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Registro Completo |
Biblioteca(s): |
Embrapa Pecuária Sudeste. |
Data corrente: |
24/03/2008 |
Data da última atualização: |
22/06/2023 |
Tipo da produção científica: |
Resumo em Anais de Congresso |
Autoria: |
SARAIVA, N. Z.; PERECIN, F.; MÉO, S. C.; FERREIRA, C. R.; TETZNER, T. A. D.; OLIVEIRA, J. C.; GARCIA, J. M. |
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. |
Título: |
Effects of demecolcine on microtubule composition and chemically assisted enucleation of bovine oocytes. |
Ano de publicação: |
2008 |
Fonte/Imprenta: |
Reproduction, Fertility and Development, v. 20, n. 1, p. 107-108, 2008. |
Idioma: |
Inglês |
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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BOVINO. |
Thesaurus Nal: |
oocytes. |
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 03796nam 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 $aReproduction, Fertility and Development, v. 20, n. 1, p. 107-108, 2008.$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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Embrapa Pecuária Sudeste (CPPSE) |
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Registros recuperados : 19 | |
1. | | PIRES, R. M. L.; PIRES, R. L.; MÉO, S. C. Quimerismo 60, xx/60, xy e freemartismo em bovinos nascidos de partos gemelares heterossexuais. Boletim de Indústria Animal, Nova Odessa, v. 66, n. 1, p. 45-55, jun./mar. 2009.Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 4 |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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2. | | 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. |
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3. | | CARDOSO, D.; CURCI, V. C. M.; NOGUEIRA, A. H. de C.; VERÍSSIMO, C. J.; MÉO, S. C.; MOLENTO, M. B. Situação da resitência de helmintos de ovinos a anti-helmínticos na região de Araçatuba, Estado de São Paulo. In: CONGRESSO BRASILEIRO DE ZOOTECNIA, 19., 2009, Águas de Lindóia. Anais... Águas de Lindóia: FZEA/USP: ABZ, 2009Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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4. | | 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. |
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5. | | MÉO, S. C.; YAMAZAKI, W.; FERREIRA, C. R.; PERECIN, F.; SARAIVA, N. Z.; LEAL, C. L. V.; GARCIA, J. M. Parthenogenetic activation of bovine oocytes using single and combined strontium, ionomycin and 6-dimethylaminopurine treatments. Zygote, v. 15, p. 295-306, nov. 2007.Tipo: Artigo em Periódico Indexado | Circulação/Nível: Internacional - A |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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6. | | MELLO, S. S.; MÉO, S. C.; GROMBONI, J. G. G.; ROCHA, M. I. P.; VENERONI, G. B. Validação dos resultados de genotipagem por Taqman por meio de sequenciamento de genes relacionados à qualidade da carne bovina. In: CONGRESSO DE INICIAÇÃO CIENTIFICA, 18., 2010, São Carlos, SP. Anais ... São Carlos: UFSCar, 2010.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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7. | | MÉO, S. C.; FERREIRA, C. R.; CHIARATTI, M. R.; MEIRELLES, F. V.; REGITANO, L. C. de A.; ALENCAR, M. M. de; BARBOSA, P. F. Characterization of mitochondrial genotypes in the foundation herd of the Canchim beef cattle breed. Genetics and Molecular Research, v. 8, n. 1, p. 261-267, 2009.Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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8. | | 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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9. | | SARAIVA, N. Z.; OLIVEIRA, C. S.; COLLADO, M. del; LIMA, M. R.; MEO, S. C.; FIGUEIRO, M. R.; VANTINI, R.; GARCIA, J. M. Effects of cAMP modulators during in vitro prematuration of bovine oocytes on gap junctional communication and embryo development potential. Animal Reproduction, v. 13, n. 3, p. 602, Jul./Sep. 2016.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Amazônia Oriental. |
