Role of cholesterol in criotolerance and viability of in vitro produced bovine embryos with different development kinetics.

• Grant number: 16/05986-0
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2016
• End date: September 30, 2019
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Marcella Pecora Milazzotto
• Grantee: Kelly Annes
• Host Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil
• Associated scholarship(s): 18/01965-3 - The role of cholesterol in cryobiology, apoptosis and lipids content of in vitro produced (IVP) bovine embryos, BE.EP.DR

Abstract

Embryos with high amounts of lipids have their impaired development, with increased embryo death and lower survival after cryopreservation. Additionally, the lipid content is associated with lower pregnancy rates and low success cryopreservation of IVP embryos when compared to embryos produced in vivo. A component which is present in both lipid droplets as the membranes is cholesterol, which has been suggested by recent studies as a major lipid to be investigated more closely, having demonstrated differences in the amounts of related genes transcribed this pathway in embryos cultured in the presence and absence of serum, as well as embryos with different kinetics as compared to those produced in vivo. Therefore, the aim of this study is to investigate the role of cholesterol in embryos of different development kinetics, as well as their influence on the cryopreservation and other variables of embryo viability. For this, bovine embryos are produced in vitro in culture medium supplemented with inhibitory drugs (simvastatin) and of stimulatory (IGF-1) biosynthesis of cholesterol. The embryos are classified as development kinetics of the 40hpi collected at blastocyst stage and evaluated: the amount of lipid droplets (Nile Red), cholesterol profile (mass spectrometry), the quantitation of transcripts related with cholesterol metabolic pathway (RT-PCR), the induction of apoptosis (caspase / TUNEL), immunolocalization of important proteins in the cholesterol metabolic pathway (Western Blot) and cryopreservation.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ANNES, KELLY; MUELLER, DIEGO B.; VILELA, JORGE A. P.; VALENTE, RONIELE S.; CAETANO, DIANA P.; CIBIN, FRANCIELLI W. S.; MILAZZOTTO, MARCELLA P.; MESQUITA, FERNANDO S.; BELAZ, KATIA R. A.; EBERLIN, MARCOS N.; et al. Influence of follicle size on bovine oocyte lipid composition, follicular metabolic and stress markers, embryo development and blastocyst lipid content. REPRODUCTION FERTILITY AND DEVELOPMENT, v. 31, n. 3, p. 462-472, FEB 2019. (16/05986-0)

ANNES, KELLY; DE LIMA, CAMILA BRUNA; ISPADA, JESSICA; DOS SANTOS, ERIKA CRISTINA; FONTES, PATRICIA KUBO; NICHI, MARCILIO; GOUVEIA NOGUEIRA, MARCELO FABIO; SUDANO, MATEUS JOSE; MILAZZOTTO, MARCELLA PECORA. Insulin-like growth factor-1 (IGF-1) selectively modulates the metabolic and lipid profile of bovine embryos according to their kinetics of development. Theriogenology, v. 204, p. 7-pg., 2023-04-06. (17/18384-0, 16/05986-0)