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 | Acesso ao texto completo restrito à biblioteca da Embrapa Pecuária Sudeste. Para informações adicionais entre em contato com cppse.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Pecuária Sudeste. |
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Data corrente: |
28/09/2010 |
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Data da última atualização: |
30/06/2023 |
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Tipo da produção científica: |
Resumo em Anais de Congresso |
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Autoria: |
SARAIVA, N. Z.; OLIVEIRA, C. S.; TETZNER, T. A. D.; SOUZA, M. M.; LIMA, M. R. de; NICIURA, S. C. M.; GARCIA, J. M. |
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Afiliação: |
N. Z. SARAIVA, UNESP/JABOTICABAL; C. S. OLIVEIRA, UNESP/JABOTICABAL; T. A. D. TETZNER, UNESP/JABOTICABAL; M. M. SOUZA, UNESP/JABOTICABAL; M. R. DE LIMA, UNESP/JABOTICABAL; SIMONE CRISTINA MEO NICIURA, CPPSE; J. M. GARCIA, UNESP/JABOTICABAL. |
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Título: |
Bovine cytoplasts prepared by demecolcine-induced enucleation of activated oocytes. |
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Ano de publicação: |
2010 |
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Fonte/Imprenta: |
In: ANNUAL CONFERENCE OT THE INTERNATIONAL EMBRYO TRANSFER SOCIETY, 36., 2010, Cordoba. Proceedings... Córdoba: ITES, 2010; Reproduction, Fertility and development, v. 22, n. 1, 2010. |
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Páginas: |
p. 197 |
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Idioma: |
Português |
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Conteúdo: |
One of the most critical steps of the standard NT procedure is the removal of oocyte chromosomes for the production of enucleated cytoplasts. This process involves ultraviolet (UV) irradiation, which causes damage to the membrane and intracellular components of bovine oocytes. The objective of the present study was to evaluate the use of demecolcine, a microtubule depolymerizing agent, for chemical-induced enucleation of activated bovine oocytes. In the first experiment, oocytes obtained from cow ovaries collected in a slaughterhouse were IVM in TCM-199 medium supplemented with 10% FCS, FSH, hCG, estradiol, pyruvate, and amikacin for 26 h; then, they were artificially activated with 5 ?M ionomycin for 5 min and 10 ?g mL-1 cycloheximide for 4 h. The following groups were established: control, C (no activation and no demecolcine); activated, A (exposure only to activating agents); and demecolcine-exposed oocytes, DEME (activation for 4 h and 0.05 ?g mL-1 demecolcine for 2 to 4 h after 0, 0.5, 1.0, 1.5, and 2.0 h of activation). The treatment DEME comprised the groups G1 (demecolcine from 0 to 2 h of activation; DEME 0-2 h); G2 (DEME 0-4 h); G3 (DEME 0.5-2.5 h); G4 (DEME 0.5-4 h); G5 (DEME 1-3 h); G6 (DEME 1-4 h); G7 (DEME 1.5-3.5 h); G8 (DEME 1.5-4 h); and G9 (DEME 2-4h). The oocytes were stained with 10 ?g mL-1 Hoechst 33342 for 15 min and enucleation rates (EN) were evaluated under epifluorescence microscope (330-385 nm).After EN evaluation, a second experiment was performed to verify the efficiency of demecolcine-induced enucleation in group G9, which showed the best result in the first experiment. After demecolcine treatment, the oocytes were incubated in HEPES-buffered SOF with 10% FCS and cytochalasin B for removal of the 2PB and minimal adjacent cytoplast under an inverted optical microscope. Traditional enucleation was performed on the control group without exposure of the oocytes to demecolcine. The same conditions were employed except that UV light was used to confirm enucleation. Samples of cytoplasts were stained with Hoechst for 15 min and analyzed for enucleation efficiency. Five and 3 replicates were evaluated, respectively, in experiments 1 and 2, and the results were analyzed by chi-square test in the statistical software Minitab®, release 14.1 (Minitab Inc., State College, PA, USA). A level of 5% significance was used. Regarding enucleation rates, all treated groups were significantly different compared with the C (0/114) and A (0/130) groups. Considering the DEME groups, G8 (46/109; 42.2%) and G9 (61/113; 54.0%) presented superior rates (P < 0.05) to all other groups (26.8 to 36.3%), but they were similar despite the great tendency (P = 0.07) to difference. We observed high efficiency (P < 0.05) of demecolcine-induced enucleation (90/110; 81.8%) compared with a traditional technique (61/95; 64.2%). In conclusion, the present study shows that demecolcine-induced enucleation of activated oocytes enables good rates of enucleation and has greater efficiency than the traditional technique as well as avoiding UV irradiation of the cytoplast. MenosOne of the most critical steps of the standard NT procedure is the removal of oocyte chromosomes for the production of enucleated cytoplasts. This process involves ultraviolet (UV) irradiation, which causes damage to the membrane and intracellular components of bovine