Mitogen-activated protein kinase activity and microtubule organisation are altered by protein synthesis inhibition in maturing porcine oocytes.
Previously we have shown that mitogen-activated protein (MAP) kinase activity abruptly increases at the first metaphase (M1) and remains significantly higher than that at the germinal vesicle (GV) stage until the second metaphase (M2) in porcine oocytes cultured in vitro. The present paper describes how the mechanism of the blockage of meiotic maturation by protein synthesis inhibition involves MAP kinase regulation. Cycloheximide arrested both germinal vesicle breakdown (GVBD) and the normal transition from M1 to M2. MAP kinase activation was also reduced in these maturation-inhibited oocytes. By using immunofluorescence microscopy with the monoclonal antibody raised against rat alpha-tubulin, we showed that cycloheximide caused morphological abnormality in a spindle at M1, but not at M2. All these results indicate that in porcine oocytes: (1) GV blockage by protein synthesis inhibition involves the suppression of both histone H1 kinase and MAP kinase activation, (2) during the transition from M1 to M2, maintenance of a normal metaphasic spindle and high MAP kinase activity require protein synthesis, and (3) once the M2 cytoskeletal structures have been completed, and/or after the 'critical period', cytostatic factor activity is independent of protein synthesis.
Inoue
M
M
Department of Reproductive and Developmental Biology, University of Tokyo, Japan.
Naito
K
K
Nakayama
T
T
Sato
E
E
D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000032:analysis / Q000378:metabolism*; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D003513:Cycloheximide / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D008540:Meiosis / Q000502:physiology; D008677:Metaphase / Q000502:physiology; D008870:Microtubules / Q000502:physiology*; D009865:Oocytes / Q000166:cytology* / Q000201:enzymology; D006656:Protamine Kinase / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D013552:Swine
D011500:Protein Synthesis Inhibitors; D003513:Cycloheximide; D006656:Protamine Kinase; D017871:Calcium-Calmodulin-Dependent Protein Kinases