title: Active 3'-5' cyclic nucleotide phosphodiesterases are present in detergent-resistant membranes of mural granulosa cells.
abstract: Lipids rafts are specialised membrane microdomains involved in cell signalling that can be isolated as detergent-resistant membranes (DRMs). The second messenger cyclic AMP (cAMP) has a central role in cell signalling in the ovary and its degradation is carried out by the phosphodiesterase (PDE) enzyme family. We hypothesised that PDEs could be functionally present in the lipid rafts of porcine mural granulosa cell membranes. PDE6C, PDE8A and PDE11A were detected by dot blot in the DRMs and the Triton-soluble fraction of the mural granulosa cells membrane and the cytosol. As shown by immunocytochemistry, PDEs showed clear immunostaining in mural granulosa cell membranes and the cytosol. Interestingly, cAMP-PDE activity was 18 times higher in the DRMs than in the Triton-soluble fraction of cell membranes and was 7.7 times higher in the cytosol than in the DRMs. cAMP-PDE activity in mural granulosa cells was mainly contributed by the PDE8 and PDE11 families. This study shows that PDEs from the PDE8 and PDE11 families are present in mural granulosa cells and that the cAMP-PDE activity is mainly contributed by the cytosol. In the cell membrane, the cAMP-PDE activity is mainly contributed by the DRMs. In addition, receptors for prostaglandin E2 and LH, two G-protein-coupled receptors, are present in lipid rafts and absent from the non-raft fraction of the granulosa cell membrane. These results suggest that in these cells, the lipid rafts exist as a cell-signalling platform and PDEs are one of the key enzyme families present in the raft.
authors.affiliation: Centre de Recherche en Biologie de la Reproduction, Département des Sciences Animales, Faculté des Sciences de l'Agriculture et de l'Alimentation, 2425 rue de l'Agriculture, Pavillon Paul-Comtois, Université Laval, Québec, G1V 0A6, Canada.
authors.forename: Annick
authors.initials: A
authors.lastname: Bergeron
authors.affiliation: Centre de Recherche en Biologie de la Reproduction, Département des Sciences Animales, Faculté des Sciences de l'Agriculture et de l'Alimentation, 2425 rue de l'Agriculture, Pavillon Paul-Comtois, Université Laval, Québec, G1V 0A6, Canada.
authors.forename: Christine
authors.initials: C
authors.lastname: Guillemette
authors.affiliation: Centre de Recherche en Biologie de la Reproduction, Département des Sciences Animales, Faculté des Sciences de l'Agriculture et de l'Alimentation, 2425 rue de l'Agriculture, Pavillon Paul-Comtois, Université Laval, Québec, G1V 0A6, Canada.
authors.forename: Marc-André
authors.initials: MA
authors.lastname: Sirard
authors.affiliation: Centre de Recherche en Biologie de la Reproduction, Département des Sciences Animales, Faculté des Sciences de l'Agriculture et de l'Alimentation, 2425 rue de l'Agriculture, Pavillon Paul-Comtois, Université Laval, Québec, G1V 0A6, Canada.
authors.forename: François J
authors.initials: FJ
authors.lastname: Richard
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000201:enzymology*; D002784:Cholesterol / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D005260:Female; D006107:Granulosa Cells / Q000201:enzymology*; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D013552:Swine
chemical_list: D002784:Cholesterol; D000242:Cyclic AMP; D010727:Phosphoric Diester Hydrolases