title: Characterization of protein kinase C-delta in mouse oocytes throughout meiotic maturation and following egg activation.
abstract: Changes in protein kinase C (PKC) activity influence the progression of meiosis; however, the specific function of the various PKC isoforms in female gametes is not known. In the current study, the protein expression and subcellular distribution profile of PKC-delta (PKC-delta), a novel isoform of the PKC family, was determined in mouse oocytes undergoing meiotic maturation and following egg activation. The full-length protein was observed as a doublet (76 and 78 kDa) on Western blot analysis. A smaller (47 kDa) carboxyl-terminal fragment, presumably the truncated catalytic domain of PKC-delta, was also strongly expressed. Both the full-length protein and the catalytic fragment became phosphorylated coincident with the resumption of meiosis and remained phosphorylated throughout metaphase II (MII) arrest. Immunofluorescence staining showed PKC-delta distributed diffusely throughout the cytoplasm of oocytes during maturation and associated with the spindle apparatus during the first meiotic division. Discrete foci of the protein also localized with the chromosomes in some mature eggs. Following the completion of meiosis, PKC-delta became dephosphorylated within 2 h of in vitro fertilization or parthenogenetic activation. The protein also accumulated in the nuclei of early embryos and was phosphorylated during M-phase of the initial mitotic cleavage division. By the two-cell stage, expression of the truncated catalytic fragment was minimal. These data demonstrate that the subcellular distribution and posttranslational modification of PKC-delta is cell cycle dependent, suggesting that its activity and/or function likely vary with the progression of meiosis and egg activation.
authors.affiliation: The Jackson Laboratory, Bar Harbor, Maine 04609, USA. viveiros@vet.upenn.edu
authors.forename: Maria M
authors.initials: MM
authors.lastname: Viveiros
authors.forename: Marilyn
authors.initials: M
authors.lastname: O'Brien
authors.forename: Karen
authors.initials: K
authors.lastname: Wigglesworth
authors.forename: John J
authors.initials: JJ
authors.lastname: Eppig
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002469:Cell Separation; D002478:Cells, Cultured; D005260:Female; D005307:Fertilization in Vitro; D005455:Fluorescent Antibody Technique; D008540:Meiosis / Q000502:physiology*; D051379:Mice; D009865:Oocytes / Q000201:enzymology*; D010063:Ovum / Q000502:physiology*; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry; D010766:Phosphorylation; D011247:Pregnancy; D014176:Protein Biosynthesis; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism*; D051745:Protein Kinase C-delta; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: C497259:Prkcd protein, mouse; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D010744:Phosphoric Monoester Hydrolases