Effects of N-acetylcysteine on Growth, Viability, and Ultrastructure of In Vitro Cultured Bovine Secondary Follicles
Danisvânia R. Nascimento,
Venância A. N. Azevedo,
Pedro A. A. Barroso,
Laryssa G. Barrozo,
Bianca R. Silva,
Anderson W. B. Silva,
Mariana A. M. Donato,
Christina A. Peixoto,
José R. V. Silva
Affiliations
Danisvânia R. Nascimento
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Comandante Maurocélio Rocha Ponte 100, Sobral CEP 62041-040, CE, Brazil
Venância A. N. Azevedo
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Comandante Maurocélio Rocha Ponte 100, Sobral CEP 62041-040, CE, Brazil
Pedro A. A. Barroso
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Comandante Maurocélio Rocha Ponte 100, Sobral CEP 62041-040, CE, Brazil
Laryssa G. Barrozo
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Comandante Maurocélio Rocha Ponte 100, Sobral CEP 62041-040, CE, Brazil
Bianca R. Silva
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Comandante Maurocélio Rocha Ponte 100, Sobral CEP 62041-040, CE, Brazil
Anderson W. B. Silva
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Comandante Maurocélio Rocha Ponte 100, Sobral CEP 62041-040, CE, Brazil
Mariana A. M. Donato
Laboratory of Ultrastructure, CPqAM/FIOCRUZ, Federal University of Pernambuco, Recife CEP 50670-901, PE, Brazil
Christina A. Peixoto
Laboratory of Ultrastructure, CPqAM/FIOCRUZ, Federal University of Pernambuco, Recife CEP 50670-901, PE, Brazil
José R. V. Silva
Laboratory of Biotechnology and Physiology of Reproduction (LABIREP), Federal University of Ceara, Av. Comandante Maurocélio Rocha Ponte 100, Sobral CEP 62041-040, CE, Brazil
This study aimed to investigate the effects of different concentrations of N-acetylcysteine (NAC) on the growth, antrum formation, viability, and ultrastructure of bovine secondary follicles cultured in vitro for 18 days. To this end, the follicles were cultured in TCM-199+ medium alone or supplemented with 1.0, 5.0, or 25.0 mM NAC. Follicular growth, antrum formation, viability (calcein-AM and ethidium homodimer-1) and ultrastructure were evaluated at the end of culture period. The results showed that 1.0 mM NAC increased the percentage of growing follicles and the fluorescence intensity for calcein-AM when compared to other treatments (p < 0.05). On the other hand, follicles cultured with 25.0 mM NAC had higher fluorescence intensity for ethidium homodimer-1, which is a sign of degeneration. Ultrastructural analysis showed that oocytes from follicles cultured in control medium alone or with 1 mM NAC had intact zonae pellucidae in close association with oolemmae, but the ooplasm showed mitochondria with a reduced number of cristae. On the other hand, oocytes from follicles cultured with 5 or 25 mM NAC had extremely vacuolated cytoplasm and no recognizable organelles. In conclusion, 1 mM NAC increases cytoplasmic calcein staining and the growth rate in bovine secondary follicles cultured in vitro, but the presence of 5 or 25 mM NAC causes damage in cellular membranes and organelles, as well as reducing the percentages of growing follicles.
