title: Participation of phosphofructokinase, malate dehydrogenase and isocitrate dehydrogenase in capacitation and acrosome reaction of boar spermatozoa.
abstract: The aim of this work was to determine the enzymatic activity of phosphofructokinase (PFK), malate dehydrogenase (MDH) and isocitrate dehydrogenase (IDH) in boar spermatozoa and study their participation in bicarbonate-induced capacitation and follicular fluid-induced acrosome reaction. Enzymatic activity of these enzymes was determined spectrophotometrically in extracts of boar spermatozoa. Sperm suspensions were incubated in the presence of bicarbonate (40 mM), a well-known capacitation inducer, or follicular fluid (30%), as an acrosome reaction inducer, and different concentrations of oxoglutarate, oxalomalate and hydroxymalonate, inhibitors of PFK, IDH and MDH, respectively. Capacitation percentages were determined by the fluorescence technique of chlortetracycline (CTC), and true acrosome reaction was determined by trypan blue and differential-interferential contrast, optical microscopy. The activity of PFK in boar spermatozoa enzymatic extracts was 1.70 ± 0.19 U/1010spermatozoa, the activity of NAD- and NADP-dependent IDH was 0.111 ± 0.005 U/1010and 2.22 ± 0.14 U/1010spermatozoa, respectively, and the activity of MDH was 4.24 ± 0.38 U/1010spermatozoa. The addition of the specific inhibitors of these enzymes prevented sperm capacitation and decreased sperm motility during capacitation and inhibited the acrosome reaction (AR), without affecting the sperm motility during this process. Our results demonstrate the participation of PFK, IDH and MDH in bicarbonate-induced capacitation and follicular fluid-induced acrosome reaction in boar spermatozoa, contributing to elucidate the mechanisms that produce energy necessary for these processes in porcine spermatozoa.
authors.affiliation: Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Universidad de Buenos Aires, Buenos Aires, Argentina.|Investigaciones en Producción Animal (INPA), CONICET - Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: E
authors.identifier: http://orcid.org/0000-0001-7185-0190
authors.initials: E
authors.lastname: Breininger
authors.affiliation: Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: D
authors.initials: D
authors.lastname: Dubois
authors.affiliation: Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: V E
authors.initials: VE
authors.lastname: Pereyra
authors.affiliation: Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: P C
authors.initials: PC
authors.lastname: Rodriguez
authors.affiliation: Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: M M
authors.initials: MM
authors.lastname: Satorre
authors.affiliation: Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Universidad de Buenos Aires, Buenos Aires, Argentina.|Investigaciones en Producción Animal (INPA), CONICET - Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: P D
authors.initials: PD
authors.lastname: Cetica
mesh_terms: D020101:Acrosome Reaction / Q000187:drug effects*; D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology; D005260:Female; D015571:Follicular Fluid / Q000502:physiology; D007521:Isocitrate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008297:Male; D025221:Phosphofructokinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013075:Sperm Capacitation / Q000187:drug effects*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology*; D034421:Sus scrofa; D013646:Tartronates / Q000494:pharmacology
chemical_list: D001639:Bicarbonates; D013646:Tartronates; D008291:Malate Dehydrogenase; D007521:Isocitrate Dehydrogenase; D025221:Phosphofructokinases