Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B
| Ano de defesa: | 2001 |
|---|---|
| Autor(a) principal: | |
| Orientador(a): | |
| Banca de defesa: | |
| Tipo de documento: | Tese |
| Tipo de acesso: | Acesso aberto |
| dARK ID: | ark:/26339/001300000ns3x |
| Idioma: | por |
| Instituição de defesa: |
Universidade Federal de Santa Maria
Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
| Programa de Pós-Graduação: |
Não Informado pela instituição
|
| Departamento: |
Não Informado pela instituição
|
| País: |
Não Informado pela instituição
|
| Palavras-chave em Português: | |
| Link de acesso: | http://repositorio.ufsm.br/handle/1/26790 |
Resumo: | Experiments were conducted to evaluate the effects of cytochalasin B (CB) on the cryopreservation survival rates of bovine oocytes and embryos. In a first study, the embryonic development of in vitro matured bovine oocytes was evaluated, after exposure to different CB doses associated or not to a centrifugation process. In the experiments I and II, four treatments were applied: centrifugation 2100g for 5 minutes (T1); exposure to 7.5μg/mL CB, for 15 minutes (T2); centrifugation after exposure to CB (T3) and oocytes without CB (T4- control). In the experiment III, the oocytes were submitted to three CB concentrations: 7.5μg/mL (T1); 15μg/mL (T2) and 45μg/mL (T3). The cleavage rates (Exp. I), as well as blastocysts and hatching rates (Exp. II) were not influenced by treatments. The exposure to high doses of CB (Exp. III) didn't reduce the cleavage and blastocysts rates. In the subsequent study, the influence of different concentrations of CB was evaluated in the vitrification process of bovine oocytes and embryos produced in vitro. In the experiment I, oocytes were matured for 22 hours, and then vitrified immediately (Vitri treatment) or exposed for 15 to 20 minutes, to a 7.5μg/mL CB solution (CB7.5Vitri treatment) or 45μg/mL (CB45Vitri treatment) prior to vitrification. After 30 seconds of exposure to the equilibrium solution (SV1) and 20 seconds to the vitrification solution (SV2), the oocytes were vitrified in open pulled straws (OPS). The re-warming process was carried out in two steps of 5 minutes each, in solutions with decreasing concentrations of threalose. No differences were observed (P> 0.05) in the cleavage and embryo rates among Vitri, Cito7.5Vitri and Cito45Vitri treatments, which were lower (P <0.05) than the Control. In the Experiment II, expanded blastocysts were allocated in 3 treatments. In the Vitri treatment, the embryos were exposed for 1 minute to SV1 and 20 seconds to SV2 solution, in which the threalose was replaced by TCM-Hepes. In the CB45Vitri treatment, the vitrification was performed after 10 minutes exposure to 45μg/mL CB solution. No difference was observed (P>0.05) in the re-expansion and hatching rates among Vitri and CB45Vitri groups, which were lower (P<0.05) than Control group. The results of this study indicate that the exposure to cytochalasin B had no harmful effect on bovine oocytes during culture and do not show beneficial effects in the vitrification procedure. |
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Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina BCryopreservation and in vitro development of bovine oocytes treated with cytochalasin BFisiopatologia animalOócito bovinoFisiopatologia da reproduçãoEmbriologia animalEmbriõesReprodução animalCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAExperiments were conducted to evaluate the effects of cytochalasin B (CB) on the cryopreservation survival rates of bovine oocytes and embryos. In a first study, the embryonic development of in vitro matured bovine oocytes was evaluated, after exposure to different CB doses associated or not to a centrifugation process. In the experiments I and II, four treatments were applied: centrifugation 2100g for 5 minutes (T1); exposure to 7.5μg/mL CB, for 15 minutes (T2); centrifugation after exposure to CB (T3) and oocytes without CB (T4- control). In the experiment III, the oocytes were submitted to three CB concentrations: 7.5μg/mL (T1); 15μg/mL (T2) and 45μg/mL (T3). The cleavage rates (Exp. I), as well as blastocysts and hatching rates (Exp. II) were not influenced by treatments. The exposure to high doses of CB (Exp. III) didn't reduce the cleavage and blastocysts rates. In the subsequent study, the influence of different concentrations of CB was evaluated in the vitrification process of bovine oocytes and embryos produced in vitro. In the experiment I, oocytes were matured for 22 hours, and then vitrified immediately (Vitri treatment) or exposed for 15 to 20 minutes, to a 7.5μg/mL CB solution (CB7.5Vitri treatment) or 45μg/mL (CB45Vitri treatment) prior to vitrification. After 30 seconds of exposure to the equilibrium solution (SV1) and 20 seconds to the vitrification solution (SV2), the oocytes were