Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFMG |
Texto Completo: | http://hdl.handle.net/1843/33073 |
Resumo: | Oxidative stress is the main cause of low efficiency in oocyte maturation and embryo development in in vitro embryo production, due to an imbalance between the amount of reactive oxygen species (ROS) and antioxidants. The in vitro atmosphere has a high oxygen tension, which associated with other factors such as light interference, presence of sperm and absence of maternal antioxidants lead to a higher production of ROS, when compared to the in vivo atmosphere. The present study was aimed to evaluate the effect of the addition of fullerenol, in distinct concentrations to the in vitro maturation media of bovine oocytes (IVM), on the production rates and quality of the embryos produced. Fullerenol is recognized as a powerful antioxidant, a very electronegative and stable molecule that is able to reach to specific targets. Four distinct IVM media were tested: group 1 – control (CO) (n=461 oocytes): TCM 199 bicarbonate media; group 2 (F1) (n=461): TCM 199 bicarbonate media with 1nM of fullerenol; group 3 (F10) (n=439): TCM 199 bicarbonate media with 10nM of fullerenol; group 4 (F50) (n=451): TCM 199 bicarbonate media with 50nM of fullerenol. Oocytes were matured in incubator during 24 hours at 38oC, 5% of CO2 and humidity of 95%. Subsequently, the oocytes were fertilized and cultivated in the same media of fertilization and cultivation, respectively. On the second day of culture (D2) the cleavage rate was evaluated, and on the seventh day (D7) the blastocyst rate by the total of cleavage embryos and the rate of blastocyst production by the total number of matured oocytes were evaluated. In addition, after observation and definition of those rates, embryos produced were fixed for posterior TUNEL assay. Nuclear maturation was evaluated by meiotic status at the end of oocyte maturation using HOECHST staining. There were no statistical difference (P>0.05) between cleavage rates (73.96%; 73.53%; 73.12%; 76.94%) for CO, F1, F10 and F50, respectively. Blastocyst rate differed between treatments. CO, F1 and F50 showed statistically similar (P>0.05) results (CO=30.15%, F1=27.11%, F50=31.04%). CO and F50 had higher (P<0.05%) blastocyst production than F10 (23.91%), which was equal to F1. The total number of cells per embryo (CO=130.84, F1=129.78, F10=110.72, F50=134.05) and the total number of apoptotic cells (CO=8.52, F1=7.68, F10=7.54, F50=5.85) did not differ (P>0.05) between treatments. The apoptosis rate was higher (P<0.05) in CO (7.43%) than F50 (4.23%). CO, F1 (5.94%) and F10 (6.98%) did not differ (P>0.05), as F1, F10 and F50 were statistically similar (P>0.05). Nuclear maturation (metaphase II) did not differ (P>0.05) between treatments (CO=53.3%; F1=48.2%; F10=48.0%; F50=33.3%). In conclusion, the presence of fullerenol in the IVM media had no effect on nuclear maturation rate and the cleavage rate, while reduced the blastocyst production at 10nM concentration. In high concentration of fullerenol (50nM), cleavage and blastocyst production were similar (P>0.05) to the control group, however reduced apoptosis rate when compared to the same treatment. |
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Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinosEffect of fullerenol addition to the in vitro maturation media of bovine oocytesReprodução animalOocitoBovino embriõesOxidative stress is the main cause of low efficiency in oocyte maturation and embryo development in in vitro embryo production, due to an imbalance between the amount of reactive oxygen species (ROS) and antioxidants. The in vitro atmosphere has a high oxygen tension, which associated with other factors such as light interference, presence of sperm and absence of maternal antioxidants lead to a higher production of ROS, when compared to the in vivo atmosphere. The present study was aimed to evaluate the effect of the addition of fullerenol, in distinct concentrations to the in vitro maturation media of bovine oocytes (IVM), on the production rates and quality of the embryos produced. Fullerenol is recognized as a powerful antioxidant, a very electronegative