In vitro culture of bovine steroidogenic and non-steroidogenic luteal cells: effect of PGF2a on the expression of genes and proteins associated with steroidogenesis and apoptosis

• Grant number: 15/01940-2
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: April 01, 2015
• End date: September 30, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: João Carlos Pinheiro Ferreira
• Grantee: Flavia Caroline Destro
• Supervisor: Joy Lee Pate
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Institution abroad: Pennsylvania State University, United States
• Associated to the scholarship: 13/00992-3 - In vitro culture of bovine steroidogenic luteal cells and non-steroidogenic. Effect of PGF2a on the expression of genes and proteins associated with steroidogenesis and apoptosis, BP.DR

Abstract

This work aims to perform in vitro culture of bovine luteal cells evaluate the effect of PGF2± on gene expression and protein in substances involved in steroidogenesis (StAR), the production of PGF2± intraluteal (PGTS, PTGFS, HPGD) in the induction of apoptosis (CASP-3, Bax, Bcl2) and genes of immune system that have an increased expression during the process of luteolysis and functional intraluteal that stimulate the synthesis of prostaglandin (IL-8 and FAS) in culture of cells in the corpus luteum. To do so will be conducted one experiment. The CLs will be obtained from local slaughterhouses, processed and will be realized co-culture of steroidogenic luteal cells (CLEs), endothelial luteal cells (CLEn) and immune system cells (CSI) in order to observe the action of PGF2± in these cell types in vitro. The CLEs will be cultured again for 24 hours and then CLEn and CSI will be added and pre-incubated for 2 hours (co-culture) and the same treatments and evaluations. This cells will then receive the following treatments: Control Group: 0.5mL of culture medium, which will be partially substituted after 24 hours of culture for 0.5mL culture medium and then consecutively from 6 to 6 hours; PGF2 ± Group: 10-6M PGF2± (Sigma® P 0424; KORZEKWA et al., 2008) diluted in 0.5mL culture medium. After 24 hours, this medium will be partially substituted by 10-6M PGF2± diluted in 0.5mL culture medium from 6 to 6 hours until the completion of 18 hours. Cultured cells will be evaluated by using gene expression (qRT-PCR) for STAR, PTGS2, PTGFS, HPGD, IL-8, FAS, CASP-3, Bax, Bcl-2 and finally will be dosed the progesterone (P4) of the medium culture by radioimmunoassay (RIA). Data will be recorded and adequate statistical tests will be performed. (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)

DESTRO, F. C.; MARTIN, I.; LANDIM-ALVARENGA, F. D. C.; FERREIRA, J. C. P.; PATE, J. L.. Effects of concanavalin A on the progesterone production by bovine steroidogenic luteal cells in vitro. REPRODUCTION IN DOMESTIC ANIMALS, v. 51, n. 5, p. 848-852, OCT 2016. (13/07439-8, 15/01940-2, 13/00992-3)