oocytes. The objective of the present study was to evaluate the use of demecolcine, a microtubule depolymerizing agent, for chemical-induced enucleation of activated bovine oocytes. In the first experiment, oocytes obtained from cow ovaries collected in a slaughterhouse were IVM in TCM-199 medium supplemented with 10% FCS, FSH, hCG, estradiol, pyruvate, and amikacin for 26 h; then, they were artificially activated with 5 ?M ionomycin for 5 min and 10 ?g mL-1 cycloheximide for 4 h. The following groups were established: control, C (no activation and no demecolcine); activated, A (exposure only to activating agents); and demecolcine-exposed oocytes, DEME (activation for 4 h and 0.05 ?g mL-1 demecolcine for 2 to 4 h after 0, 0.5, 1.0, 1.5, and 2.0 h of activation). The treatment DEME comprised the groups G1 (demecolcine from 0 to 2 h of activation; DEME 0-2 h); G2 (DEME 0-4 h); G3 (DEME 0.5-2.5 h); G4 (DEME 0.5-4 h); G5 (DEME 1-3 h); G6 (DEME 1-4 h); G7 (DEME 1.5-3.5 h); G8 (DEME 1.5-4 h); and G9 (DEME 2-4h). The oocytes were stained with 10 ?g mL-1 Hoechst 33342 for 15 min and enucleation rates (EN) were evaluated under epifluorescence microscope (330-385 nm).After EN evaluation, a second experiment was performed... Mostrar Tudo |
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Palavras-Chave: |
Bovine; Cytoplasts. |
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Thesaurus Nal: |
oocytes. |
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Categoria do assunto: |
W Química e Física |
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Marc: |
LEADER 03900nam a2200229 a 4500
001 1863081
005 2023-06-30
008 2010 bl uuuu u00u1 u #d
100 1 $aSARAIVA, N. Z.
245 $aBovine cytoplasts prepared by demecolcine-induced enucleation of activated oocytes.$h[electronic resource]
260 $aIn: ANNUAL CONFERENCE OT THE INTERNATIONAL EMBRYO TRANSFER SOCIETY, 36., 2010, Cordoba. Proceedings... Córdoba: ITES, 2010; Reproduction, Fertility and development, v. 22, n. 1, 2010.$c2010
300 $ap. 197
520 $aOne of the most critical steps of the standard NT procedure is the removal of oocyte chromosomes for the production of enucleated cytoplasts. This process involves ultraviolet (UV) irradiation, which causes damage to the membrane and intracellular components of bovine oocytes. The objective of the present study was to evaluate the use of demecolcine, a microtubule depolymerizing agent, for chemical-induced enucleation of activated bovine oocytes. In the first experiment, oocytes obtained from cow ovaries collected in a slaughterhouse were IVM in TCM-199 medium supplemented with 10% FCS, FSH, hCG, estradiol, pyruvate, and amikacin for 26 h; then, they were artificially activated with 5 ?M ionomycin for 5 min and 10 ?g mL-1 cycloheximide for 4 h. The following groups were established: control, C (no activation and no demecolcine); activated, A (exposure only to activating agents); and demecolcine-exposed oocytes, DEME (activation for 4 h and 0.05 ?g mL-1 demecolcine for 2 to 4 h after 0, 0.5, 1.0, 1.5, and 2.0 h of activation). The treatment DEME comprised the groups G1 (demecolcine from 0 to 2 h of activation; DEME 0-2 h); G2 (DEME 0-4 h); G3 (DEME 0.5-2.5 h); G4 (DEME 0.5-4 h); G5 (DEME 1-3 h); G6 (DEME 1-4 h); G7 (DEME 1.5-3.5 h); G8 (DEME 1.5-4 h); and G9 (DEME 2-4h). The oocytes were stained with 10 ?g mL-1 Hoechst 33342 for 15 min and enucleation rates (EN) were evaluated under epifluorescence microscope (330-385 nm).After EN evaluation, a second experiment was performed to verify the efficiency of demecolcine-induced enucleation in group G9, which showed the best result in the first experiment. After demecolcine treatment, the oocytes were incubated in HEPES-buffered SOF with 10% FCS and cytochalasin B for removal of the 2PB and minimal adjacent cytoplast under an inverted optical microscope. Traditional enucleation was performed on the control group without exposure of the oocytes to demecolcine. The same conditions were employed except that UV light was used to confirm enucleation. Samples of cytoplasts were stained with Hoechst for 15 min and analyzed for enucleation efficiency. Five and 3 replicates were evaluated, respectively, in experiments 1 and 2, and the results were analyzed by chi-square test in the statistical software Minitab®, release 14.1 (Minitab Inc., State College, PA, USA). A level of 5% significance was used. Regarding enucleation rates, all treated groups were significantly different compared with the C (0/114) and A (0/130) groups. Considering the DEME groups, G8 (46/109; 42.2%) and G9 (61/113; 54.0%) presented superior rates (P < 0.05) to all other groups (26.8 to 36.3%), but they were similar despite the great tendency (P = 0.07) to difference. We observed high efficiency (P < 0.05) of demecolcine-induced enucleation (90/110; 81.8%) compared with a traditional technique (61/95; 64.2%). In conclusion, the present study shows that demecolcine-induced enucleation of activated oocytes enables good rates of enucleation and has greater efficiency than the traditional technique as well as avoiding UV irradiation of the cytoplast.
