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Efeito do antioxidante Alfa-Tocoferol na taxa de maturação de ovócitos bovinos

Authors: Valadão, Lóide Isabel Simões Soares;

Efeito do antioxidante Alfa-Tocoferol na taxa de maturação de ovócitos bovinos

Abstract

A produção in vitro de embriões é uma técnica com fins experimentais que é também utilizada como ferramenta de campo, para o melhoramento reprodutivo e genético das explorações. O presente trabalho foi delineado para avaliar o efeito do antioxidante Alfa-Tocoferol na taxa de maturação de ovócitos bovinos. Para tal, foram recolhidos no Matadouro do IAMA da Ilha Terceira, um total de 194 ovários transportados para o laboratório em solução salina, sendo processados no máximo 2 horas após o abate. Foram feitas no total 12 manipulações. Os folículos de 2 a 8mm de diâmetro, foram puncionados tendo sido conseguidos um total de 779 complexos de cumulus – ovocitários (COCs) considerados de qualidade 1 e 2 de acordo com o seu aspeto morfológico. Estes COCs foram divididos em 4 grupos, tendo sido cultivados em meio de maturação suplementado com diferentes concentrações (0; 0,5; 1 e 2mM) de Alfa-Tocoferol durante 24h a 38,5°C, com 5% de CO₂ no ar e saturada de humidade. Após este período, os COCs foram transferidos para tubos cónicos de 15mL devidamente identificados que continham 2-3mL de meio de lavagem. Posteriormente, as células do cumulus foram removidas, os ovócitos foram corados com aceto-orceína e observados ao microscópio de contraste de fase, procedendo-se à avaliação das diferentes fases nucleares nas diferentes concentrações de α-Tocoferol. Os resultados da maturação in vitro foram expressos em percentagem. O antioxidante α-Tocoferol numa concentração de 0,5mM fez aumentar significativamente (P<0.05) a taxa de maturação, relativamente ao grupo de controlo, tendo os resultados sido, respetivamente de 60,87 vs 68,00. Para as outras duas concentrações, não foram observadas diferenças significativas quando comparados com os resultados obtidos no grupo de controlo, tendo a taxa de maturação sido respetivamente de 64,29 e de 60,61, respetivamente para as concentrações de 1 e 2mM de α-Tocoferol. Este estudo sugere-nos que adição do antioxidante α-Tocoferol na concentração de 0,5mM aumenta a Taxa de Maturação dos ovócitos da espécie bovina. Futuramente, seria importante complementar este estudo com a realização de trabalhos sobre o efeito deste antioxidante sobre a taxa de produção in vitro de embriões, bem como a suplementação da alimentação das fêmeas bovinas com Vitamina E.

ABSTRACT: The in vitro production of embryos is a technique with experimental purposes that is also used as a field tool for the breeding and genetic improvement of the farms. The present work was designed to evaluate the effect of the antioxidant Alpha-Tocopherol on the maturation rate of bovine oocytes. For this purpose, a total of 194 ovaries transported to the laboratory in saline were collected at the IAMA Slaughterhouse in Terceira Island, being processed no more than 2 hours after slaughter. A total of 12 manipulations were made. The follicles 2 to 8 mm in diameter were punctured and a total of 779 cumulus - oocyte complexes (COCs) were obtained, considered of quality 1 and 2 according to their morphological aspect. These COCs were divided into 4 groups and grown in maturation medium supplemented with different concentrations (0, 0.5, 1 and 2 mM) of alpha-Tocopherol for 24 h at 38.5°C, with 5% CO₂ in the air and saturated with moisture. After this period, the COCs were transferred to properly identified 15mL conical tubes containing 2-3mL of wash medium. Subsequently, the cumulus cells were removed, the oocytes were stained with aceto-orcein and observed under the phase contrast microscope, and the different nuclear phases were evaluated at the different α-Tocopherol concentrations. In vitro maturation results were expressed as a percentage. The α-Tocopherol antioxidant at a concentration of 0.5mM increased significantly (P <0.05) the maturation rate, relative to the control group, with the results being respectively 60.87 vs 68.00. For the other two concentrations, no significant differences were observed when compared to the results obtained in the control group, the maturation rate being respectively 64.29 and 60.61, respectively for the concentrations of 1 and 2mM α-Tocopherol. This study suggests that addition of the antioxidant α-Tocopherol at the concentration of 0.5mM increases the Maturation Rate of oocytes of the bovine species. I the future, it would be important to complement this study with the work carried out on the effect of this antioxidant on the rate of in vitro production of embryos, as well as the supplementation of the feeding of the bovine females with Vitamin E.

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Portugal
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Keywords

Fecundação in vitro, Meiose, Antioxidantes, Stress Oxidativo, Bovinos

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selected citations
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This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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