Translocation of the classic protein kinase C isoforms in porcine oocytes: implications of protein kinase C involvement in the regulation of nuclear activity and cortical granule exocytosis.
Protein kinase C (PKC) is a family of Ser/Thr protein kinases categorized into three subfamilies: classical, novel, and atypical. The subcellular localization of classical PKCalpha, -betaI, and -gamma in the process of porcine oocyte maturation, fertilization, and parthenogenetic activation and their involvement in cortical granule (CG) exocytosis were investigated. The results of Western blot showed that PKCalpha, -betaI, and -gamma were expressed in the oocytes at the germinal vesicle (GV) and metaphase II (MII) stages. Confocal microscopy revealed that the three PKC isoforms were concentrated in the GV but evenly distributed in the cytoplasm of MII eggs. PKCalpha and -gamma were translocated to the plasma membrane soon after sperm penetration. cPKCs migrated into the pronucleus in fertilized eggs. Following treatment with a PKC activator, phorbol 12-myristate 13-acetate (PMA), CGs were released and PKCalpha and -gamma were translocated to the membrane. The CG exocytosis and PKC redistribution induced by PMA could be blocked by the PKC inhibitor staurosporine. Parthenogenetic stimulation with ionophore A23187 or electrical pulse also induced cPKC translocation and CG exocytosis. Eggs injected with PKCalpha isoform-specific antibody failed to undergo CG exocytosis after PMA treatment or fertilization. The results suggest that cPKCs, especially the alpha-isotype, regulate nuclear function and CG exocytosis in porcine eggs.
Fan
Heng-Yu
HY
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China.
Tong
Chao
C
Li
Man-Yu
MY
Lian
Li
L
Chen
Da-Yuan
DY
Schatten
Heide
H
Sun
Qing-Yuan
QY
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology*; D004558:Electric Stimulation; D005089:Exocytosis / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro; D007527:Isoenzymes / Q000276:immunology / Q000378:metabolism*; D008297:Male; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000201:enzymology; D010063:Ovum; D011493:Protein Kinase C / Q000276:immunology / Q000378:metabolism*; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000906:Antibodies; D007527:Isoenzymes; C087857:protein kinase C gamma; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D013755:Tetradecanoylphorbol Acetate