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Registro Completo |
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Biblioteca(s): |
Embrapa Amazônia Oriental. |
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Data corrente: |
27/10/2015 |
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Data da última atualização: |
30/05/2022 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
SARAIVA, N. Z.; OLIVEIRA, C. S.; LEAL, C. L. V.; LIMA, M. R. de; CALLADO, M. del; VANTINI, R.; MONTEIRO, F. M.; NICIURA, S. C. M.; GARCIA, J. M. |
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Afiliação: |
NAIARA ZOCCAL SARAIVA, CPATU; CLARA SLADE OLIVEIRA, CNPGL; Cláudia Lima Verde Leal, USP; Marina Ragagnin de Lima, UNESP; Maite Del Collado, UNESP / USP; Roberta Vantini, UNESP; Fabio Morato Monteiro, Centro APTA Bovinos de Corte, Instituto de Zootecnia; SIMONE CRISTINA MEO NICIURA, CPPSE; Joaquim Mansano Garcia, UNESP. |
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Título: |
Chemically induced enucleation of activated bovine oocytes: chromatin and microtubule organization and production of viable cytoplasts. |
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Ano de publicação: |
2015 |
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Fonte/Imprenta: |
Zygote, v. 23, n. 6, p. 852-862, Dec. 2015. |
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DOI: |
http://dx.doi.org/10.1017/S0967199414000537 |
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Idioma: |
Inglês |
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Conteúdo: |
As the standard enucleation method in mammalian nuclear transfer is invasive and damaging to cytoplast spatial organization, alternative procedures have been developed over recent years. Among these techniques, chemically induced enucleation (IE) is especially interesting because it does not employ ultraviolet light and reduces the amount of cytoplasm eliminated during the procedure. The objective of this study was to optimize the culture conditions with demecolcine of pre-activated bovine oocytes for chemically IE, and to evaluate nuclear and microtubule organization in cytoplasts obtained by this technique and their viability. In the first experiment, a negative effect on oocyte activation was verified when demecolcine was added at the beginning of the process, reducing activation rates by approximately 30%. This effect was not observed when demecolcine was added to the medium after 1.5 h of activation. In the second experiment, although a reduction in the number of microtubules was observed in most oocytes, these structures did not disappear completely during assessment. Approximately 50% of treated oocytes presented microtubule reduction at the end of the evaluation period, while 23% of oocytes were observed to exhibit the complete disappearance of these structures and 28% exhibited visible microtubules. These findings indicated the lack of immediate microtubule repolymerization after culture in demecolcine-free medium, a fact that may negatively influence embryonic development. However, cleavage rates of 63.6?70.0% and blastocyst yield of 15.5?24.2% were obtained in the final experiment, without significant differences between techniques, indicating that chemically induced enucleation produces normal embryos. MenosAs the standard enucleation method in mammalian nuclear transfer is invasive and damaging to cytoplast spatial organization, alternative procedures have been developed over recent years. Among these techniques, chemically induced enucleation (IE) is especially interesting because it does not employ ultraviolet light and reduces the amount of cytoplasm eliminated during the procedure. The objective of this study was to optimize the culture conditions with demecolcine of pre-activated bovine oocytes for chemically IE, and to evaluate nuclear and microtubule organization in cytoplasts obtained by this technique and their viability. In the first experiment, a negative effect on oocyte activation was verified when demecolcine was added at the beginning of the process, reducing activation rates by approximately 30%. This effect was not observed when demecolcine was added to the medium after 1.5 h of activation. In the second experiment, although a reduction in the number of microtubules was observed in most oocytes, these structures did not disappear completely during assessment. Approximately 50% of treated oocytes presented microtubule reduction at the end of the evaluation period, while 23% of oocytes were observed to exhibit the complete disappearance of these structures and 28% exhibited visible microtubules. These findings indicated the lack of immediate microtubule repolymerization after culture in demecolcine-free medium, a fact that may negatively influence embryonic deve... Mostrar Tudo |
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Palavras-Chave: |
Bovine; Chemically induced enucleation; Microtubule; Nuclear transfer. |
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Thesaurus Nal: |
chromatin. |
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Categoria do assunto: |
L Ciência Animal e Produtos de Origem Animal |
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Marc: |
LEADER 02641naa a2200289 a 4500
001 2027313
005 2022-05-30
008 2015 bl uuuu u00u1 u #d
024 7 $ahttp://dx.doi.org/10.1017/S0967199414000537$2DOI
100 1 $aSARAIVA, N. Z.
245 $aChemically induced enucleation of activated bovine oocytes$bchromatin and microtubule organization and production of viable cytoplasts.$h[electronic resource]
260 $c2015
520 $aAs the standard enucleation method in mammalian nuclear transfer is invasive and damaging to cytoplast spatial organization, alternative procedures have been developed over recent years. Among these techniques, chemically induced enucleation (IE) is especially interesting because it does not employ ultraviolet light and reduces the amount of cytoplasm eliminated during the procedure. The objective of this study was to optimize the culture conditions with demecolcine of pre-activated bovine oocytes for chemically IE, and to evaluate nuclear and microtubule organization in cytoplasts obtained by this technique and their viability. In the first experiment, a negative effect on oocyte activation was verified when demecolcine was added at the beginning of the process, reducing activation rates by approximately 30%. This effect was not observed when demecolcine was added to the medium after 1.5 h of activation. In the second experiment, although a reduction in the number of microtubules was observed in most oocytes, these structures did not disappear completely during assessment. Approximately 50% of treated oocytes presented microtubule reduction at the end of the evaluation period, while 23% of oocytes were observed to exhibit the complete disappearance of these structures and 28% exhibited visible microtubules. These findings indicated the lack of immediate microtubule repolymerization after culture in demecolcine-free medium, a fact that may negatively influence embryonic development. However, cleavage rates of 63.6?70.0% and blastocyst yield of 15.5?24.2% were obtained in the final experiment, without significant differences between techniques, indicating that chemically induced enucleation produces normal embryos.
650 $achromatin
653 $aBovine
653 $aChemically induced enucleation
653 $aMicrotubule
653 $aNuclear transfer
700 1 $aOLIVEIRA, C. S.
700 1 $aLEAL, C. L. V.
700 1 $aLIMA, M. R. de
700 1 $aCALLADO, M. del
700 1 $aVANTINI, R.
700 1 $aMONTEIRO, F. M.
700 1 $aNICIURA, S. C. M.
700 1 $aGARCIA, J. M.
773 $tZygote$gv. 23, n. 6, p. 852-862, Dec. 2015.
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Registro original: |
Embrapa Amazônia Oriental (CPATU) |
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