Criopreservação e desenvolvimento de oócitos bovinos tratados com citocalasina B

Detalhes bibliográficos
Ano de defesa: 2001
Autor(a) principal: Mezzalira, Alceu
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
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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spelling 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
dc.date.none.fl_str_mv 2001-07-20
2022-11-08T12:14:36Z
2022-11-08T12:14:36Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/26790
dc.identifier.dark.fl_str_mv ark:/26339/001300000ns3x
url http://repositorio.ufsm.br/handle/1/26790
identifier_str_mv ark:/26339/001300000ns3x
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com||manancial@ufsm.br
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