Author/Authors :
Sirini, Matías Angel IGEVET-Institute of Veterinary Genetic “Prof. Fernando N. Dulout” (UNLP-CONICET LA PLATA)- Faculty of Veterinary Sciences - National University of La Plata, La Plata, Buenos Aires, Argentina , Anchordoquy, Juan Patricio IGEVET-Institute of Veterinary Genetic “Prof. Fernando N. Dulout” (UNLP-CONICET LA PLATA)- Faculty of Veterinary Sciences - National University of La Plata, La Plata, Buenos Aires, Argentina , Quintana, Silvina Bee Reasearch Center - Department of Biology, FCEy N - National University of Mar del Plata - CONICET, Mar del Plata, Buenos Aires, Argentina , Furnus, Cecilia IGEVET-Institute of Veterinary Genetic “Prof. Fernando N. Dulout” (UNLP-CONICET LA PLATA)- Faculty of Veterinary Sciences - National University of La Plata, La Plata, Buenos Aires, Argentina , Relling, Alejandro Enrique IGEVET-Institute of Veterinary Genetic “Prof. Fernando N. Dulout” (UNLP-CONICET LA PLATA)- Faculty of Veterinary Sciences - National University of La Plata, La Plata, Buenos Aires, Argentina , Anchordoquy, Juan Mateo IGEVET-Institute of Veterinary Genetic “Prof. Fernando N. Dulout” (UNLP-CONICET LA PLATA)- Faculty of Veterinary Sciences - National University of La Plata, La Plata, Buenos Aires, Argentina
Abstract :
Energy balance is regulated by ghrelin which is a neuroendocrine modulator. Ghrelin is expressed in reproductive
organs. However, the role of ghrelin during in vitro maturation (IVM) and bovine preimplantational
development is limited. The purpose of this study was to measure the expression of ghrelin (GHRL) and
its receptor growth hormone secretagogue receptor 1A (GHS-R1A) mRNA, and determine cumulus oocyte
complex (COC) viability after IVM with 0, 20, 40 and 60 pM of ghrelin. Also, pronuclear formation was
recorded after in vitro fertilization (IVF). GHRL and GHS-R1A mRNA expression in oocyte and cumulus
cells (CCs) was assessed using reverse transcription-polymerase chain reaction (PCR). Oocyte and
CC viability were analyzed with the fluorescein diacetate fluorochrome-trypan blue technique. Pronuclear
formation was determined 18 hours after IVF with Hoechst 33342. The results demonstrated that ghrelin
mRNA is present in oocyte and CCs before and after 24 hours IVM with all treatments. Ghrelin receptor,
GHS-R1A, was only detected in oocytes and CCs after 24 hours IVM with 20, 40 and 60 pM of ghrelin.
Oocyte viability was not significantly different (P=0.77) among treatments. However, CC viability was
significantly lower (P=0.04) when COCs were matured with ghrelin (77.65, 72.10, 66.32 and 46.86% for
0, 20, 40, and 60 pM of ghrelin, respectively). The chance of two pronuclei forming were higher (P=0.03)
when ghrelin was not be added to the IVM medium. We found that ghrelin negatively impacts CC viability
and pronuclear formation.
Keywords :
mRNA Expression , Ghrelin , GHS-R1A , In Vitro Oocyte Maturation
