title: Daily supplementation with ghrelin improves in vitro bovine blastocysts formation rate and alters gene expression related to embryo quality.
abstract: Ghrelin is a gastric peptide having regulatory role in the reproductive system functionality, acting mainly at central level. Because the expression of ghrelin system (ghrelin and its receptor) has been detected in the bovine ovary, the objectives of the present study were to investigate whether ghrelin can affect the developmental potential of in vitro-produced embryos, and to test their quality in terms of relative abundance of various genes related to metabolism, apoptosis and oxidation. In the first experiment, in vitro-produced zygotes were cultured in the absence (control [C]) and in the presence of three concentrations of acylated ghrelin (200 pg/mL [Ghr200], 800 pg/mL [Ghr800]; and 2000 pg/mL [Ghr2000]); blastocyst formation rates were examined on Days 7, 8, and 9. In the second experiment, only the 800 pg/mL dose of ghrelin was used. Zygotes were produced as in experiment 1 and 24 hours post insemination they were divided into 4 groups; in two groups (C; without ghrelin; Ghr800 with ghrelin), embryos were cultured without medium replacement; in the remaining two groups (Control N and GhrN), the culture medium was daily renewed. A pool of Day-7 blastocysts were snap frozen for relative mRNA abundance of various genes related to metabolism, oxidation, implantation, and apoptosis. In experiment 3, embryos were produced as in experiment 2, but in the absence of serum (semi-defined culture medium). In experiment 1, no differences were detected between C, Ghr200, and Ghr2000, although fewer blastocysts were produced in group Ghr800 compared with C. In experiment 2, the lowest blastocysts yield was found in Ghr800, whereas daily renewal of ghrelin (Ghr800N) resulted to increased blastocysts formation rate, which on Day 7 was the highest among groups (P < 0.05). In experiment 3, ghrelin significantly suppressed blastocysts yield. Significant differences were detected in various relative mRNA abundance, giving an overall final notion that embryos produced in the presence of ghrelin were of better quality than controls. Our results imply a specific role of ghrelin in early embryonic development; however, the specific mode of its action needs further investigation.
authors: Department of Obstetrics & Reproduction, Veterinary Faculty, University of Thessaly, Karditsa, Greece; Department of Obstetrics & Gynecology, Faculty of Medicine, University of Thessaly, Larissa, Greece.
authors: Eleni
authors: E
authors: Dovolou
authors: Departmento de Reproducción Animals y Conservación de Recursos Zoogeneticos, INIA, Madrid, Spain.
authors: Eva
authors: E
authors: Periquesta
authors: Department of Obstetrics & Gynecology, Faculty of Medicine, University of Thessaly, Larissa, Greece.
authors: Ioannis E
authors: IE
authors: Messinis
authors: Veterinary Research Institute, Ionia Thessaloniki, Greece.
authors: Theodora
authors: T
authors: Tsiligianni
authors: Department of Obstetrics & Gynecology, Faculty of Medicine, University of Thessaly, Larissa, Greece.
authors: Konstantinos
authors: K
authors: Dafopoulos
authors: Departmento de Reproducción Animals y Conservación de Recursos Zoogeneticos, INIA, Madrid, Spain.
authors: Alfonso
authors: A
authors: Gutierrez-Adan
authors: Department of Obstetrics & Reproduction, Veterinary Faculty, University of Thessaly, Karditsa, Greece. Electronic address: gsamir@vet.uth.gr.
authors: Georgios S
authors: GS
authors: Amiridis
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D002417:Cattle / Q000235:genetics / Q000378:metabolism*; D047108:Embryonic Development / Q000235:genetics / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D054439:Ghrelin / Q000378:metabolism*; D012313:RNA / Q000737:chemistry / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary
chemical_list: D054439:Ghrelin; D012313:RNA
keywords: Bovine blastocysts; DNMT3A; Ghrelin; IGF2R; In vitro culture