650 $aoocytes
653 $aBovine
653 $aCytoplasts
700 1 $aOLIVEIRA, C. S.
700 1 $aTETZNER, T. A. D.
700 1 $aSOUZA, M. M.
700 1 $aLIMA, M. R. de
700 1 $aNICIURA, S. C. M.
700 1 $aGARCIA, J. M.
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Embrapa Pecuária Sudeste (CPPSE) |
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| Registros recuperados : 24 | |
| 6. |  | SARAIVA, N. Z.; PERECIN, F.; MÉO, S. C.; FERREIRA, C. R.; TETZNER, T. A. D.; GARCIA, J. M. Demecolcine effects on microtubule kinetics and chemically assisted enucleation of bovine oocytes. Cloning and Stem Cells, v. 11, n. 1, p. 141-152, mar. 2009.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
|    |
| 7. | | SARAIVA, N. Z.; PERECIN, F.; MÉO, S. C.; FERREIRA, C. R.; TETZNER, T. A. D.; VANTINI, R.; GARCIA, J. M. Efeitos da demecolcina sobre a cinética da maturação nuclear e a migração dos grânulos corticais em oócitos bovinos. Acta Scientiae Veterinariae, v. 35, supl. 3, p. s1276, 2007; In: REUNIÃO ANUAL DA SOCIEDADE BRASILEIRA DE TECNOLOGIA DE EMBRIÕES, 21., 2007, Salvador. Anais... Salvador: UFRGS, 2007. p. 1276.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
|  |
| 8. | | SARAIVA, N. Z.; OLIVEIRA, C. S.; TETZNER, T. A. D.; SOUZA, M. M.; LIMA, M. R. de; NICIURA, S. C. M.; GARCIA, J. M. Bovine cytoplasts prepared by demecolcine-induced enucleation of activated oocytes. In: ANNUAL CONFERENCE OT THE INTERNATIONAL EMBRYO TRANSFER SOCIETY, 36., 2010, Cordoba. Proceedings... Córdoba: ITES, 2010; Reproduction, Fertility and development, v. 22, n. 1, 2010. p. 197| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
| |
| 9. | | SARAIVA, N. Z.; OLIVEIRA, C. S.; TETZNER, T. A. D.; LIMA, M. R. de; MELO, D. S. de; NICIURA, S. C. M.; GARCIA, J. M. Chemically assisted enucleation results in higher G6PD expression in early bovine female embryos obtained by somatic cell nuclear transfer. Cellular Reprogramming, v. 14, n. 5, p. 1-11, 2012.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 2 |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 10. | | TETZNER, T. A. D.; SARAIVA, N. Z.; OLIVEIRA, C. S.; NICIURA, S. C. M.; SOUZA M. M.; LIMA, M. R.; GARCIA J. M. Effects of culture with ovalbumin in absence of fetal bovine serum and bovine serum albumin on in vitro production of cattle embryos. In: ANNUAL CONFERENCE OT THE INTERNATIONAL EMBRYO TRANSFER SOCIETY, 36., 2010, Cordoba. Proceedings... Córdoba: ITES, 2010; Reproduction, Fertility and development, v. 22, n. 1, p. 197, 2010.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 11. | | SARAIVA, N. Z.; OLIVEIRA, C. S.; TETZNER, T. A. D.; LIMA, M. R. de; NICIURA, S. C. M.; GARCIA, J. M. Effects of demecolcine on the meiotic cell-cycle and microtubular kinetics of activated bovine oocytes submitted to chemical enucleation. In: ANUAL CONFERENCE OF INTERNATIONAL EMBRYO TRANSFER SOCIETY, 24., Phoenix, Arizona. Phoenix: CSIRO, 2012 p. 119| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 12. | | SARAIVA, N. Z.; PERECIN, F.; MÉO, S. C.; FERREIRA, C. R.; TETZNER, T. A. D.; OLIVEIRA, J. C.; GARCIA, J. M. Effects of demecolcine on microtubule composition and chemically assisted enucleation of bovine oocytes. Reproduction, Fertility and Development, v. 20, n. 1, p. 107-108, 2008.