase network of microtubules and pronuclear formation. In addition, numerous abnormalities could be observed: (1) the spindle rotation and polar body extrusion are inhibited; (2) the exchange of protamines into histones seems to be impaired, as judged by the morphology of DNA fibrils by electron microscopy; (3) the formation of a new nuclear envelope around the sperm chromatin proceeds prematurely, while recondensation is not yet completed. These observations suggest that the 6-DMAP-sensitive kinase(s) is (are) involved in the control of post-fertilization events such as the formation of the interphase network of microtubules, the remodelling of sperm chromatin and pronucleus formation.
Szöllösi
M S
MS
INRA, Unité de Biologie de la Fécondation, Jouy-en-Josas, France.
Kubiak
J Z
JZ
Debey
P
P
de Pennart
H
H
Szöllösi
D
D
Maro
B
B
D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects; D002843:Chromatin / Q000187:drug effects*; D004527:Egg Proteins / Q000187:drug effects / Q000378:metabolism; D005260:Female; D007399:Interphase / Q000187:drug effects*; D008297:Male; D000090204:Mesothelin; D051379:Mice; D008870:Microtubules / Q000187:drug effects*; D008970:Molecular Weight; D009865:Oocytes / Q000166:cytology / Q000187:drug effects*; D010766:Phosphorylation; D006656:Protamine Kinase / Q000037:antagonists & inhibitors; D047428:Protein Kinase Inhibitors; D013094:Spermatozoa / Q000187:drug effects; D013997:Time Factors
D002843:Chromatin; D004527:Egg Proteins; C000715528:Msln protein, mouse; D047428:Protein Kinase Inhibitors; C001020:N(6),N(6)-dimethyladenine; D006656:Protamine Kinase; D000090204:Mesothelin; D000225:Adenine