Hyaluronic acid capacitation induces intracellular signals modulated by membrane-associated adenylate cyclase and tyrosine kinase involved in bovine in vitro fertilization

FERNÁNDEZ, S.; MORADO, S.; CETICA, P.; CÓRDOBA, M.

THERIOGENOLOGY

ELSEVIER SCIENCE INC

Año: 2020

0093-691X

Mammalian sperm go through a series of events in the female reproductive tract, known as sperm capacitation, to be capable of fertilizing the oocyte. Although heparin is the most commonly used in vitro capacitation inducer in the bovine, hyaluronic acid (HA) has been recently used for capacitation induction as well as for other reproductive biotechnologies such as sperm selection and in vitro fertilization (IVF).Our aim was to induce sperm capacitation with heparin or HA in order to study membrane-associated adenylate cyclase (mAC) and tyrosine kinase (TK) intracellular signal activation and its relation with the cleavage rate and the oxidative status in potential bovine embryos. Sperm capacitation was induced with HA or heparin, while 2,5-dideoxyadenosine and genistein were used as mAC and TK inhibitors, respectively. Sperm capacitation was studied using CTC technique, sperm plasma membrane and acrosome integritywere determined using trypan blue stain anddifferential interference contrast, and mitochondrial activity was evaluated usingfluorochromeJC-1. Sperm mediated oocyte activation was carried out in IVF-mSOF with frozen-thawed semen. Then, activated oocytes were cultured in IVC medium for 8 days. Cleavage rate was analysed at 48 hours and blastocyst production 7-8 days after IVF. Cytosolic oxidative activity was determined in activated oocytes 7hours after IVF using RedoxSensor Red CC-1 fluorochrome.