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Registro Completo |
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Biblioteca(s): |
Embrapa Pecuária Sudeste. |
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Data corrente: |
24/03/2008 |
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Data da última atualização: |
22/06/2023 |
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Tipo da produção científica: |
Resumo em Anais de Congresso |
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Autoria: |
SARAIVA, N. Z.; PERECIN, F.; MÉO, S. C.; FERREIRA, C. R.; TETZNER, T. A. D.; OLIVEIRA, J. C.; GARCIA, J. M. |
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Afiliação: |
Naiara Z. Saraiva, UNESP/Jaboticabal; Felipe Perecin USP/Pirassununga Simone Crisitina Méo CPPSE; Christina Ramires Ferreira, Unicamp; Tatiane Almeida D. Tetzner, UNESP/Jaboticabal; J. M. Oliveira, UNESP/Jaboticabal; Joaquim Mansano Garcia, UNESP/Jaboticabal. |
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Título: |
Effects of demecolcine on microtubule composition and chemically assisted enucleation of bovine oocytes. |
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Ano de publicação: |
2008 |
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Fonte/Imprenta: |
Reproduction, Fertility and Development, v. 20, n. 1, p. 107-108, 2008. |
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Idioma: |
Inglês |
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Conteúdo: |
The developmental competence of enucleated oocytes is a key factor that determines the overall success of animal cloning. Enucleation is an invasive procedure in traditional nuclear transfer (NT). The objective of this work was to evaluate the effects of demecolcine, a microtubule-depolymerizing agent, on metaphase II (MII) bovine oocytes and to verify the capacity of embryonic development after NT using chemically assisted enucleation. In the first experiment, oocytes after 21 h of IVM were exposed for 2 h to several concentrations of demecolcine: 0 (control), 0.025, 0.05, 0.2, and 0.4 µg mL?1, and evaluated in relation to membrane protrusion formation. After the best concentration of demecolcine was determined, the nuclear and microtubular dynamics of the treated oocytes were evaluated by immunofluorescence microscopy of tubulin and chromatin (Liu et al. 1998 Biol. Reprod. 5, 537?545) in a second experiment. The results were analyzed by Duncan and Tukey tests in Experiments I and II, respectively, and a level of 5% significance was used. In Experiment III, embryonic development following NT was determined using adult donor cells, derived from a skin biopsy. After 19 h of IVM, oocytes were exposed to demecolcine (0.05 µg mL?1) for 2 h, and enucleation was performed in oocytes that presented membrane protrusions. Samples of cytoplasts were stained with Hoechst 33342 for 10 min (10 µg mL?1) for evaluation of enucleation effectiveness. The remaining oocytes were reconstituted by NT, fused (two pulses of 2.0 kV cm?1 for 20 µs each, in 0.28 m mannitol solution), chemically activated (5 mm ionomycin for 5 min and 2 mm 6-DMAP for 4 h), and cultured in SOF medium with 2.5% FCS and 0.5% BSA. The 0.05 µg mL?1 concentration resulted in greater protrusion rates (55.1%; 211/388) in comparison to 0, 0.025, 0.2, and 0.4 µg mL?1 concentrations (0, 42.6, 45.1, and 39.3%, respectively). In Experiment II, at the beginning of treatment, the majority of the oocytes were in MII (40.7%) or anaphase I/telophase I (AI/TI; 22.4%). Effects of demecolcine occurred within only 0.5 h of treatment, with a significant increase in oocytes with complete depletion of microtubules (21.8%; initial average: 1.9%) and a reduction in the proportion at MII (13.5%) and AI/TI (8.2%). New polymerization of microtubules was observed when treated oocytes were then cultured in drug-free medium for 6 h (42.4% oocytes with two evident sets of microtubules). In Experiment III, we verified the effectiveness of the chemically assisted enucleation technique (90.6%; 77/85). We evaluated 515 oocytes, of which 219 (42.5%) had protrusions and were enucleated. After losses in the NT procedure and fusion, 58 reconstructed 1-cells were cultured resulting in cleavage and blastocyst rates of 84.5% (49/58) and 27.6% (16/58), respectively, with great variation in blastocyst production between the three replicates (12.5% to 47%). In conclusion, demecolcine can be used at lower concentrations than those used routinely, and the chemically assisted enucleation method has proven highly efficient and does not appear to inhibit embryonic development in the bovine MenosThe developmental competence of enucleated oocytes is a key factor that determines the overall success of animal cloning. Enucleation is an invasive procedure in traditional nuclear transfer (NT). The objective of this work was to evaluate the effects of demecolcine, a microtubule-depolymerizing agent, on metaphase II (MII) bovine oocytes and to verify the capacity of embryonic development after NT using chemically assisted enucleation. In the first experiment, oocytes after 21 h of IVM were exposed for 2 h to several concentrations of demecolcine: 0 (control), 0.025, 0.05, 0.2, and 0.4 µg mL?1, and evaluated in relation to membrane protrusion formation. After the best concentration of demecolcine was determined, the nuclear and microtubular dynamics of the treated oocytes were evaluated by immunofluorescence microscopy of tubulin and chromatin (Liu et al. 1998 Biol. Reprod. 5, 537?545) in a second experiment. The results were analyzed by Duncan and Tukey tests in Experiments I and II, respectively, and a level of 5% significance was used. In Experiment III, embryonic development following NT was determined using adult donor cells, derived from a skin biopsy. After 19 h of IVM, oocytes were exposed to demecolcine (0.05 µg mL?1) for 2 h, and enucleation was performed in oocytes that presented membrane protrusions. Samples of cytoplasts were stained with Hoechst 33342 for 10 min (10 µg mL?1) for evaluation of enucleation effectiveness. The remaining oocytes were reconstituted ... Mostrar Tudo |
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Thesagro: |
BOVINO. |
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Thesaurus Nal: |
oocytes. |
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Categoria do assunto: |
-- |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/42090/1/PROCI-2008.00005.pdf
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Marc: |
LEADER 03789nam a2200205 a 4500
001 1048309
005 2023-06-22
008 2008 bl uuuu u00u1 u #d
100 1 $aSARAIVA, N. Z.
