Impact of estradiol on 3D culture of bovine oviduct epithelial cells and its effects on in vitro oviduct-embryo communication

• Grant number: 18/06674-7
• Support Opportunities: Regular Research Grants
• Duration: October 01, 2018 - September 30, 2020
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Anthony César de Souza Castilho
• Grantee: Anthony César de Souza Castilho
• Host Institution: Pró-Reitoria de Pesquisa e Pós-Graduação. Universidade do Oeste Paulista (UNOESTE). Presidente Prudente , SP, Brazil
• Associated scholarship(s): 19/02430-9 - Impact of estradiol on 3D system of bovine oviduct epithelial cell culture and its effects on in vitro communication oviduct-embryo, BP.TT

Abstract

The oviducts are known to promote a microenvironment for oocyte maturation, sperm storage and capacitation, fertilization, transport of gametes and early embryo development. Due to practical limitation of animal handling and high cost of in vivo studies, oviduct cell culture systems are a suitable and convenient model for in vitro studies, such as polarized systems (3D). The system of 3D cultivation ensures the maintenance of cell polarity, ciliary epithelium and cell viability for a longer time, proved to be a promising model for research and application. The functional control of the oviduct is multifactorial. Some studies shown transcriptional, protein and functional changes in the oviduct during the estrous cycle, antimeres ovulation site in monovulatory animals, gametes and embryo presence, number of embryos present and plasma progesterone concentration (high and low concentration, correlated to the size of the ovarian corpus luteum). In the same way, our research group has demonstrated ovarian overstimulation protocol effects on regulation of transcripts in bovine oviduct related to the contractility of the oviduct and gametes interaction. Such changes were positively correlated with high plasma concentration of estradiol in cows submitted to overstimulation protocol. Such as, production and follicular release of estradiol appear to be involved in control of cell secretion, epithelial ciliary beats frequency and smooth muscle contraction; as well as, increased estradiol is positively correlated with oviductal factors expression, the objective of this project is to investigate the transcriptional profile (RNAseq) of Bovine Oviduct Epithelial Cells (BOEC) under two conditions of cultivation: low concentrations of estradiol (Group Low, E2“) and high concentration of estradiol (Group High, E2‘; EXPERIMENT I). For this, 3D cultivation system of BOEC will be culture under two treatments with estradiol: 300 and 750 pg/mL. Besides the evaluation of the oviduct cells, the supernatant medium will be collected and processed for isolation of exossomes (EXOs), aiming to characterize the microRNA profile of EXOs (EXPERIMENT II), as well as adding these EXOs in the in vitro cultivation stage of in vitro embryo production to evaluate the embryo production efficiency and impact on the blastocyst quality (EXPERIMENT III). The reconcilement of the information provided by the transcript profile of BOEC, EXOs microRNA profile produced by the BOEC cultivation and effect of these EXOs added to in vitro embryo production under high or low concentration of estradiol conditions prospects an understanding of the direct effect and function of the estradiol in control of bovine oviduct, possibly being a strategy to impact the in vitro production of bovine embryos. (AU)

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)

FAVERO, PAOLA FRANCINI; VIEIRA DE LIMA, VICTOR AUGUSTO; DOS SANTOS, PRISCILA HELENA; MARQUES ANDRADE, ANA PAULA; MENDES, LEONARDO OLIVEIRA; PACAGNELLI, FRANCIS LOPES; DE SOUZA CASTILHO, ANTHONY CESAR. Differential fractal dimension is associated with extracellular matrix remodeling in developing bovine corpus luteum. Biochemical and Biophysical Research Communications, v. 516, n. 3, p. 888-893, AUG 27 2019. (18/06674-7, 13/11480-3)

FONTES, PATRICIA K.; RAZZA, EDUARDO M.; PUPULIM, ANTONIO G. R.; BARROS, CIRO M.; DE SOUZA CASTILHO, ANTHONY C.. Equine chorionic gonadotropin increases estradiol levels in the bovine oviduct and drives the transcription of genes related to fertilization in superstimulated cows. Molecular Reproduction and Development, v. 86, n. 11, JULY 2019. (18/06674-7, 16/25685-4)