title: The tyrosine kinase FER is responsible for the capacitation-associated increase in tyrosine phosphorylation in murine sperm.
abstract: Sperm capacitation is required for fertilization. At the molecular level, this process is associated with fast activation of protein kinase A. Downstream of this event, capacitating conditions lead to an increase in tyrosine phosphorylation. The identity of the tyrosine kinase(s) mediating this process has not been conclusively demonstrated. Recent experiments using stallion and human sperm have suggested a role for PYK2 based on the use of small molecule inhibitors directed against this kinase. However, crucially, loss-of-function experiments have not been reported. Here, we used both pharmacological inhibitors and genetically modified mice models to investigate the identity of the tyrosine kinase(s) mediating the increase in tyrosine phosphorylation in mouse sperm. Similar to stallion and human, PF431396 blocks the capacitation-associated increase in tyrosine phosphorylation. Yet, sperm from Pyk2(-/-) mice displayed a normal increase in tyrosine phosphorylation, implying that PYK2 is not responsible for this phosphorylation process. Here, we show that PF431396 can also inhibit FER, a tyrosine kinase known to be present in sperm. Sperm from mice targeted with a kinase-inactivating mutation in Fer failed to undergo capacitation-associated increases in tyrosine phosphorylation. Although these mice are fertile, their sperm displayed a reduced ability to fertilize metaphase II-arrested eggs in vitro.
authors.affiliation: Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA.
authors.forename: Antonio
authors.initials: A
authors.lastname: Alvau
authors.affiliation: Instituto de Biología y Medicina Experimental (IBYME-CONICET), Buenos Aires C1428ADN, Argentina.
authors.forename: Maria Agustina
authors.initials: MA
authors.lastname: Battistone
authors.affiliation: Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA.
authors.forename: Maria Gracia
authors.initials: MG
authors.lastname: Gervasi
authors.affiliation: Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA.
authors.forename: Felipe A
authors.initials: FA
authors.lastname: Navarrete
authors.affiliation: Department of Electrical and Computer Engineering and School of Biomedical Engineering, Colorado State University, Fort Collins, CO 80521, USA.
authors.forename: Xinran
authors.initials: X
authors.lastname: Xu
authors.affiliation: Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca 62210, México.
authors.forename: Claudia
authors.initials: C
authors.lastname: Sánchez-Cárdenas
authors.affiliation: Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca 62210, México.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: De la Vega-Beltran
authors.affiliation: Instituto de Biología y Medicina Experimental (IBYME-CONICET), Buenos Aires C1428ADN, Argentina.
authors.forename: Vanina G
authors.initials: VG
authors.lastname: Da Ros
authors.affiliation: Department of Pathology and Molecular Medicine, Queen's University, Kingston, Ontario, Canada, K7L 3N6.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Greer
authors.affiliation: Departamento de Genética del Desarrollo y Fisiología Molecular, IBT-UNAM, Cuernavaca 62210, México.
authors.forename: Alberto
authors.initials: A
authors.lastname: Darszon
authors.affiliation: Department of Electrical and Computer Engineering and School of Biomedical Engineering, Colorado State University, Fort Collins, CO 80521, USA.
authors.forename: Diego
authors.initials: D
authors.lastname: Krapf
authors.affiliation: Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA.
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Salicioni
authors.affiliation: Instituto de Biología y Medicina Experimental (IBYME-CONICET), Buenos Aires C1428ADN, Argentina.
authors.forename: Patricia S
authors.initials: PS
authors.lastname: Cuasnicu
authors.affiliation: Department of Veterinary and Animal Science, Integrated Sciences Building, University of Massachusetts, Amherst, MA 01003, USA pvisconti@vasci.umass.edu.
authors.forename: Pablo E
authors.identifier: 0000-0001-9320-7518
authors.initials: PE
authors.lastname: Visconti
mesh_terms: D000818:Animals; D051418:Focal Adhesion Kinase 2 / Q000378:metabolism; D008297:Male; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013075:Sperm Capacitation / Q000502:physiology*; D013094:Spermatozoa / Q000201:enzymology*
chemical_list: C052602:proto-oncogene protein c-fes-fps; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases; D051418:Focal Adhesion Kinase 2; C496053:Ptk2b protein, mouse
keywords: Capacitation; FER; Tyrosine phosphorylation