245 $aEffects of demecolcine on microtubule composition and chemically assisted enucleation of bovine oocytes.$h[electronic resource]
260 $aReproduction, Fertility and Development, v. 20, n. 1, p. 107-108$c2008
520 $aThe developmental competence of enucleated oocytes is a key factor that determines the overall success of animal cloning. Enucleation is an invasive procedure in traditional nuclear transfer (NT). The objective of this work was to evaluate the effects of demecolcine, a microtubule-depolymerizing agent, on metaphase II (MII) bovine oocytes and to verify the capacity of embryonic development after NT using chemically assisted enucleation. In the first experiment, oocytes after 21 h of IVM were exposed for 2 h to several concentrations of demecolcine: 0 (control), 0.025, 0.05, 0.2, and 0.4 µg mL?1, and evaluated in relation to membrane protrusion formation. After the best concentration of demecolcine was determined, the nuclear and microtubular dynamics of the treated oocytes were evaluated by immunofluorescence microscopy of tubulin and chromatin (Liu et al. 1998 Biol. Reprod. 5, 537?545) in a second experiment. The results were analyzed by Duncan and Tukey tests in Experiments I and II, respectively, and a level of 5% significance was used. In Experiment III, embryonic development following NT was determined using adult donor cells, derived from a skin biopsy. After 19 h of IVM, oocytes were exposed to demecolcine (0.05 µg mL?1) for 2 h, and enucleation was performed in oocytes that presented membrane protrusions. Samples of cytoplasts were stained with Hoechst 33342 for 10 min (10 µg mL?1) for evaluation of enucleation effectiveness. The remaining oocytes were reconstituted by NT, fused (two pulses of 2.0 kV cm?1 for 20 µs each, in 0.28 m mannitol solution), chemically activated (5 mm ionomycin for 5 min and 2 mm 6-DMAP for 4 h), and cultured in SOF medium with 2.5% FCS and 0.5% BSA. The 0.05 µg mL?1 concentration resulted in greater protrusion rates (55.1%; 211/388) in comparison to 0, 0.025, 0.2, and 0.4 µg mL?1 concentrations (0, 42.6, 45.1, and 39.3%, respectively). In Experiment II, at the beginning of treatment, the majority of the oocytes were in MII (40.7%) or anaphase I/telophase I (AI/TI; 22.4%). Effects of demecolcine occurred within only 0.5 h of treatment, with a significant increase in oocytes with complete depletion of microtubules (21.8%; initial average: 1.9%) and a reduction in the proportion at MII (13.5%) and AI/TI (8.2%). New polymerization of microtubules was observed when treated oocytes were then cultured in drug-free medium for 6 h (42.4% oocytes with two evident sets of microtubules). In Experiment III, we verified the effectiveness of the chemically assisted enucleation technique (90.6%; 77/85). We evaluated 515 oocytes, of which 219 (42.5%) had protrusions and were enucleated. After losses in the NT procedure and fusion, 58 reconstructed 1-cells were cultured resulting in cleavage and blastocyst rates of 84.5% (49/58) and 27.6% (16/58), respectively, with great variation in blastocyst production between the three replicates (12.5% to 47%). In conclusion, demecolcine can be used at lower concentrations than those used routinely, and the chemically assisted enucleation method has proven highly efficient and does not appear to inhibit embryonic development in the bovine
650 $aoocytes
650 $aBOVINO
700 1 $aPERECIN, F.
700 1 $aMÉO, S. C.
700 1 $aFERREIRA, C. R.
700 1 $aTETZNER, T. A. D.
700 1 $aOLIVEIRA, J. C.
700 1 $aGARCIA, J. M.
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Registro original: |
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. |
| |
| 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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