Characterization of Novel Phosphodiesterases in the Bovine Ovarian Follicle1

Abstract The phosphodiesterase (PDE) family is a group of enzymes that catalyzes the transformation of cyclic nucleotides into 5′ nucleotides. Based on rodents, the current mammalian model of PDE distribution in the ovarian follicle predicts Pde3a in the oocyte and Pde4d in the somatic cells. Using bovine as an experimental model, the present results showed that PDE3 was the predominant PDE activity in oocytes. However, cumulus cell cAMP-PDE activity was predominantly resistant to inhibition by 3-isobutyl-methylxantine, indicating PDE8 activity (60% of total PDE activity) and a minor role for PDE4 (<5%). A total of 20% of total oocyte PDE activity was also attributed to PDE8. The PDE activity measurements in mural granulosa cells from 2 to 6 mm in diameter suggest the presence of PDE4 and PDE8. In granulosa cells from follicles >10 mm, total PDE and PDE8 activities along with PDE8A protein level were increased compared with smaller follicles. The RT-PCR experiments showed that cumulus cells expressed PDE8A, PDE8B, and PDE10A. Western blot experiments showed PDE8A, PDE8B, and PDE4D proteins in mural granulosa cells and cumulus-oocyte complexes. PDE8 inhibition using dipyridamole in a dose-dependent manner increased cAMP levels in the cumulus-oocyte complexes and delayed oocyte nuclear maturation. These results are the first to demonstrate the functional presence of PDE8 in the mammalian ovarian follicle. This challenges the recently described cell-specific expression of cAMP-PDEs in the ovarian follicle and the notion that PDE4 is the predominant granulosa/cumulus cell PDE. These findings have implications for our understanding of hormonal regulation of folliculogenesis and the potential application of PDE inhibitors as novel contraceptives.

[1]  M. Conti,et al.  Oocyte maturation involves compartmentalization and opposing changes of cAMP levels in follicular somatic and germ cells: studies using selective phosphodiesterase inhibitors. , 1996, Developmental biology.

[2]  P. Epstein,et al.  Assay of cyclic nucleotide phosphodiesterase and resolution of multiple molecular forms of the enzyme. , 1979, Advances in cyclic nucleotide research.

[3]  C. Guillemette,et al.  New insight into the role of phosphodiesterase 3A in porcine oocyte maturation , 2006, BMC Developmental Biology.

[4]  M. Sirard,et al.  Effect of the Absence or Presence of Various Protein Supplements on Further Development of Bovine Oocytes During In Vitro Maturation1 , 2002, Biology of reproduction.

[5]  K. Matsushima,et al.  Molecular cloning and characterization of human PDE8B, a novel thyroid-specific isozyme of 3',5'-cyclic nucleotide phosphodiesterase. , 1998, Biochemical and biophysical research communications.

[6]  R. Cortvrindt,et al.  Effect of Phosphodiesterase Type 3 Inhibitor on Developmental Competence of Immature Mouse Oocytes In Vitro1 , 2003, Biology of reproduction.

[7]  A. Gutiérrez-Adán,et al.  Oocyte and embryo quality: effect of origin, culture conditions and gene expression patterns. , 2003, Reproduction in domestic animals = Zuchthygiene.

[8]  J. Beavo,et al.  Cloning and characterization of PDE7B, a cAMP-specific phosphodiesterase. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[9]  J. M. Garcia,et al.  Role of roscovitine and IBMX on kinetics of nuclear and cytoplasmic maturation of bovine oocytes in vitro. , 2007, Animal reproduction science.

[10]  S. Bilodeau-Goeseels,et al.  Effects of phosphodiesterase inhibitors on spontaneous nuclear maturation and cAMP concentrations in bovine oocytes. , 2003, Theriogenology.

[11]  P. Mermillod,et al.  Bovine blastocyst production in vitro after inhibition of oocyte meiotic resumption for 24 h. , 1997, Journal of reproduction and fertility.

[12]  J. Beavo,et al.  Biochemistry and physiology of cyclic nucleotide phosphodiesterases: essential components in cyclic nucleotide signaling. , 2007, Annual review of biochemistry.

