EFFECTS OF SOLUBLE AND MEMBRANE ADENYLATE CYCLASE INHIBITION IN HEPARIN CAPACITATION OF BOVINE SPERMATOZOA

CÓRDOBA MARIANA; FERNÁNDEZ SILVINA

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Jornada; XXIV JORNADAS ANUALES DE LA SOCIEDAD ARGENTINA DE BIOLOGÍA ?Desde la investigación básica a la salud pública: un camino de ida y vuelta?; 2022

Sociedad Argentina de Biologia

EFFECTS OF SOLUBLE AND MEMBRANE ADENYLATE CYCLASE INHIBITION IN HEPARIN CAPACITATION OF BOVINE SPERMATOZOA Fernández S1,3, Córdoba M1,2,31Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal (INITRA). Buenos Aires, Argentina. 2CONICET-Universidad de Buenos Aires. Unidad Ejecutora de Investigaciones en Producción Animal (INPA). Buenos Aires, Argentina. 3Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Química Biológica. Buenos Aires, Argentina. mcordoba@fvet.uba.arIn heparin-induced sperm capacitation, intracellular signals are activated with a specific binding of the inducer with its membrane receptor, involving the participation of different key enzymes such as adenylate cyclase, producing a respiratory burst in the mitochondria. Soluble and membrane isoenzymes of adenylate cyclase could have a different degree of participation in the intracellular signaling mechanism during this process. The aim of this work was to determine the participation of adenylate cyclase isoenzymes in the signal transduction mechanism induced by in vitro heparin capacitation in cryopreserved bovine spermatozoa. Heparin was used as capacitation inducer, LRE-1 as soluble adenylate cyclase inhibitor, and 2,5-dideoxyadenosine (2,5-D) as membrane adenylate cyclase inhibitor. Five treatments were performed with thawed semen in TALP medium at 38°C: control, heparin, heparin/LRE-1, heparin/2,5-D, and heparin/LRE-1/2,5-D. Capacitation was evaluated by the chlorotetracycline epifluorescent technique and membrane viability and integrity by trypan blue vital staining with differential interferential contrast. Sperm motility was evaluated by microscopy and analyzed with the ISAS-Prosier software. Mitochondrial membrane potential was measured using the fluorochrome JC-1. Data were analyzed by ANOVA and Tukey´s test (P