
doi: 10.1530/rep.1.00824
pmid: 16816331
Abstract Reinitiation of meiosis in meiotically competent, fully grown mammalian oocytes is governed by a fall in intraoocyte cAMP concentrations and the subsequent inactivation of protein kinase A (PKA). A similar reduction in intraoocyte cAMP concentrations in growing, meiotically incompetent rat oocytes not leading to resumption of meiosis, questions the involvement of PKA in the regulation of meiosis at this early stage of oocyte development. We examined the possibility of whether PKA activity maintains growing oocytes in meiotic arrest and further explored the mode of activation of PKA under conditions of relatively low cAMP concentrations. Our experiment demonstrated that inactivation of PKA stimulates growing rat oocytes to resume meiosis, and elevates the activity of their maturation-promoting factor (MPF). We also found that the expressions of type I and type II regulatory subunits (RI and RII) of PKA are higher in growing and fully grown oocytes, respectively. In addition, we revealed that the common 1:1 ratio between the regulatory (R) and catalytic (C) subunits of PKA is apparently not abrogated and, in accordance PKA activity in growing oocyte - cell extract is fully dependent on cAMP. Finally, we identified in growing oocytes, the A kinase anchoring protein (AKAP) 140, which was previously depicted in fully grown oocytes. We conclude that an active PKA prevents growing oocytes from resuming meiosis. Our findings further suggest that relatively high abundance of the PKAI isoform and/or its subcellular compartmentalization, through interaction with AKAP140, could possibly account for the high basal PKA activity at relatively low intraoocyte cAMP concentrations.
Cyclic AMP-Dependent Protein Kinase RIalpha Subunit, Blotting, Western, Maturation-Promoting Factor, A Kinase Anchor Proteins, Fluorescent Antibody Technique, Cyclic AMP-Dependent Protein Kinase Type II, Isoquinolines, Cyclic AMP-Dependent Protein Kinases, Enzyme Activation, Meiosis, Oogenesis, 1-Methyl-3-isobutylxanthine, Cyclic AMP-Dependent Protein Kinase RIIalpha Subunit, Cyclic AMP, Oocytes, Animals, Female, Enzyme Inhibitors, Cells, Cultured, Adaptor Proteins, Signal Transducing
Cyclic AMP-Dependent Protein Kinase RIalpha Subunit, Blotting, Western, Maturation-Promoting Factor, A Kinase Anchor Proteins, Fluorescent Antibody Technique, Cyclic AMP-Dependent Protein Kinase Type II, Isoquinolines, Cyclic AMP-Dependent Protein Kinases, Enzyme Activation, Meiosis, Oogenesis, 1-Methyl-3-isobutylxanthine, Cyclic AMP-Dependent Protein Kinase RIIalpha Subunit, Cyclic AMP, Oocytes, Animals, Female, Enzyme Inhibitors, Cells, Cultured, Adaptor Proteins, Signal Transducing
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