Heparin- and superoxide anion-dependent capacitation of cryopreserved bovine spermatozoa: requirement of dehydrogenases and protein kinases.
Capacitation is part of an oxidative process necessary for bovine spermatozoa to acquire fertilizing capacity. This process includes the generation of reactive oxygen species (ROS) and the participation of protein kinases such as A (PKA), C (PKC) and tyrosine kinase (PTK). A redox status is required to support both sperm motility and capacitation. Our aim was to determine the requirement of lactate dehydrogenase C4 (LDH-C4) and isocitrate dehydrogenase (NADP-ICDH) and of protein kinases in cryopreserved bovine sperm capacitation. The presence of inhibitors of both LDH-C4 and NADP-ICDH prevented the heparin-induced capacitation. H89, GF109203X or genistein blocked capacitation triggered by heparin or the superoxide (O(-*)(2))generator system xanthine-xanthine oxidase-catalase (XXOC) suggesting the requirement of PKA, PKC and PTK in this process. Taken together these results suggest that LDH-C4 and NADP-ICDH contribute with the redox status to support bovine sperm capacitation and that PKA, PKC and PTK are involved in different mechanisms induced by different inducers that lead bovine spermatozoa to be capacitated.
O'Flaherty
Cristian
C
Area of Biochemistry, School of Veterinary Sciences, University of Buenos Aires, Buenos Aires, Argentina. cristian.oflaherty@mcgill.ca
Beorlegui
Norma
N
Beconi
Martha T
MT
D000818:Animals; D002417:Cattle; D015925:Cryopreservation / Q000662:veterinary; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005343:Fibrinolytic Agents / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D008297:Male; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D012662:Semen Preservation; D013075:Sperm Capacitation / Q000187:drug effects / Q000502:physiology*; D013481:Superoxides / Q000494:pharmacology
D004791:Enzyme Inhibitors; D005343:Fibrinolytic Agents; D013481:Superoxides; D006493:Heparin; D010088:Oxidoreductases; D011494:Protein Kinases