title: Identification and Localization of the Cyclic Nucleotide Phosphodiesterase 10A in Bovine Testis and Mature Spermatozoa.
abstract: In mammals, adenosine 3', 5'-cyclic monophosphate (cAMP) is known to play highly important roles in sperm motility and acrosomal exocytosis. It is known to act through protein phosphorylation via PRKA and through the activation of guanine nucleotide exchange factors like EPAC. Sperm intracellular cAMP levels depend on the activity of adenylyl cyclases, mostly SACY, though transmembrane-containing adenylyl cyclases are also present, and on the activity of cyclic nucleotide phosphodiesterases (PDE) whose role is to degrade cAMP into 5'-AMP. The PDE superfamily is subdivided into 11 families (PDE1 to 11), which act on either cAMP or cGMP, or on both cAMP and cGMP although with different enzymatic properties. PDE10, which is more effective on cAMP than cGMP, has been known for almost 15 years and is mostly studied in the brain where it is associated with neurological disorders. Although a high level of PDE10A gene expression is observed in the testis, information on the identity of the isoforms or on the cell type that express the PDE10 protein is lacking. The objective of this study was to identify the PDE10A isoforms expressed in the testis and germ cells, and to determine the presence and localization of PDE10A in mature spermatozoa. As a sub-objective, since PDE10A transcript variants were reported strictly through analyses of bovine genomic sequence, we also wanted to determine the nucleotide and amino acid sequences by experimental evidence. Using RT-PCR, 5'- and 3'-RACE approaches we clearly show that PDE10A transcript variants X3 and X5 are expressed in bovine testis as well as in primary spermatocytes and spermatids. We also reveal using a combination of immunological techniques and proteomics analytical tools that the PDE10A isoform X4 is present in the area of the developing acrosome of spermatids and of the acrosome of mature spermatozoa.
authors.affiliation: Département d'obstétrique, gynécologie et reproduction, Université Laval, et Centre de recherche du CHU de Québec-Université Laval, Québec, Canada.|Centre de recherche en reproduction, développement et santé intergénérationnelle (CRDSI), Université Laval, Québec, Canada.|Centre de recherche du CHU de Québec-Université Laval, G1V 4G2, Québec, QC, Canada.
authors.forename: Serge
authors.initials: S
authors.lastname: Goupil
authors.affiliation: Département d'obstétrique, gynécologie et reproduction, Université Laval, et Centre de recherche du CHU de Québec-Université Laval, Québec, Canada.|Centre de recherche en reproduction, développement et santé intergénérationnelle (CRDSI), Université Laval, Québec, Canada.|Centre de recherche du CHU de Québec-Université Laval, G1V 4G2, Québec, QC, Canada.
authors.forename: Loïze
authors.initials: L
authors.lastname: Maréchal
authors.affiliation: Département de médecine moléculaire, Université Laval, et Centre de recherche du CHU de Québec-Université Laval, Québec, Canada.|Centre de recherche du CHU de Québec-Université Laval, G1V 4G2, Québec, QC, Canada.
authors.forename: Hassan
authors.initials: H
authors.lastname: El Hajj
authors.affiliation: Département de médecine moléculaire, Université Laval, et Centre de recherche du CHU de Québec-Université Laval, Québec, Canada.|Centre de recherche du CHU de Québec-Université Laval, G1V 4G2, Québec, QC, Canada.
authors.forename: Marie-Ève
authors.initials: MÈ
authors.lastname: Tremblay
authors.affiliation: Département des sciences animales, Université Laval, et Centre de recherche du CHU de Québec-Université Laval, Québec, Canada.|Centre de recherche en reproduction, développement et santé intergénérationnelle (CRDSI), Université Laval, Québec, Canada.
authors.forename: François J
authors.initials: FJ
authors.lastname: Richard
authors.affiliation: Département d'obstétrique, gynécologie et reproduction, Université Laval, et Centre de recherche du CHU de Québec-Université Laval, Québec, Canada.|Centre de recherche en reproduction, développement et santé intergénérationnelle (CRDSI), Université Laval, Québec, Canada.|Centre de recherche du CHU de Québec-Université Laval, G1V 4G2, Québec, QC, Canada.
authors.forename: Pierre
authors.identifier: http://orcid.org/0000-0003-2053-3958
authors.initials: P
authors.lastname: Leclerc
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000235:genetics* / Q000378:metabolism; D020101:Acrosome Reaction / Q000235:genetics; D000262:Adenylyl Cyclases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D000242:Cyclic AMP / Q000378:metabolism*; D006152:Cyclic GMP / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism; D008297:Male; D010766:Phosphorylation; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013079:Sperm Maturation / Q000235:genetics; D013081:Sperm Motility / Q000235:genetics; D013087:Spermatids / Q000201:enzymology* / Q000254:growth & development; D013090:Spermatocytes / Q000201:enzymology* / Q000254:growth & development; D013379:Substrate Specificity; D013737:Testis / Q000166:cytology / Q000201:enzymology / Q000254:growth & development
chemical_list: D020662:Guanine Nucleotide Exchange Factors; D000242:Cyclic AMP; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D000262:Adenylyl Cyclases; D006152:Cyclic GMP