Título:	Bloqueio meiótico como alternativa para aumentar a produção de embriões clone por transferência nuclear
Outros títulos:	Meiotic arrest as an alternative to increase production of cloned embryos by nuclear transfer
Autor(es):	Caixeta, Felippe Manoel Costa
Orientador(es):	Pivato, Ivo
Assunto:	Transferência nuclear Retenção meiótica Clonagem
Data de publicação:	28-Jul-2016
Data de defesa:	5-Fev-2016
Referência:	CAIXETA, Felippe Manoel Costa. Bloqueio meiótico como alternativa para aumentar a produção de embriões clone por transferência nuclear. 2016. xvi, 59 f. Dissertação (Mestrado em Ciências Animais)—Universidade de Brasília, Brasília, 2016.
Abstract:	This study aimed to evaluate the effect of meiotic arrest using phosphodiesterase type 3A (PDE 3A) inhibitors in the production of clones embryos. Two experiments were performed using two inhibitors of meiotic resumption: cilostamide, a specific inhibitor of phosphodiesterase type 3 (PDE3A), and natriuretic peptide type C (NPPC), a peptide that induces the production of guanylate cyclase (cGMP ) that blocks the activity of PDE3A. Oocytes obtained from slaughterhouse ovaries were divided into two groups for each experiment: control (maturation for 20 hours) and Pre-Maturarion (PM) for 6 hours with 5μM of cilostamide or 100 nM of NPPC and then maturation for 20 hours. The oocytes were evaluated for nuclear maturation, embryonic development after in vitro embryo production (IVP),and after SCNT, and total cells number. For SCNT evaluation in each treatment a control group that was activated parthenogenetically (PA) was added. The results were evaluated by Chi-square and Kruskal-Wallis test with P <0.05 considered significant. In the first experiment, where the PM was carried out with 5μM of cilostamide, approximately 87.8% of the oocytes that have undergone PM remained at germinal vesicle stage (GV) after 6 hours of PM, confirming the meiotic block. Embryo development in IVP was similar (P> 0.05) between control and PM, either for cleavage rate on D2 (78.2% and 76.9%) or blastocyst rate on D7 (35, 5% to 29.3%). After SCTN, cleavage rate on D2 was also similar (P>0.05) between control and PM (66% and 51.9%) however, blastocyst rate on D7, was much lower (P<0.05) in PM than on control group (7.4% and 30.2%). In the second experiment, NPPC was used for 6 hours of PM. After PM approximately 84.9% of the oocytes remained at VG stage. Results of IVP showed no difference between control and PM on cleavage (76.4% and 77.8%) and blastocyst rates (48.3% and 45.6%). Similarly, no diferences between PM and control groups were observed for cleavage (69.2% and 68.4%) and blastocyst (24.4% and 21.5%) rates. SCNT and PA embryos from control or PM oocytes showed no diferences in the total cell number. It can be concluded that PM for 6 hours with 100nM NPPC is feasible for embryos cloned production without deleterious effect on the outcome and embryo.
Unidade Acadêmica:	Faculdade de Agronomia e Medicina Veterinária (FAV)
Informações adicionais:	Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, 2016.
Programa de pós-graduação:	Programa de Pós-Graduação em Ciências Animais
Licença:	A concessão da licença deste item refere-se ao termo de autorização impresso assinado pelo autor com as seguintes condições: Na qualidade de titular dos direitos de autor da publicação, autorizo a Universidade de Brasília e o IBICT a disponibilizar por meio dos sites www.bce.unb.br, www.ibict.br, http://hercules.vtls.com/cgi-bin/ndltd/chameleon?lng=pt&skin=ndltd sem ressarcimento dos direitos autorais, de acordo com a Lei nº 9610/98, o texto integral da obra disponibilizada, conforme permissões assinaladas, para fins de leitura, impressão e/ou download, a título de divulgação da produção científica brasileira, a partir desta data.
DOI:	http://dx.doi.org/10.26512/2016.02.D.21047
Aparece nas coleções:	Teses, dissertações e produtos pós-doutorado

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