Influence of EGF on the viability and activation of primordial follicles in ovarian tissue from prepubertal versus adults bitches

• Grant number: 15/17412-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: January 11, 2016
• End date: July 10, 2016
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Maria Denise Lopes
• Grantee: Leda Maria Costa Pereira
• Supervisor: Nucharin Songsasen
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Institution abroad: Smithsonian Conservation Biology Institute (SCBI), United States
• Associated to the scholarship: 13/21667-3 - IDENTIFICATION OF MARKERS OF CELL CYCLE COMPETENCE INVOLVED IN THE CANINE OOCYTE PROTEOMIC ANALYSIS, BP.DR

Abstract

This study will evaluate the influences of epidermal growth factor (EGF) on in vitro viability and activation of primordial follicles from ovarian tissue of prepubertal (age, <6 months) versus adult (age, >8 months) female dogs. The EGF dose ranges will be selected based on studies that used ovarian tissue culture in other species and two experiments will be conducted. Experiment one will evaluate downstream pathways that have influence primordial follicle viability. For that, varian cortical pieces from five prepubertal and four adult bitches will be allocated randomly into eight treatment groups: 1] no factor (control), 2] 100 ng/ml of EGF, 3] 100 ng/ml of EGF plus 0.1 µM AG1478 (EGFR inhibitor), 4] 100 ng/ml of EGF plus 1 µM AG1478, 5] 100 ng/ml of EGF plus 1µM U0126 (MAPK inhibitor), 6] 100 ng/ml of EGF plus 10 µM U0126, 7] 100 ng/ml of EGF plus 1µM LY294002 (PI3K inhibitor), or 8] 100 ng/ml of EGF plus 10 µM LY294002. Fresh ovarian tissue and tissue cultured for 14 days will be evaluated for viability and morphology. After extracting proteins from fresh and cultured tissue, we will be examine phosphorylation of MAPK and AKT by Western blot analysis. Experiment 2 will be evaluate the influence of the EGF signaling pathway on ovarian cell proliferation. Cell proliferation in fresh versus cultured tissue will be evaluated by PCNA and EdU staining

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)

COSTA PEREIRA, LEDA MARIA; THONGKITTIDILOK, CHOMMANART; LOPES, MARIA DENISE; SONGSASEN, NUCHARIN. Effect of anti-apoptotic drug Z-VAD-FMK on in vitro viability of dog follicles. Theriogenology, v. 122, p. 124-129, DEC 2018. (14/19776-1, 13/21667-3, 15/17412-5)