AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)

Detalhes bibliográficos
Autor(a) principal: Amanda Godoi Navarezi
Data de Publicação: 2022
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFMS
Texto Completo: https://repositorio.ufms.br/handle/123456789/4731
Resumo: The demand for new experimental models grows proportionately as pre-clinical tests need more refined and reproducible results, thus also contributing to one of the main concepts in Laboratory Animal Science: the 3 Rs, which are the reduction of animals in research, refined biomodels and techniques who seek to alleviate stress and discomfort, and replacement of experimental models with validated alternative techniques. An example of a biomodel created in the laboratory is the NOD mice (Nonobese Diabetic), which has contributed mainly to research involving type 1 diabetes mellitus, since the autoimmune process occurs in a very similar way to humans, in addition to several other areas that can be studied from this experimental model. With this in mind, the UFMS Central Animal Facility received NOD couples, which were established in the Breeding Area, where they are produced for supply to the Institution's researchers. With the establishment of this new strain, the present work sought to evaluate the reproduction of NOD mice, mated in the breeding colony, and in obtaining oocytes and embryos with and without hormonal stimulation. Data from 4 births of 20 couples kept in intensive monogamous mating system in the breeding area were used to calculate the productivity indexes. To obtain oocytes and embryos, the Whitten effect was used to synchronize estrus in all females in the study. The hormonal protocol was performed in 20 females through the administration via IP of 7.5IU of PMSG on day 0, and 48 hours later 7.5IU of hCG followed by mating of 10 females with vasectomized males (n=10) to obtain oocytes and the other 10 with intact males (n=10) to obtain embryos. On day 4, the presence of vaginal tampons and oocyte collection were observed. On day 5, embryos were collected. The same mating procedure was performed for the group of females without hormonal stimulation (n=40). In the breeding colony, a litter average of 7.50±0.68 was observed; 5.25±0.95; 2.85±0.88 and 2.20±0.73, from the first to the fourth delivery, respectively. In females that did not receive hormonal stimulation, an average of 6.25±1.15 oocytes and 2.90±0.86 embryos in the 2-cell phase (D2) were obtained, while the superovulated females obtained 79.20±8.43 oocytes and 19.90±7.90 D2 embryos. Regarding the percentage of viable embryos (D2) over the total number of embryos observed, the natural group obtained an average of 20.74±6.27 and the group of superovulated females presented 24.71±6.26. Thus, it was concluded that in intensive monogamous mating, the NOD strain shows a significant drop from the third parturition, and they present a satisfactory response to superovulation in terms of the number of oocytes and embryos, but in relation to the percentage of viable embryos there was no significant difference between groups. Therefore, superovulation is a viable option for obtaining oocytes and embryos with a reduced number of animals, being an important tool for the creation of a germplasm bank and assisted reproduction of the NOD strain. Descriptors: superovulation; Inbred NOD Mice; animal mating behavior; oocytes; mammalian embryo.
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spelling 2022-04-18T15:34:44Z2022-04-18T15:34:44Z2022https://repositorio.ufms.br/handle/123456789/4731The demand for new experimental models grows proportionately as pre-clinical tests need more refined and reproducible results, thus also contributing to one of the main concepts in Laboratory Animal Science: the 3 Rs, which are the reduction of animals in research, refined biomodels and techniques who seek to alleviate stress and discomfort, and replacement of experimental models with validated alternative techniques. An example of a biomodel created in the laboratory is the NOD mice (Nonobese Diabetic), which has contributed mainly to research involving type 1 diabetes mellitus, since the autoimmune process occurs in a very similar way to humans, in addition to several other areas that can be studied from this experimental model. With this in mind, the UFMS Central Animal Facility received NOD couples, which were established in the Breeding Area, where they are produced for supply to the Institution's researchers. With the establishment of this new strain, the present work sought to evaluate the reproduction of NOD mice, mated in the breeding colony, and in obtaining oocytes and embryos with and without hormonal stimulation. Data from 4 births of 20 couples kept in intensive monogamous mating system in the breeding area were used to calculate the productivity indexes. To obtain oocytes and embryos, the Whitten effect was used to synchronize estrus in all females in the study. The hormonal protocol was performed in 20 females through the administration via IP of 7.5IU of PMSG on day 0, and 48 hours later 7.5IU