vitrified in open pulled straws (OPS). The re-warming process was carried out in two steps of 5 minutes each, in solutions with decreasing concentrations of threalose. No differences were observed (P> 0.05) in the cleavage and embryo rates among Vitri, Cito7.5Vitri and Cito45Vitri treatments, which were lower (P <0.05) than the Control. In the Experiment II, expanded blastocysts were allocated in 3 treatments. In the Vitri treatment, the embryos were exposed for 1 minute to SV1 and 20 seconds to SV2 solution, in which the threalose was replaced by TCM-Hepes. In the CB45Vitri treatment, the vitrification was performed after 10 minutes exposure to 45μg/mL CB solution. No difference was observed (P>0.05) in the re-expansion and hatching rates among Vitri and CB45Vitri groups, which were lower (P<0.05) than Control group. The results of this study indicate that the exposure to cytochalasin B had no harmful effect on bovine oocytes during culture and do not show beneficial effects in the vitrification procedure.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqO progresso obtido em biotécnicas da reprodução tem levado a novos desafios em muitas áreas. Entre estes se destaca o desenvolvimento de uma técnica eficiente para a criopreservação de oócitos bovinos. Com este objetivo, dois estudos foram conduzidos para avaliar a influência da citocalasina B (CB) na vitrificação de oócitos e embriões bovinos produzidos in vitro. No primeiro, avaliou-se em 3 experimentos (Capítulo 1- Experimentos I, II e III), o desenvolvimento embrionário de oócitos bovinos maturados in vitro, expostos a diferentes concentrações de CB, associada ou não à centrifugação. Nos experimentos I e II, foram conduzidos quatro tratamentos: Centrifugação a 2100g por 5 minutos (T1); Exposição a 7,5μg/mL CB, por 15 minutos (T2); Centrifugação após exposição à CB (T3) e Oócitos controle (T4). No experimento III, os oócitos foram submetidos a três concentrações de CB: 7,5μg/mL (T1); 15μg/mL (T2) e 45μg/mL (T3). A clivagem (Experimento I), bem como a formação e eclosão de blastocistos (Experimento II) não foram influenciadas pelos tratamentos efetuados. A exposição a doses mais elevadas de CB (Experimento III) não reduziu a taxa de clivagem e de blastocistos. Em seqüência, avaliou-se a influência de diferentes concentrações de CB na vitrificação de oócitos e embriões bovinos produzidos in vitro. No capítulo 2 (Experimento I), oócitos foram maturados por 22 horas, sendo imediatamente vitrificados (tratamento Vitri) ou expostos por 15 a 20 minutos, à solução com 7,5μg/mL (tratamento CB7,5Vitri) ou 45μg/mL (tratamento CB45Vitri) de citocalasina B, antes da vitrificação. Após 30 segundos de exposição a solução de equilíbrio (SV1) e 20 segundos a solução de vitrificação (SV2), os oócitos foram vitrificados em palhetas estiradas (OPS). O reaquecimento foi realizado em duas etapas de 5 minutos cada, em soluções decrescentes de trealose. Não foram observadas diferenças (P>0,05) nos percentuais de clivagem e de embriões entre os tratamentos Vitri, Cito7,5Vitri e Cito45Vitri, que foram inferiores (P<0,05) ao controle. No Experimento II, 269 blastocistos expandidos foram distribuídos em 3 tratamentos. No tratamento Vitri os embriões foram expostos por 1 minuto a SV1 e 20 segundos a SV2, que teve a trealose substituida por TCM-Hepes. No tratamento CB45Vitri, a vitrificação ocorreu após exposição por 10 minutos a solução com 45μg/mL de CB. Não foram observadas diferenças (P>0,05) no percentual de re-expansão e eclosão entre os grupos Vitri e CB45Vitri, os quais foram inferiores (P<0,05) ao grupo controle. Os resultados indicam que a exposição a citocalasina B não reduz a viabilidade de oócitos bovinos durante o cultivo e não produz efeito benéfico no processo de vitrificação.Universidade Federal de Santa MariaBrasilMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaCentro de Ciências RuraisRubin, Mara Iolanda Batistellahttp://lattes.cnpq.br/5140943832465284Bernardi, Mari LourdesPegoraro, Ligia Margareth CantarelliAzambuja, Ricardo Marques deDe La Côrte, Flávio DesessardsMezzalira, Alceu2022-11-08T12:14:36Z2022-11-08T12:14:36Z2001-07-20info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/26790ark:/26339/001300000ns3xporAttribution-NonCommercial-NoDerivatives 4.0 Internationalinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-11-08T12:14:37Zoai:repositorio.ufsm.br:1/26790Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/PUBhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.com||manancial@ufsm.bropendoar:2022-11-08T12:14:37Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
| dc.title.none.fl_str_mv |
Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B Cryopreservation and in vitro development of bovine oocytes treated with cytochalasin B |
| title |
Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B |
| spellingShingle |
Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B Mezzalira, Alceu Fisiopatologia animal Oócito bovino Fisiopatologia da reprodução Embriologia animal Embriões Reprodução animal CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
| title_short |
Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B |
| title_full |
Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B |
| title_fullStr |
Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B |
| title_full_unstemmed |
Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B |
| title_sort |
Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B |
| author |
Mezzalira, Alceu |
| author_facet |
Mezzalira, Alceu |
| author_role |
author |
| dc.contributor.none.fl_str_mv |
Rubin, Mara Iolanda Batistella http://lattes.cnpq.br/5140943832465284 Bernardi, Mari Lourdes Pegoraro, Ligia Margareth Cantarelli Azambuja, Ricardo Marques de De La Côrte, Flávio Desessards |
| dc.contributor.author.fl_str_mv |
Mezzalira, Alceu |
| dc.subject.por.fl_str_mv |
Fisiopatologia animal Oócito bovino Fisiopatologia da reprodução Embriologia animal Embriões Reprodução animal CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
| topic |
Fisiopatologia animal Oócito bovino Fisiopatologia da reprodução Embriologia animal Embriões Reprodução animal CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
| description |
Experiments were conducted to evaluate the effects of cytochalasin B (CB) on the cryopreservation survival rates of bovine oocytes and embryos. In a first study, the embryonic development of in vitro matured bovine oocytes was evaluated, after exposure to different CB doses associated or not to a centrifugation process. In the experiments I and II, four treatments were applied: centrifugation 2100g for 5 minutes (T1); exposure to 7.5μg/mL CB, for 15 minutes (T2); centrifugation after exposure to CB (T3) and oocytes without CB (T4- control). In the experiment III, the oocytes were submitted to three CB concentrations: 7.5μg/mL (T1); 15μg/mL (T2) and 45μg/mL (T3). The cleavage rates (Exp. I), as well as blastocysts and hatching rates (Exp. II) were not influenced by treatments. The exposure to high doses of CB (Exp. III) didn't reduce the cleavage and blastocysts rates. In the subsequent study, the influence of different concentrations of CB was evaluated in the vitrification process of bovine oocytes and embryos produced in vitro. In the experiment I, oocytes were matured for 22 hours, and then vitrified immediately (Vitri treatment) or exposed for 15 to 20 minutes, to a 7.5μg/mL CB solution (CB7.5Vitri treatment) or 45μg/mL (CB45Vitri treatment) prior to vitrification. After 30 seconds of exposure to the equilibrium solution (SV1) and 20 seconds to the vitrification solution (SV2), the oocytes were vitrified in open pulled straws (OPS). The re-warming process was carried out in two steps of 5 minutes each, in solutions with decreasing concentrations of threalose. No differences were observed (P> 0.05) in the cleavage and embryo rates among Vitri, Cito7.5Vitri and Cito45Vitri treatments, which were lower (P <0.05) than the Control. In the Experiment II, expanded blastocysts were allocated in 3 treatments. In the Vitri treatment, the embryos were exposed for 1 minute to SV1 and 20 seconds to SV2 solution, in which the threalose was replaced by TCM-Hepes. In the CB45Vitri treatment, the vitrification was performed after 10 minutes exposure to 45μg/mL CB solution. No difference was observed (P>0.05) in the re-expansion and hatching rates among Vitri and CB45Vitri groups, which were lower (P<0.05) than Control group. The results of this study indicate that the exposure to cytochalasin B had no harmful effect on bovine oocytes during culture and do not show beneficial effects in the vitrification procedure. |
| publishDate |
2001 |
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2001-07-20 2022-11-08T12:14:36Z 2022-11-08T12:14:36Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/doctoralThesis |
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ark:/26339/001300000ns3x |
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por |
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Attribution-NonCommercial-NoDerivatives 4.0 International |
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Universidade Federal de Santa Maria Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
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Universidade Federal de Santa Maria Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
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reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
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Universidade Federal de Santa Maria (UFSM) |
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Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
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