and stable molecule that is able to reach to specific targets. Four distinct IVM media were tested: group 1 – control (CO) (n=461 oocytes): TCM 199 bicarbonate media; group 2 (F1) (n=461): TCM 199 bicarbonate media with 1nM of fullerenol; group 3 (F10) (n=439): TCM 199 bicarbonate media with 10nM of fullerenol; group 4 (F50) (n=451): TCM 199 bicarbonate media with 50nM of fullerenol. Oocytes were matured in incubator during 24 hours at 38oC, 5% of CO2 and humidity of 95%. Subsequently, the oocytes were fertilized and cultivated in the same media of fertilization and cultivation, respectively. On the second day of culture (D2) the cleavage rate was evaluated, and on the seventh day (D7) the blastocyst rate by the total of cleavage embryos and the rate of blastocyst production by the total number of matured oocytes were evaluated. In addition, after observation and definition of those rates, embryos produced were fixed for posterior TUNEL assay. Nuclear maturation was evaluated by meiotic status at the end of oocyte maturation using HOECHST staining. There were no statistical difference (P>0.05) between cleavage rates (73.96%; 73.53%; 73.12%; 76.94%) for CO, F1, F10 and F50, respectively. Blastocyst rate differed between treatments. CO, F1 and F50 showed statistically similar (P>0.05) results (CO=30.15%, F1=27.11%, F50=31.04%). CO and F50 had higher (P<0.05%) blastocyst production than F10 (23.91%), which was equal to F1. The total number of cells per embryo (CO=130.84, F1=129.78, F10=110.72, F50=134.05) and the total number of apoptotic cells (CO=8.52, F1=7.68, F10=7.54, F50=5.85) did not differ (P>0.05) between treatments. The apoptosis rate was higher (P<0.05) in CO (7.43%) than F50 (4.23%). CO, F1 (5.94%) and F10 (6.98%) did not differ (P>0.05), as F1, F10 and F50 were statistically similar (P>0.05). Nuclear maturation (metaphase II) did not differ (P>0.05) between treatments (CO=53.3%; F1=48.2%; F10=48.0%; F50=33.3%). In conclusion, the presence of fullerenol in the IVM media had no effect on nuclear maturation rate and the cleavage rate, while reduced the blastocyst production at 10nM concentration. In high concentration of fullerenol (50nM), cleavage and blastocyst production were similar (P>0.05) to the control group, however reduced apoptosis rate when compared to the same treatment.O estresse oxidativo é a principal causa de baixa eficiência na maturação de oócitos e no desenvolvimento embrionário in vitro de embriões bovinos. Isso ocorre devido a um desbalanço entre a quantidade de espécies reativas de oxigênio (EROs) e antioxidantes. O ambiente in vitro possui alta tensão de oxigênio que, associada a outros fatores como a interferência da luz, presença de espermatozoide e ausência de antioxidantes maternos, leva à maior produção de EROs, se comparado ao ambiente in vivo. No presente experimento, foi avaliado o efeito da adição de fulerol em diferentes concentrações ao meio de maturação in vitro de oócitos bovinos (MIV) sobre as taxas de produção e qualidade dos embriões produzidos. O fulerol é reconhecido como um potente antioxidante, uma molécula muito eletronegativa, estável, e capaz de alcançar locais específicos. Foram utilizados quatro meios MIV: Grupo 1 – controle (CO) (n=461 oócitos): meio TCM 199 bicarbonato; Grupo 2 (F1) (n=461): meio TCM 199 bicarbonato acrescido de fulerol a 1nM; Grupo 3 (F10) (n=439): meio TCM 199 bicarbonato acrescido de fulerol a 10nM; Grupo 4 (F50) (n=451): meio TCM 199 bicarbonato acrescido de fulerol a 50nM. Os oócitos foram maturados em incubadora por 24 horas a 38,5oC, 5% de CO2 e 95% de umidade. Posteriormente, foram fertilizados e cultivados em mesmo meio de fertilização e cultivo, respectivamente. No segundo dia de cultivo (D2), foi avaliada a taxa de clivagem e no sétimo dia de cultivo (D7) foram avaliadas a taxa de produção de blastocistos sobre os embriões clivados e a taxa de produção de blastocistos sobre o total de oócitos maturados. Além disso, após observação e definição dessas taxas, os embriões produzidos foram fixados para avaliação de apoptose celular pelo método de TUNEL. Os oócitos foram avaliados quanto à maturação nuclear pelo método de Hoechst. Não houve diferença estatística (P>0,05) nas taxas de clivagem (73,96%; 73,53%; 73,12%; 76,94%) para T1; F1; F10 e F50, respectivamente. A produção de blastocistos diferiu (P<0,05) entre os tratamentos. CO, F1 e F50 demonstraram resultados estatisticamente semelhantes (P>0,05) (CO=30,15%; F1=27,11%; F50=31,04%). CO e F50 obtiveram resultados superiores (P<0,05) a F10 (23,91%), que não diferiu (P>0,05) do F1. O número total de células por embrião (CO=130,84; F1=129,78; F10=110,72; F50=134,05) e o número total de células apoptóticas (CO=8,52; F1=7,68; F10=7,54; F50=5,85) não diferiram (P>0,05) entre os tratamentos. A taxa de apoptose foi superior (P<0,05) no CO (7,43%) em relação a F50 (4,23%). CO, F1 (5,94%) e F10 (6,98%) não diferiram (P>0,05), assim como F1, F10 e F50 foram estatisticamente iguais (P>0,05). Não houve diferença (P>0,05) entre a taxa de oócitos maturados (Metáfase II) entre os tratamentos (CO=53,3%; F1=48,2%;F10=48,0%; F50=33,3%). Conclui-se que a presença de fulerol no meio MIV não interferiu nas taxas maturação dos oócitos e de clivagem, porém reduziu a produção de blastocistos em meio na concentração de 10nM de fulerol. Na concentração de 50nM de fulerol os resultados de clivagem e produção de blastocistos foram iguais às do grupo controle, porém reduziu a taxa de apoptose, quando comparado com o esse mesmo grupo.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorUniversidade Federal de Minas GeraisBrasilVET - DEPARTAMENTO DE CLÍNICA E CIRURGIAPrograma de Pós-Graduação em Medicina VeterináriaUFMGÁlan Maia Borgeshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4796232T6&tokenCaptchar=03AERD8XqnjKrFn6ub1rL1esWQR93oo-_DSNX0NtxDq1d3ppgygysyP5vRSeBMeG62m5EeNZQNwbW8PPgT2c18AvqyUjZWB3ulKHxL1gMQ_eeMHLyBgv2M7g74n1onvCOa2DWXxELm0MDxC2yVlTSL_qCJF00rp3Sj-O7Reo1E7OsJI70csvfLn2cFSE6_lpC_aWm-CdO31H90eljdUJEkgoi_-hMzwii3iVYHnifAs9ajdkJ-maFeWi6gC_bltII9xiavjWQaCiYwwEipfCSIlevVv5rflkfHGjrpyCbYVDtUKM3kKRIPhQoNsDN1JOAlRSLWFB5wFh-37fAUePaOKQD-tW8sxH-PVRVQBn0o8hTc0ukAgvNvPHqGvhk0iuC2BSNJX7-w36JY5j_7aBd2Ofm6JvfB11qMGnouH8w-grJNVLADJ4DIQV5baNYafTRh6HXC5o6d9-8uQ7m3iD24t24XQlyEAvLtCwLetícia Zoccolaro OliveiraMariana Machado NevesVictor Mutti Drummond Ribeiro Prata2020-04-01T15:39:25Z2020-04-01T15:39:25Z2019-11-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/1843/33073porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG2020-04-02T06:25:40Zoai:repositorio.ufmg.br:1843/33073Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2020-04-02T06:25:40Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false |
dc.title.none.fl_str_mv |
Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos Effect of fullerenol addition to the in vitro maturation media of bovine oocytes |
title |
Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos |
spellingShingle |
Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos Victor Mutti Drummond Ribeiro Prata Reprodução animal Oocito Bovino embriões |
title_short |
Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos |
title_full |
Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos |
title_fullStr |
Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos |
title_full_unstemmed |
Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos |
title_sort |
Efeito da adição do fulerol ao meio de maturação in vitro de oócitos bovinos |
author |
Victor Mutti Drummond Ribeiro Prata |
author_facet |
Victor Mutti Drummond Ribeiro Prata |
author_role |
author |
dc.contributor.none.fl_str_mv |
Álan Maia Borges http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4796232T6&tokenCaptchar=03AERD8XqnjKrFn6ub1rL1esWQR93oo-_DSNX0NtxDq1d3ppgygysyP5vRSeBMeG62m5EeNZQNwbW8PPgT2c18AvqyUjZWB3ulKHxL1gMQ_eeMHLyBgv2M7g74n1onvCOa2DWXxELm0MDxC2yVlTSL_qCJF00rp3Sj-O7Reo1E7OsJI70csvfLn2cFSE6_lpC_aWm-CdO31H90eljdUJEkgoi_-hMzwii3iVYHnifAs9ajdkJ-maFeWi6gC_bltII9xiavjWQaCiYwwEipfCSIlevVv5rflkfHGjrpyCbYVDtUKM3kKRIPhQoNsDN1JOAlRSLWFB5wFh-37fAUePaOKQD-tW8sxH-PVRVQBn0o8hTc0ukAgvNvPHqGvhk0iuC2BSNJX7-w36JY5j_7aBd2Ofm6JvfB11qMGnouH8w-grJNVLADJ4DIQV5baNYafTRh6HXC5o6d9-8uQ7m3iD24t24XQlyEAvLtCw Letícia Zoccolaro Oliveira Mariana Machado Neves |
dc.contributor.author.fl_str_mv |
Victor Mutti Drummond Ribeiro Prata |
dc.subject.por.fl_str_mv |
Reprodução animal Oocito Bovino embriões |