of hCG followed by mating of 10 females with vasectomized males (n=10) to obtain oocytes and the other 10 with intact males (n=10) to obtain embryos. On day 4, the presence of vaginal tampons and oocyte collection were observed. On day 5, embryos were collected. The same mating procedure was performed for the group of females without hormonal stimulation (n=40). In the breeding colony, a litter average of 7.50±0.68 was observed; 5.25±0.95; 2.85±0.88 and 2.20±0.73, from the first to the fourth delivery, respectively. In females that did not receive hormonal stimulation, an average of 6.25±1.15 oocytes and 2.90±0.86 embryos in the 2-cell phase (D2) were obtained, while the superovulated females obtained 79.20±8.43 oocytes and 19.90±7.90 D2 embryos. Regarding the percentage of viable embryos (D2) over the total number of embryos observed, the natural group obtained an average of 20.74±6.27 and the group of superovulated females presented 24.71±6.26. Thus, it was concluded that in intensive monogamous mating, the NOD strain shows a significant drop from the third parturition, and they present a satisfactory response to superovulation in terms of the number of oocytes and embryos, but in relation to the percentage of viable embryos there was no significant difference between groups. Therefore, superovulation is a viable option for obtaining oocytes and embryos with a reduced number of animals, being an important tool for the creation of a germplasm bank and assisted reproduction of the NOD strain. Descriptors: superovulation; Inbred NOD Mice; animal mating behavior; oocytes; mammalian embryo.A demanda por novos modelos experimentais cresce proporcionalmente à medida que os ensaios pré-clínicos necessitam de resultados mais refinados e reprodutíveis, dessa maneira contribuindo também com um dos principais conceitos na Ciência de Animais de Laboratório: os 3 Rs, sendo eles a redução do número de animais, refinamento de biomodelos e técnicas que buscam amenizar o estresse e desconforto, e a substituição (replacement) de modelos experimentais por técnicas alternativas validadas. Um exemplo de biomodelo criado em laboratório é o camundongo NOD (Nonobese Diabetic), que tem contribuído principalmente para pesquisas que envolvem o diabetes mellitus tipo 1, uma vez que o processo autoimune ocorre de maneira muito semelhante aos humanos, além de outras diversas áreas que podem ser estudadas a partir desse modelo experimental. Tendo isso em vista, o Biotério Central da UFMS recebeu casais de NOD, que foram estabelecidos na Área de Criação, onde são produzidos para fornecimento aos pesquisadores da Instituição. Com o estabelecimento dessa nova linhagem, o presente trabalho buscou avaliar a reprodução do camundongo NOD mantidos na colônia de criação e comparar a capacidade de produção de oócitos e embriões de fêmeas superovuladas e sem estímulo hormonal. Para calcular os índices de produtividade foram utilizados dados de 4 partos de 20 casais mantidos em sistema de acasalamento monogâmico intensivo na área de criação. Para obtenção de oócitos e embriões, utilizou-se o efeito Whitten para sincronização do estro em todas as fêmeas do estudo. O protocolo hormonal foi feito em 20 fêmeas através da administração via IP de 7,5UI de PMSG no dia 0, e 48 horas depois 7,5UI de hCG seguido do acasalamento de 10 fêmeas com machos vasectomizados (n=10) para obtenção de oócitos, e as outras 10 com machos inteiros (n=10) para obtenção de embriões. No dia 4 observou-se a presença de tampões vaginais e coleta dos oócitos. No dia 5 foi feita a coleta dos embriões. O mesmo procedimento de acasalamento foi feito para o grupo das fêmeas sem o estimulo hormonal (n=40). Na colônia de criação foi observada uma média da ninhada de 7,50±0,68; 5,25±0,95; 2,85±0,88 e 2,20±0,73, do primeiro ao quarto parto respectivamente. Nas fêmeas que não receberam estímulo hormonal, foram obtidos uma média de 6,25±1,15 oócitos e 2,90±0,86 embriões em fase de 2 células (D2), enquanto as superovuladas obtiveram 79,20±8,43 oócitos e 19,90±7,90 embriões D2. Em relação a porcentagem de embriões viáveis (D2) sobre o total de embriões observados, o grupo natural obteve em média 20,74±6,27 e o grupo das fêmeas superovuladas apresentou 24,71±6,26. Dessa maneira concluiu-se que em acasalamento monogâmico intensivo, a linhagem NOD apresenta uma queda significativa a partir do terceiro parto, e apresentam resposta satisfatória à superovulação quanto ao número de oócitos e embriões, mas em relação à porcentagem de embriões viáveis não houve diferença significativa entre os grupos. Portanto, a superovulação é uma opção viável para obtenção de oócitos e embriões com um número reduzido de animais, sendo uma ferramenta importante para a criação de um banco de germoplasma e reprodução assistida da linhagem NOD. Descritores: superovulação; camundongos endogâmicos NOD; comportamento sexual animal; oócitos; embrião de mamíferos.Fundação Universidade Federal de Mato Grosso do SulUFMSBrasilReprodução animalCamundongos Nonobese diabetic, BioterismoAVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisAlbert Schiaveto de SouzaAmanda Godoi Navareziinfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMSinstname:Universidade Federal de Mato Grosso do Sul (UFMS)instacron:UFMSORIGINALDissertação AMANDA GODOI NAVAREZI.pdfDissertação AMANDA GODOI NAVAREZI.pdfapplication/pdf992636https://repositorio.ufms.br/bitstream/123456789/4731/-1/Disserta%c3%a7%c3%a3o%20AMANDA%20GODOI%20NAVAREZI.pdf444ce873330d99fd6814be769409fdfaMD5-1123456789/47312022-04-18 11:34:45.975oai:repositorio.ufms.br:123456789/4731Repositório InstitucionalPUBhttps://repositorio.ufms.br/oai/requestri.prograd@ufms.bropendoar:21242022-04-18T15:34:45Repositório Institucional da UFMS - Universidade Federal de Mato Grosso do Sul (UFMS)false