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 13. | | TETZNER, T. A. D.; SARAIVA, N. Z.; PERECIN, F.; OLIVEIRA, C. S.; MONTEIRO, F. M.; LIMA, M. R.; NICIURA, S. C. M.; FERREIRA, C. R.; GARCIA, J. M. Effects of embryonic fluid and serum replacer as protein sources for in vitro maturation of bovine oocytes. In: INTERNATIONAL SYMPOSIUM ANIMAL BIOLOGY OF REPRODUCTIVE, 3., 2010, Águas de São Pedro: CBRA: USP/FMVZ, 2010| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 14. | | TETZNER, T. A. D.; SARAIVA, N. Z.; PERECIN, F.; NICIURA, S. C. M.; FERREIRA, C. R.; OLIVEIRA, C. S.; GARCIA, J. M. The effects of ovalbumin as a protein source during the in vitro production of bovine embryos. Revista Brasileira de Zootecnia, v. 40, n. 10, p. 2135-2141, oct. 2011.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 15. | | SARAIVA, N. Z.; OLIVEIRA, C. S.; TETZNER, T. A. D.; LIMA, M. R. de; NICIURA, S. C. M.; GARCIA, J. M. Expressão dos genes xist, g6pd e hspa1 em blastocistos bovinos reconstituídos por tn a partir de oócitos receptores produzidos por enucleação assistida quimicamente. In: REUNIÃO ANUAL DA SOCIEDADE BRASILEIRA DE TECNOLOGIA DE EMBRIÕES, 25., 2011, Cumbuco. O impacto das biotecnologias reprodutivas na saúde e produção animal - anais. Cumbuco: SBTE, 2011. p. 437| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 16. | | MEO, S. C.; FERREIRA, C. R.; PERECIN, F.; SARAIVA, N. Z.; TETZNER, T. A. D.; YAMAZAKI, W.; LEA, C. L. V.; MEIRELLES, F. V.; GARCIA, J. M. Karyoplast exchange between strontium- and 6-DMAP-parthenogenetically activated zygotes of cattle. Animal Reproduction Science, v. 116, n. 3-4, p. 381-385, dec. 2009.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 2 |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 17. | | SARAIVA, N. Z.; OLIVEIRA, C. S.; TETZNER, T. A. D.; SOUZA, M. M.; LIMA, M. R.; NICIURA, S. C. M.; GARCIA, J. M. Métodos alternativos de enucleação oocitária utilizados na transferência nuclear em animais. Revista Brasileira de Reprodução Animal, v. 34, n. 4, p. 197-205, out./dez. 2010.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 3 |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 18. | | TETZNER, T. A. D.; SARAIVA, N. Z.; PERECIN, F.; FERREIRA, C. R.; MÉO, S. C.; OLIVEIRA, C. S.; MELO, D. S.; MONTEIRO, F. M.; VANTINI, R.; GARCIA, J. M. Avaliação da influência da ovalbumina (OVA) como suplemento protéico durante a etapa de fecundação in vitro de oócitos bovinos. Acta Scientiae Veterinariae, v. 35, supl. 3, p. s1181, 2007; In: REUNIÃO ANUAL DA SOCIEDADE BRASILEIRA DE TECNOLOGIA DE EMBRIÕES, 21., 2007, Salvador. Anais... Salvador: UFRGS, 2007.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 19. | | TETZNER, T. A. D.; SARAIVA, N. Z.; PERECIN, F.; FERRERIA, C. R.; NICIURA, S. C. M.; OLIVEIRA, C. MELO, D. S.; MELO, D. S.; MONTEIRO, F. M.; VANTINI, R.; GARCIA, J. M. Avaliação da qualidade em embriões PIV com a substituição do soro fetal bovino e da albumina sérica bovina pela ovalbumina. Acta Scientiae Veterinariae, v. 36, (Supl. 2), 2008.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| 20. | | TETZNER, T. A. D.; SARAIVA, N. Z.; PERECIN, F.; FERREIRA, C. R.; MÉO, S. C.; OLIVEIRA, C. S.; MELO, D. S.; MONTEIRO, F. M.; VANTINI, R.; GARCIA, J. M. Efeitos da substituição do soro fetal bovino (SFB) e da albumina sérica bovina (BSA) pela ovalbumina (OVA) na produção in vitro de embriões bovinos. Acta Scientiae Veterinariae, v. 35, supl. 3, p. s1181, 2007; In: REUNIÃO ANUAL DA SOCIEDADE BRASILEIRA DE TECNOLOGIA DE EMBRIÕES, 21., 2007, Salvador. Anais... Salvador: UFRGS, 2007.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Pecuária Sudeste. |
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| Registros recuperados : 24 | |
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| Nenhum registro encontrado para a expressão de busca informada. |
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