title: Male fertility and apoptosis in normal spermatogenesis are regulated by vacuolar-ATPase isoform a2.
abstract: The a2 isoform of vacuolar-ATPase (ATP6V0A2, referred to as a2V) is required for normal spermatogenesis and maturation of sperm. Treatment of male mice with anti-a2V disturbs the testicular cytokine/chemokine balance and leads to severe deficiencies of spermatogenesis. The aim of the present study was to investigate the role of a2V in male fertility and in the regulation of apoptotic pathways required for normal spermatogenesis in mice. To study the role of a2V single dose of anti-a2V monoclonal antibody or mouse IgG isotype (3μg/animal) was injected i.p. into males on alternate days for 10 days. The expression of sperm maturation-related molecules and pro-apoptotic molecules was measured by real-time PCR or immunohistochemistry in control and anti-a2V-treated testes. The caspase levels and their activity were measured by western blot and fluorometry. We found that the expression of the sperm maturation-related molecules SPAM1, ADAM1, and ADAM2 was significantly decreased in testes from anti-a2V-treated males. The expression of pro-apoptotic molecules (Bax, p53, and p21) and molecules involved in the intrinsic pathway of apoptosis (caspase-9, caspase-3, and PARP), which are crucial for normal spermatogenesis was significantly reduced in testes from anti-a2V-treated males compared with the control. The total ATP level was significantly lower in anti-a2V-treated testes. The data provide novel evidence showing that a2V can regulate the apoptotic pathways, an essential testicular feature, and is necessary for efficient spermatogenesis.
authors.affiliation: Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA. Electronic address: mukesh.jaiswal@rosalindfranklin.edu.
authors.forename: Mukesh K
authors.initials: MK
authors.lastname: Jaiswal
authors.affiliation: Department of Obstetrics and Gynecology, NorthShore University Health System, Evanston, IL, USA.
authors.forename: Varkha
authors.initials: V
authors.lastname: Agrawal
authors.affiliation: Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
authors.forename: Gajendra K
authors.initials: GK
authors.lastname: Katara
authors.affiliation: Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
authors.forename: Sahithi
authors.initials: S
authors.lastname: Pamarthy
authors.affiliation: Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
authors.forename: Arpita
authors.initials: A
authors.lastname: Kulshrestha
authors.affiliation: U976 INSERM /UMR 976CNRS Saint Louis Hospital, 75010 Paris, France.
authors.forename: Gerard
authors.initials: G
authors.lastname: Chaouat
authors.affiliation: Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
authors.forename: Alice
authors.initials: A
authors.lastname: Gilman-Sachs
authors.affiliation: Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
authors.forename: Kenneth D
authors.initials: KD
authors.lastname: Beaman
mesh_terms: D051722:ADAM Proteins / Q000276:immunology; D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins / Q000276:immunology; D020169:Caspases / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000276:immunology; D000072196:Fertilins; D005298:Fertility / Q000187:drug effects / Q000276:immunology*; D006821:Hyaluronoglucosaminidase / Q000276:immunology; D008297:Male; D008562:Membrane Glycoproteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000276:immunology*; D013091:Spermatogenesis / Q000187:drug effects / Q000276:immunology*; D013094:Spermatozoa / Q000276:immunology*
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D051017:Apoptosis Regulatory Proteins; C432920:Atp6v0a2 protein, mouse; D015815:Cell Adhesion Molecules; D008562:Membrane Glycoproteins; D006821:Hyaluronoglucosaminidase; C066852:hyaluronidase PH-20; D020169:Caspases; D051722:ADAM Proteins; C000606458:Adam2 protein, mouse; D000072196:Fertilins; D006180:Proton-Translocating ATPases
keywords: Bax; Bcl2; Caspase-9; Male fertility; Spermatogenesis; Testes; p53