title: The prion-like protein Doppel enhances ovine spermatozoa fertilizing ability.
abstract: The function of prion-like protein Doppel was suggested to be related to male fertility. In this study, the importance of ovine Doppel polypeptide on spermatozoa capacitation and fertilization was evaluated. After refolding, recombinant Doppel (rDpl) was supplemented with different concentrations (40, 80 or 190 ng/ml) to ovine spermatozoa during the capacitation process. In experiment 1, post-thawed ovine spermatozoa were incubated with different concentrations of rDpl during 1 h for swim-up, and changes in sperm motility, concentration, vigour, viability and capacitation were monitored (10 replicates). In experiment 2, the fertilization ability of post-swim-up spermatozoa incubated as above was tested through heterologous fertilization of bovine in vitro matured oocytes (n = 423, three replicates). Regardless of dosage, rDpl improved (p ≤ 0.03) spermatozoa viability. Sperm individual motility and vigour were the highest (p ≤ 0.04) for the group receiving 190 ng/ml rDpl. Sperm supplemented with the highest doses of rDpl achieved higher (p = 0.02) fertilization rates (56.0 ± 3.0%) than control (39.1 ± 2.2%) and 40 ng/ml rDpl (39.8 ± 2.7%). Preliminary data suggest that Doppel protein may enhance in vitro spermatozoa fertilizing ability.
authors.affiliation: Unidade de Recursos Genéticos, Reprodução e Melhoramento Animal, Instituto Nacional dos Recursos Biológicos, Vale de Santarém, Portugal.
authors.forename: J
authors.initials: J
authors.lastname: Pimenta
authors.forename: F M V
authors.initials: FM
authors.lastname: Dias
authors.forename: C C
authors.initials: CC
authors.lastname: Marques
authors.forename: M C
authors.initials: MC
authors.lastname: Baptista
authors.forename: M I
authors.initials: MI
authors.lastname: Vasques
authors.forename: A E M
authors.initials: AE
authors.lastname: Horta
authors.forename: J P
authors.initials: JP
authors.lastname: Barbas
authors.forename: R
authors.initials: R
authors.lastname: Soares
authors.forename: P
authors.initials: P
authors.lastname: Mesquita
authors.forename: E
authors.initials: E
authors.lastname: Cabrita
authors.forename: C M G A
authors.initials: CM
authors.lastname: Fontes
authors.forename: J A
authors.initials: JA
authors.lastname: Prates
authors.forename: R M
authors.initials: RM
authors.lastname: Pereira
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D005307:Fertilization in Vitro / Q000662:veterinary; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D008297:Male; D011328:Prions / Q000235:genetics / Q000378:metabolism*; D034561:Sheep, Domestic / Q000502:physiology*; D013075:Sperm Capacitation / Q000502:physiology; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*
chemical_list: D058851:GPI-Linked Proteins; D011328:Prions