Embriões bovinos produzidos in vitro com espermatozoides sexados por citometria de fluxo: avaliação do desenvolvimento embrionário e da expressão de genes candidatos ao reconhecimento da gestação

• Ano de defesa: 2014
• Autor(a) principal: Nonato, Amanda [UNESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Estadual Paulista (Unesp)
• Palavras-chave em Português: Bovino - Embrião; Expressão gênica; Fertilização in vitro; Citometria de fluxo; Genetica animal; Gene expression
• Link de acesso: http://hdl.handle.net/11449/110557

Doses of semen enriched with carriers of X chromosome sperm are used in artificial insemination (AI) and in embryos produced in vitro (PIVE). However, field studies using this type of semen demonstrated a reduction in pregnancy rate, which can be justified by injuries during sperm sexing and alterations in DNA, interfering with the expression of paternal genes affecting embryonic development and resulting in pregnancy loss after 90 days. The aims of this study were to compare the cleavage and blastocyst rates produced in vitro using conventional semen and sexed semen by flow cytometry, and check differences in the expression of genes related to pregnancy recognition (AKR1B1, PTGS1, PTGS2, mPGES-1, mPGES-2, TNF-α ) in embryos produced in vitro using the two types of semen. Bovine embryos were produced by in vitro fertilization, forty eight hours after fertilization the cleavage rate was evaluated, and on the seventh day after fertilization the blastocyst rate was performed. Subsequently, these blastocysts were subjected to RNA extraction for evaluation of gene expression. No significant differences (P>0.05) for cleavage and blastocyst rates between the experimental groups were observed. Related to the expression pattern, the flow cytometry did not alter the transcript levels of AKR1B1, PTGS1, PTGS2, mPGES-2 e TNF-α genes, suggesting that embryos produced with sexed semen by flow cytometry do not have detrimental changes in genes involved in pregnancy recognition. However the mPGES-1 gene was expressed 4,54 times less in embryo produced with sexed semen by flow cytometry, suggesting the possibility of pregnancy loss, since studies suggest that are required levels of expression of mPGES for maintenance and recognition of pregnancy