dc.title.pt_BR.fl_str_mv AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)
title AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)
spellingShingle AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)
Amanda Godoi Navarezi
Reprodução animal
Camundongos Nonobese diabetic, Bioterismo
title_short AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)
title_full AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)
title_fullStr AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)
title_full_unstemmed AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)
title_sort AVALIAÇÃO REPRODUTIVA DE CAMUNDONGOS DA LINHAGEM NOD (NONOBESE DIABETIC)
author Amanda Godoi Navarezi
author_facet Amanda Godoi Navarezi
author_role author
dc.contributor.advisor1.fl_str_mv Albert Schiaveto de Souza
dc.contributor.author.fl_str_mv Amanda Godoi Navarezi
contributor_str_mv Albert Schiaveto de Souza
dc.subject.por.fl_str_mv Reprodução animal
Camundongos Nonobese diabetic, Bioterismo
topic Reprodução animal
Camundongos Nonobese diabetic, Bioterismo
description The demand for new experimental models grows proportionately as pre-clinical tests need more refined and reproducible results, thus also contributing to one of the main concepts in Laboratory Animal Science: the 3 Rs, which are the reduction of animals in research, refined biomodels and techniques who seek to alleviate stress and discomfort, and replacement of experimental models with validated alternative techniques. An example of a biomodel created in the laboratory is the NOD mice (Nonobese Diabetic), which has contributed mainly to research involving type 1 diabetes mellitus, since the autoimmune process occurs in a very similar way to humans, in addition to several other areas that can be studied from this experimental model. With this in mind, the UFMS Central Animal Facility received NOD couples, which were established in the Breeding Area, where they are produced for supply to the Institution's researchers. With the establishment of this new strain, the present work sought to evaluate the reproduction of NOD mice, mated in the breeding colony, and in obtaining oocytes and embryos with and without hormonal stimulation. Data from 4 births of 20 couples kept in intensive monogamous mating system in the breeding area were used to calculate the productivity indexes. To obtain oocytes and embryos, the Whitten effect was used to synchronize estrus in all females in the study. The hormonal protocol was performed in 20 females through the administration via IP of 7.5IU of PMSG on day 0, and 48 hours later 7.5IU of hCG followed by mating of 10 females with vasectomized males (n=10) to obtain oocytes and the other 10 with intact males (n=10) to obtain embryos. On day 4, the presence of vaginal tampons and oocyte collection were observed. On day 5, embryos were collected. The same mating procedure was performed for the group of females without hormonal stimulation (n=40). In the breeding colony, a litter average of 7.50±0.68 was observed; 5.25±0.95; 2.85±0.88 and 2.20±0.73, from the first to the fourth delivery, respectively. In females that did not receive hormonal stimulation, an average of 6.25±1.15 oocytes and 2.90±0.86 embryos in the 2-cell phase (D2) were obtained, while the superovulated females obtained 79.20±8.43 oocytes and 19.90±7.90 D2 embryos. Regarding the percentage of viable embryos (D2) over the total number of embryos observed, the natural group obtained an average of 20.74±6.27 and the group of superovulated females presented 24.71±6.26. Thus, it was concluded that in intensive monogamous mating, the NOD strain shows a significant drop from the third parturition, and they present a satisfactory response to superovulation in terms of the number of oocytes and embryos, but in relation to the percentage of viable embryos there was no significant difference between groups. Therefore, superovulation is a viable option for obtaining oocytes and embryos with a reduced number of animals, being an important tool for the creation of a germplasm bank and assisted reproduction of the NOD strain. Descriptors: superovulation; Inbred NOD Mice; animal mating behavior; oocytes; mammalian embryo.
publishDate 2022
dc.date.accessioned.fl_str_mv 2022-04-18T15:34:44Z
dc.date.available.fl_str_mv 2022-04-18T15:34:44Z
dc.date.issued.fl_str_mv 2022
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