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Role of inflammatory pathways on embryonic competence and cryotolerance

Grant number: 19/18415-9
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): December 01, 2019
Effective date (End): July 31, 2023
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal researcher:Mateus José Sudano
Grantee:Roniele Santana Valente
Home Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil

Abstract

One of the main challanges to further expansion of in vitro produced of embryos is its reduced cryosurvival. When exposed to stressful situations, cells develop self-defense mechanisms that, under exacerbated conditions, can lead to cell death. NF-KB (nuclear factor kappa B) is an inducible transcription factor known to be a regulator of inflammatory cellular response and the development of defense mechanisms against a wide variety of stimuli, controlling the expression of numerous target genes. In order to elucidate the participation of NF-kB in embryonic survival mechanisms, bovine embryos will be produced in vitro with the supplementation of NF-kB modulating agents, TNF (tumor necrosis factor) inducer, IL-13 (Interleukin 13) inhibitor, the association of both (IL-13 + TNF), and subsequently compared to a control group. Embryonic development rates, relative abundance of inflammatory genes transcripts and oxidative stress will be evaluated. Blastocysts will be evaluated for cell viability (Tunel) and protein expression of NF-Kb, IKB±, CASP3 and 8-OxoG by imonofluorescence.The culture medium will be quantified for the presence of inflammatory interleukins. Secondly, the same variables will be evaluated in embryos treated with IL-13 and control subjected to stress conditions (cold and presence of H2O2) during culture. Finally, the cryotolerance of vitrified control and IL-13 treated embryos will be evaluated in two different situations: in the embryonic development medium and in the reculture medium after warming. For statistical analysis, univariate data models will be used, assuming a significance level of 5%. (AU)