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10. | | 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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11. | | FERREIRA, C. R.; BURGSTALLER, J. P.; PERECIN, F.; GARCIA, J. M.; CHIARATTI, M. R.; MÉO, S. C.; MULLER, M.; SMITH, L. C.; MEIRELLES, F. V.; STEINBORN, R. Pronounced segregation of donor mitochondria introduced by bovine Ooplasmic tranfer to the female germ-line. Biology of Reproduction, v. 82, n. 03, p. 563-571, mar. 2010.Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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12. | | 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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13. | | MÉO, S. C.; TIZIOTO, P. C.; SIQUEIRA, F.; ROSA, A. do N.; SILVA, L. O. C. da; TORRES JUNIOR, R. A. de A.; VENERONI, G. B.; IBELLI, A. M. G.; ALENCAR, M. M. de; REGITANO, L. C. de A. Caracterização do haplótipo mitocondrial de uma amostra de touros da raça Nelore, representativa das principais linhagens comercializadas no Brasil. In: WORKSHOP DA REDE GENÔMICA ANIMAL, 1., 2009, Fortaleza. Anais... Fortaleza, Rede Genômica Animal, 2009.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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14. | | VERÍSSIMO, C. J.; MEO, S. C.; OLIVEIRA, D. C.; CURCI, V. M.; UENO, T. H.; RODRIGUES, C. C.; COSTA, R. D.; PEREIRA, J. R.; MARGATHO, L. F.; CHIEBAO, D. P.; MOLENTO, M. B. Monitoring the efficacy of anthelmintic drugs in small ruminants in the State of São Paulo, Brazil: preliminary results. In: WORLD ASSOCIATION FOR THE ADVANCEMENT OF VETERINARY PARASITOLOGY, 2009, Calgary. Abstract... Calgary: WAAUP, 2009. p. 40-41.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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15. | | PERECIN, F.; YAMAZAKI, W.; FERREIRA, C. R.; MÉO, S. C.; BIASE, F. H.; MERIGHE, G. K. F.; SARAIVA, N. Z.; TETZNER, T. A. D.; MEIRELLES, F. V.; GARCIA, J. M. Expressão de DNA metiltransferases em blastocistos bovinos produzidos in vivo e in vitro. Acta Scientiae Veterinariae, v. 35, supl. 3, p. 1181, 2007.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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16. | | PERECIN, F.; YAMAZAKI, W.; FERREIRA, C. R.; MÉO, S. C.; BIASE, F. H.; MERIGHE, G. K. F.; SARAIVA, N. Z.; TETZNER, T. A. D.; MEIRELLES, F. V.; GARCIA, J. M. Efeito do protocolo de sincronização celular na produção de clones 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. p. 1267Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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17. | | 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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18. | | CHIARATTI, M. R.; FERREIRA, C. R.; MEIRELLES, F. V.; MÉO, S. C.; PERECIN, F.; SMITH, L. C.; FERRAZ, M. L.; SÁ FILHO, M. F. de; GIMENES, L. U.; BARUSELLI, P. S.; GASPARRINI, B.; GARCIA, J. M. Xenooplasmic transfer between buffalo and bovine enables development of homoplasmic offspring. Cellular Reprogramming, v. 12, n. 2, 2010.Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
Biblioteca(s): Embrapa Pecuária Sudeste. |
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19. | | LARA, M. A. C; RIBEIRO, J. A.; REICHERT, R. H.; PIRES, R. M. L.; ARCARO, J. R. P.; CARVALHO, J. B. P.; DEMARCHI, J. J. A. A.; MÉO, S. C.; BUFARAH, G.; SERENO, J. R. B.; SANTOS, S. A.; ABREU, U. G. P.; MIRANDA-SANTOS, I. K. F.; CONTEL, E. P. B. Relaciones genéticas entre razas bovinas con empleo de marcadores genéticos. In: SIMPÓSIO IBEROAMERICANO SOBRE CONSERVACIÓN Y UTILIZACIÓN DE RECURSOS ZOOGENÉTICOS, 6., 2005, Chiapas, México. Anais. Chiapas, México: CYTED: Universidad Autónoma de Chiapas, 2005. p.159-163.Biblioteca(s): Embrapa Pantanal. |
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Registros recuperados : 19 | |
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