topic |
Reprodução animal Oocito Bovino embriões |
description |
Oxidative stress is the main cause of low efficiency in oocyte maturation and embryo development in in vitro embryo production, due to an imbalance between the amount of reactive oxygen species (ROS) and antioxidants. The in vitro atmosphere has a high oxygen tension, which associated with other factors such as light interference, presence of sperm and absence of maternal antioxidants lead to a higher production of ROS, when compared to the in vivo atmosphere. The present study was aimed to evaluate the effect of the addition of fullerenol, in distinct concentrations to the in vitro maturation media of bovine oocytes (IVM), on the production rates and quality of the embryos produced. Fullerenol is recognized as a powerful antioxidant, a very electronegative and stable molecule that is able to reach to specific targets. Four distinct IVM media were tested: group 1 – control (CO) (n=461 oocytes): TCM 199 bicarbonate media; group 2 (F1) (n=461): TCM 199 bicarbonate media with 1nM of fullerenol; group 3 (F10) (n=439): TCM 199 bicarbonate media with 10nM of fullerenol; group 4 (F50) (n=451): TCM 199 bicarbonate media with 50nM of fullerenol. Oocytes were matured in incubator during 24 hours at 38oC, 5% of CO2 and humidity of 95%. Subsequently, the oocytes were fertilized and cultivated in the same media of fertilization and cultivation, respectively. On the second day of culture (D2) the cleavage rate was evaluated, and on the seventh day (D7) the blastocyst rate by the total of cleavage embryos and the rate of blastocyst production by the total number of matured oocytes were evaluated. In addition, after observation and definition of those rates, embryos produced were fixed for posterior TUNEL assay. Nuclear maturation was evaluated by meiotic status at the end of oocyte maturation using HOECHST staining. There were no statistical difference (P>0.05) between cleavage rates (73.96%; 73.53%; 73.12%; 76.94%) for CO, F1, F10 and F50, respectively. Blastocyst rate differed between treatments. CO, F1 and F50 showed statistically similar (P>0.05) results (CO=30.15%, F1=27.11%, F50=31.04%). CO and F50 had higher (P<0.05%) blastocyst production than F10 (23.91%), which was equal to F1. The total number of cells per embryo (CO=130.84, F1=129.78, F10=110.72, F50=134.05) and the total number of apoptotic cells (CO=8.52, F1=7.68, F10=7.54, F50=5.85) did not differ (P>0.05) between treatments. The apoptosis rate was higher (P<0.05) in CO (7.43%) than F50 (4.23%). CO, F1 (5.94%) and F10 (6.98%) did not differ (P>0.05), as F1, F10 and F50 were statistically similar (P>0.05). Nuclear maturation (metaphase II) did not differ (P>0.05) between treatments (CO=53.3%; F1=48.2%; F10=48.0%; F50=33.3%). In conclusion, the presence of fullerenol in the IVM media had no effect on nuclear maturation rate and the cleavage rate, while reduced the blastocyst production at 10nM concentration. In high concentration of fullerenol (50nM), cleavage and blastocyst production were similar (P>0.05) to the control group, however reduced apoptosis rate when compared to the same treatment. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-11-01 2020-04-01T15:39:25Z 2020-04-01T15:39:25Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1843/33073 |
url |
http://hdl.handle.net/1843/33073 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais Brasil VET - DEPARTAMENTO DE CLÍNICA E CIRURGIA Programa de Pós-Graduação em Medicina Veterinária UFMG |
publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais Brasil VET - DEPARTAMENTO DE CLÍNICA E CIRURGIA Programa de Pós-Graduação em Medicina Veterinária UFMG |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFMG instname:Universidade Federal de Minas Gerais (UFMG) instacron:UFMG |
instname_str |
Universidade Federal de Minas Gerais (UFMG) |
instacron_str |
UFMG |
institution |
UFMG |
reponame_str |
Repositório Institucional da UFMG |
collection |
Repositório Institucional da UFMG |
repository.name.fl_str_mv |
Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG) |
repository.mail.fl_str_mv |
repositorio@ufmg.br |
_version_ |
1816829638778814464 |