title: Nucleoside triphosphate diphosphohydrolase-1 ectonucleotidase is required for normal vas deferens contraction and male fertility through maintaining P2X1 receptor function.
abstract: In this work, we report that Entpd1(-/-) mice, deficient for the ectonucleotidase nucleoside triphosphate diphosphohydrolase-1 (NTPDase1), produce smaller litters (27% reduction) compared with wild-type C57BL6 animals. This deficit is linked to reduced in vivo oocyte fertilization by Entpd1(-/-) males (61 ± 11% versus 88 ± 7% for Entpd1(+/+)). Normal epididymal sperm count, spermatozoa morphology, capacitation, and motility and reduced ejaculated sperm number (2.4 ± 0.5 versus 3.7 ± 0.4 million for Entpd1(+/+)) pointed to vas deferens dysfunction. NTPDase1 was localized by immunofluorescence in the tunica muscularis of the vas deferens. Its absence resulted in a major ATP hydrolysis deficiency, as observed in situ by histochemistry and in primary smooth muscle cell cultures. In vitro, Entpd1(-/-) vas deferens displayed an exacerbated contraction to ATP, a diminished response to its non-hydrolysable analog αβMeATP, and a reduced contraction to electrical field stimulation, suggesting altered P2X1 receptor function with a propensity to desensitize. This functional alteration was accompanied by a 3-fold decrease in P2X1 protein expression in Entpd1(-/-) vas deferens with no variation in mRNA levels. Accordingly, exogenous nucleotidase activity was required to fully preserve P2X1 receptor activation by ATP in vitro. Our study demonstrates that NTPDase1 is required to maintain normal P2X1 receptor functionality in the vas deferens and that its absence leads to impaired peristalsis, reduced spermatozoa concentration in the semen, and, eventually, reduced fertility. This suggests that alteration of NTPDase1 activity affects ejaculation efficacy and male fertility. This work may contribute to unveil a cause of infertility and open new therapeutic potentials.
authors.affiliation: From the Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada, the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada, the Unité mixte de recherche CNRS 6214 INSERM U1083, Université d'Angers, 49045 Angers, France, gilles.kauffenstein@gmail.com.
authors.forename: Gilles
authors.initials: G
authors.lastname: Kauffenstein
authors.affiliation: the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada.
authors.forename: Julie
authors.initials: J
authors.lastname: Pelletier
authors.affiliation: From the Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada, the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada.
authors.forename: Elise G
authors.initials: EG
authors.lastname: Lavoie
authors.affiliation: From the Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada, the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada.
authors.forename: Filip
authors.initials: F
authors.lastname: Kukulski
authors.affiliation: From the Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada, the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada, the Departament de Patologia i Terapèutica Experimental, Facultat de Medicina, Universitat de Barcelona, 08907 Barcelona, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Martín-Satué
authors.affiliation: the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada, the Département de Réadaptation, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada.
authors.forename: Sébastien S
authors.initials: SS
authors.lastname: Dufresne
authors.affiliation: the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada, the Département de Réadaptation, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Frenette
authors.affiliation: From the Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada, the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada.
authors.forename: Cristina
authors.initials: C
authors.lastname: Ribas Fürstenau
authors.affiliation: the Department of Cell Physiology and Pharmacology, University of Leicester, Leicester LE1 9HN, United Kingdom, and.
authors.forename: Michal J
authors.initials: MJ
authors.lastname: Sereda
authors.affiliation: the Unité mixte de recherche CNRS 6214 INSERM U1083, Université d'Angers, 49045 Angers, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Toutain
authors.affiliation: the Unité mixte de recherche CNRS 6214 INSERM U1083, Université d'Angers, 49045 Angers, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Henrion
authors.affiliation: the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada, the Department of Obstetrics, Gynecology, and Reproduction, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada.
authors.forename: Robert
authors.initials: R
authors.lastname: Sullivan
authors.affiliation: the Department of Cell Physiology and Pharmacology, University of Leicester, Leicester LE1 9HN, United Kingdom, and.
authors.forename: Catherine
authors.initials: C
authors.lastname: Vial
authors.affiliation: From the Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec G1V 0A6, Canada, the Centre de Recherche du Centre Hospitalier Universitaire de Québec, Québec G1V 4G2, Canada, Jean.Sevigny@crchul.ulaval.ca.
authors.forename: Jean
authors.initials: J
authors.lastname: Sévigny
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D001081:Apyrase / Q000172:deficiency / Q000235:genetics*; D004542:Ejaculation; D004822:Epididymis / Q000201:enzymology / Q000503:physiopathology; D005260:Female; D005786:Gene Expression Regulation; D007248:Infertility, Male / Q000201:enzymology / Q000235:genetics* / Q000503:physiopathology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009119:Muscle Contraction; D009130:Muscle, Smooth / Q000201:enzymology / Q000503:physiopathology; D009845:Oligospermia / Q000201:enzymology / Q000235:genetics* / Q000503:physiopathology; D009865:Oocytes / Q000502:physiology; D058474:Receptors, Purinergic P2X1 / Q000235:genetics* / Q000378:metabolism; D013075:Sperm Capacitation; D013094:Spermatozoa / Q000502:physiology*; D014649:Vas Deferens / Q000201:enzymology* / Q000503:physiopathology
chemical_list: D015703:Antigens, CD; D058474:Receptors, Purinergic P2X1; D000255:Adenosine Triphosphate; D001081:Apyrase; C067869:CD39 antigen
keywords: ATP; ATPase; Male Fertility; Mouse; NTPDase1/CD39; Nucleotide; P2X1; Reproduction; Smooth Muscle Cells; Vas Deferens