CEDIE   05498
CENTRO DE INVESTIGACIONES ENDOCRINOLOGICAS "DR. CESAR BERGADA"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
CCR2 Receptor and Its Chemokine Ligands in the Feline Cumulus Oocyte Complex (COC) and Antral Follicle Wall
Autor/es:
JAWORSKI, JP; ROJO, JL; PELUFFO, MC
Lugar:
Washington DC
Reunión:
Congreso; 50th SSR Annual Meeting; 2017
Institución organizadora:
Society for the Study of Reproduction (SSR)
Resumen:
The aim of this study was to: 1) evaluate mRNA expression of CCR2, its ligands (MCP1, MCP2, MCP3 and MCP4) and genes related with periovulatory events (AREG and HAS2) within the cumulus oocyte complex (COC) and follicle wall after LH stimulus, using feline antral follicle culture; 2) evaluate mRNA expression of CCR2, MCP1, AREG and HAS2 within the COC, using feline COC culture in the presence of MCP1. Both culture systems were developed by our laboratory and exhibit physiological response to gonadotropin stimulus. First, ovaries were removed from adult female domestic cats (Felis catus, n=52) during the breeding season at unknown stages of the estrous cycle. Antral follicles larger than 0.5 mm were mechanically dissected from the ovary, measured and individually cultured (n=130) for 6, 12, 24 or 36 hr in the absence or presence of recombinant human LH (75 mIU/ml). At the end of the culture period, COCs and follicle walls were dissected and used for RNA extraction. Following cDNA synthesis, mRNA expression was assessed by quantitative real-time PCR (qPCR). Also, a subset of COCs (n=20) were fixed for CCR2 immunofluorescence. In a second experiment, COCs (n=28) were isolated from antral follicles larger than 0.5 mm (n=7 animals) and cultured for 3hr (time when periovulatory genes peak in our culture system) in the presence or absence of recombinant MCP1 (10 and 100 ng/ml). COCs were fixed or stored for RNA extraction and qPCR. Two-way ANOVA analyses (Time-Treatment) showed significant effects (p