Role of two glycosidases (alpha-mannosidase and beta-N-acetylglucosaminidase) on in vitro bovine embryonic development.

Glycosidases are enzymes that might play a role in embryonic development. The aims of the present project were to evaluate if bovine in vitro produced embryos: (1) release beta-N-acetylglucosaminidase (beta-NAGASE) and alpha-mannosidase in culture medium and (2) to investigate if these glycosidases may be used as markers of embryo quality. Bovine embryos were obtained using routine methods for IVM, IVF and IVC. Two experiments were done [(experiment 1: culture of embryos in the same droplet until day 7 and experiment 2: separation and transfer of embryos to new droplets at the morula stage (day 6)]. Samples were collected on day 7 (experiment 1) and on days 6 and 7 (experiment 2). The results of the present study are summarized as follow: (i) Embryos release both glycosidases. (ii) The activity of both glycosidases was significantly lower (p<0.05) in droplets with degenerate embryos compared to droplets without degenerate embryos. (iii) The activity of beta-NAGASE was higher in droplets which contained morulae compared to droplets without morulae. In conclusion, embryos release both glucosidases during their development, while degenerate embryos release less beta-NAGASE and alpha-mannosidase compared to good embryos. Furthermore, beta-NAGASE secretion seems to be related to retarded morulae.