[13]  M. Conti,et al.  Role of cyclic nucleotide signaling in oocyte maturation , 2002, Molecular and Cellular Endocrinology.

[14]  W. P. Kuo,et al.  Impaired growth and fertility of cAMP-specific phosphodiesterase PDE4D-deficient mice. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  P. Epstein,et al.  Phosphodiesterase 8 (PDE8) regulates chemotaxis of activated lymphocytes. , 2006, Biochemical and biophysical research communications.

[16]  J. Kotera,et al.  Molecular comparison of rat cyclic nucleotide phosphodiesterase 8 family: unique expression of PDE8B in rat brain. , 2003, Gene.

[17]  D. Armstrong,et al.  Lack of ovarian responsiveness to gonadotropic hormones in infantile rats sterilized with busulfan. , 1986, Endocrinology.

[18]  M. Wheeler,et al.  Maintenance of bovine oocytes in prophase of meiosis I by high [cAMP]i. , 1995, Journal of reproduction and fertility.

[19]  M. Conti,et al.  Phosphodiesterase 3 inhibitors suppress oocyte maturation and consequent pregnancy without affecting ovulation and cyclicity in rodents. , 1998, The Journal of clinical investigation.

[20]  J. Beavo,et al.  Cloning and characterization of a cAMP-specific cyclic nucleotide phosphodiesterase. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J. Beavo,et al.  Cloning and characterization of two splice variants of human phosphodiesterase 11A. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[22]  R. Gilchrist,et al.  Bovine Cumulus Cell-Oocyte Gap Junctional Communication During In Vitro Maturation in Response to Manipulation of Cell-Specific Cyclic Adenosine 3′,5′-Monophosophate Levels1 , 2004, Biology of reproduction.

[23]  K. Takeda,et al.  Short‐term or long‐term treatments with a phosphodiesterase‐4 (PDE4) inhibitor result in opposing agonist‐induced Ca2+ responses in endothelial cells , 2008, British journal of pharmacology.

[24]  C. Jessus,et al.  How does Xenopus oocyte acquire its competence to undergo meiotic maturation? , 2004, Biology of the cell.

[25]  R. Gilchrist,et al.  Effect of hexoses and gonadotrophin supplementation on bovine oocyte nuclear maturation during in vitro maturation in a synthetic follicle fluid medium. , 2005, Reproduction, fertility, and development.

[26]  M. Mayes,et al.  Effect of Type 3 and Type 4 Phosphodiesterase Inhibitors on the Maintenance of Bovine Oocytes in Meiotic Arrest1 , 2002, Biology of reproduction.

[27]  Y. Shu,et al.  Effects of cilostamide and forskolin on the meiotic resumption and embryonic development of immature human oocytes. , 2008, Human reproduction.

[28]  L. Zacharia,et al.  Characterization of Renal Ecto-Phosphodiesterase , 2007, Journal of Pharmacology and Experimental Therapeutics.

[29]  J. Beavo,et al.  Isolation and characterization of a dual-substrate phosphodiesterase gene family: PDE10A. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. Vassena,et al.  Role of Intracellular Cyclic Adenosine 3′,5′-Monophosphate Concentration and Oocyte-Cumulus Cells Communications on the Acquisition of the Developmental Competence During In Vitro Maturation of Bovine Oocyte1 , 2004, Biology of reproduction.

[31]  H. Callesen,et al.  In vivo development of in vitro fertilized bovine oocytes matured in vivo versus in vitro , 1987, Journal of in Vitro Fertilization and Embryo Transfer.

[32]  C. Guillemette,et al.  3'5'-cyclic adenosine monophosphate-dependent up-regulation of phosphodiesterase type 3A in porcine cumulus cells. , 2007, Endocrinology.

[33]  R. Ivell,et al.  Phosphodiesterase 4 inhibition synergizes with relaxin signaling to promote decidualization of human endometrial stromal cells. , 2004, The Journal of clinical endocrinology and metabolism.

