Participation of lactate dehydrogenase in the in vitro capacitation of fresh and cryopreserved porcine spermatozoa

PEREYRA V; PARIS DUPRAT ML; RODRIGUEZ PC; SATORRE MM; BREININGER E

Congreso; 3ra. Reunión Conjunta de Sociedades de Biología de la República Argentina; 2015

Cryopreserved porcine sperm utilization in biotechnological processes is very limited and there are few studies on the relation betweenenzyme activity and the processes that lead to the acquisition of fertilizing capacity. The aim of this study was to determine theactivity of lactate dehydrogenase (LDH; 1.1.1.27) in fresh and cryopreserved porcine spermatozoa and study its participation in theprocess of in vitro capacitation. LDH activity was determined spectrophotometrically and the enzyme unit (EU) was defined as theamount of LDH enzyme that oxidizes 1 µmol of NADH/minute. Sperm capacitation percentages were determined in the presence orabsence of different concentrations of sodium oxamate (a competitive inhibitor of LDH) by the chlortetracicline technique. Viabilityand motility were also evaluated by the eosin/nigrosin technique and optic microscopy in thermal stage, respectively. The activity ofLDH was 3.40±1.21 EU/1010 spermatozoa in fresh sperm and 0.92±0.67 EU/1010 spermatozoa in cryopreserved sperm. The addition ofthe competitive inhibitor of the enzyme significantly diminished capacitation and motility, at different concentrations in fresh orcryopreserved sperm. Our results demonstrate that lactate dehydrogenase participates in the in vitro capacitation in fresh and cryopreserved spermatozoa and its activity decreases in cryopreservation. The determination of the activity of LDH could be used as amarker of congelability in this species.