[34]  Martin F. Laforest,et al.  Fundamental significance of specific phosphodiesterases in the control of spontaneous meiotic resumption in porcine oocytes , 2005, Molecular reproduction and development.

[35]  J. Beavo,et al.  Identification and Characterization of a Novel Family of Cyclic Nucleotide Phosphodiesterases* , 1998, The Journal of Biological Chemistry.

[36]  Development potential of bovine oocytes matured in vitro or in vivo. , 1987 .

[37]  J. Daly,et al.  4-(3-Cyclopentyloxy-4-methoxyphenyl)-2-pyrrolidone (ZK 62711): a potent inhibitor of adenosine cyclic 3',5'-monophosphate phosphodiesterases in homogenates and tissue slices from rat brain. , 1976, Molecular pharmacology.

[38]  M. Sirard,et al.  In vitro inhibition of oocyte nuclear maturation in the bovine. , 1988, Biology of reproduction.

[39]  M. Conti,et al.  Role of Phosphodiesterase Type 3A in Rat Oocyte Maturation1 , 2001, Biology of reproduction.

[40]  R. Stouffer,et al.  The phosphodiesterase 3 inhibitor ORG 9935 inhibits oocyte maturation in the naturally selected dominant follicle in rhesus macaques. , 2008, Contraception.

[41]  S. Palmer,et al.  Pharmacological inhibition of phosphodiesterase 4 triggers ovulation in follicle-stimulating hormone-primed rats. , 2005, Endocrinology.

[42]  R. Gilchrist,et al.  Effect of Specific Phosphodiesterase Isoenzyme Inhibitors During In Vitro Maturation of Bovine Oocytes on Meiotic and Developmental Capacity1 , 2004, Biology of reproduction.

[43]  H. Hidaka,et al.  Selective inhibitor of platelet cyclic adenosine monophosphate phosphodiesterase, cilostamide, inhibits platelet aggregation. , 1979, The Journal of pharmacology and experimental therapeutics.

[44]  K. Fujishige,et al.  Cloning and Characterization of a Novel Human Phosphodiesterase That Hydrolyzes Both cAMP and cGMP (PDE10A)* , 1999, The Journal of Biological Chemistry.

[45]  G. Pincus,et al.  THE COMPARATIVE BEHAVIOR OF MAMMALIAN EGGS IN VIVO AND IN VITRO , 1935, The Journal of experimental medicine.

[46]  J. Cheng,et al.  Isolation and characterization of PDE8A, a novel human cAMP-specific phosphodiesterase. , 1998, Biochemical and biophysical research communications.

[47]  G. Baillie,et al.  RNA Silencing Identifies PDE4D5 as the Functionally Relevant cAMP Phosphodiesterase Interacting with βArrestin to Control the Protein Kinase A/AKAP79-mediated Switching of the β2-Adrenergic Receptor to Activation of ERK in HEK293B2 Cells* , 2005, Journal of Biological Chemistry.

[48]  J. Beavo,et al.  Cyclic Nucleotide Phosphodiesterases: Molecular Regulation to Clinical Use , 2006, Pharmacological Reviews.

[49]  Molecular cloning and characterization of a distinct human phosphodiesterase gene family: PDE11A. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[50]  S. M. Seidel,et al.  Manual of the International Embryo Transfer Society , 1998 .

[51]  K. Saeki,et al.  Developmental capacity of bovine oocytes following inhibition of meiotic resumption by cycloheximide or 6-dimethylaminopurine. , 1997, Theriogenology.

[52]  Chengyu Liu,et al.  Cyclic nucleotide phosphodiesterase 3A-deficient mice as a model of female infertility. , 2004, The Journal of clinical investigation.

[53]  R. Gilchrist,et al.  Differential effects of specific phosphodiesterase isoenzyme inhibitors on bovine oocyte meiotic maturation. , 2002, Developmental biology.

[54]  M. Conti.  Specificity of the Cyclic Adenosine 3′,5′-Monophosphate Signal in Granulosa Cell Function , 2002, Biology of reproduction.