Hostname: page-component-78c5997874-mlc7c Total loading time: 0 Render date: 2024-11-10T14:43:16.411Z Has data issue: false hasContentIssue false

Effects of ghrelin on activation of Akt1 and ERK1/2 pathways during in vitro maturation of bovine oocytes

Published online by Cambridge University Press:  07 March 2017

Thomas-Markos Chouzouris
Affiliation:
Department of Obstetrics and Reproduction, Veterinary Faculty, School of Health Sciences, Karditsa, University of Thessaly, Greece.
Eleni Dovolou
Affiliation:
Department of Obstetrics and Reproduction, Veterinary Faculty, School of Health Sciences, Karditsa, University of Thessaly, Greece.
Fotini Krania
Affiliation:
Department of Obstetrics and Reproduction, Veterinary Faculty, School of Health Sciences, Karditsa, University of Thessaly, Greece.
Ioannis S. Pappas
Affiliation:
Department of Pharmacology, Veterinary Faculty, School of Health Sciences, Karditsa, University of Thessaly, Greece.
Konstantinos Dafopoulos
Affiliation:
Department of Obstetrics and Gynecology, Faculty of Medicine, School of Health Sciences, Larissa, University of Thessaly, Greece.
Ioannis E. Messinis
Affiliation:
Department of Obstetrics and Gynecology, Faculty of Medicine, School of Health Sciences, Larissa, University of Thessaly, Greece.
George Anifandis
Affiliation:
Department of Obstetrics and Gynecology, Faculty of Medicine, School of Health Sciences, Larissa, University of Thessaly, Greece.
Georgios S. Amiridis*
Affiliation:
PO Box 199, 43100 Karditsa, Greece. Department of Obstetrics and Reproduction, Veterinary Faculty, School of Health Sciences, Karditsa, University of Thessaly, Greece.
*
All correspondence to: Georgios S. Amiridis. PO Box 199, 43100 Karditsa, Greece. Tel: +302441066073. E-mail gsamir@vet.uth.gr

Summary

The purpose of this study was to investigate the possible molecular pathways through which ghrelin accelerates in vitro oocyte maturation. Bovine cumulus–oocyte complexes (COCs), after 18 or 24 h maturation in the absence or the presence of 800 pg ml–1 of acylated ghrelin were either assessed for nuclear maturation or underwent in vitro fertilization in standard media and putative zygotes were cultured in vitro for 8 days. In a subset of COCs the levels of phosphorylated Akt1 and ERK1/2 (MAPK1/3) were assessed at the 0th, 6th, 10th, 18th and 24th hours of in vitro maturation (IVM). At 18 and 24 h no difference existed in the proportion of matured oocytes in the ghrelin-treated group, while in the control group more (P < 0.05) matured oocyte were found at 24 h. Oocyte maturation for 24 h in the presence of ghrelin resulted in substantially reduced (P < 0.05) blastocyst yield(16.3%) in comparison with that obtained after 18 h (30.0%) or to both control groups (29.3% and 26.9%, for 18 and 24 h in maturation, respectively). Ghrelin-treated oocytes expressed lower Akt1 phosphorylation rate at the 10th hour of IVM, and higher ERK1/2 at the 6th and 10th hours of IVM compared with controls. In cumulus cells, at the 18th and 24th hours of IVM Akt1 phosphorylation rate was higher in ghrelin-treated oocytes. Our results imply that ghrelin acts in a different time-dependent manner on bovine oocytes and cumulus cells modulating Akt1 and ERK1/2 phosphorylation, which brings about acceleration of the oocyte maturation process.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2017 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

5

These authors contributed equally to this work and are listed alphabetically.

References

Bai, R., Zhao, P., Cao, G., Wen, S., Li, Q. & Meng, Q. (2012). Ghrelin promotion of oocyte maturation via ERK1/2 pathway in ovis aries. Cell. Mol. Biol. 58 (Suppl), 1797–802.Google ScholarPubMed
Chouzouris, T.M., Dovolou, E., Dafopoulos, K., Georgoulias, P., Vasileiou, N.G., Fthenakis, G.C., Anifandis, G. & Amiridis, G.S. (2016). Ghrelin suppresses the GnRH-induced preovulatory gonadotropin surge in dairy heifers. Theriogenology 86, 1615–21.CrossRefGoogle Scholar
Chung, H., Seo, S., Moon, M. & Park, S. (2008). Phosphatidylinositol-3-kinase/Akt/glyucogen synthetase kinase-3β and ERK 1/2 pathways mediated protective effects of acylated and unacylated ghrelin against oxygen-glucose deprivation-induced apoptosis in primary cortical neuronal cells. J. Endocrinol. 198, 511–21.CrossRefGoogle Scholar
Delhanty, P.J.D., van der Eerden, B.C., van der Velde, M., Gauna, C., Pols, H.A., Jahr, H., Chiba, H., van der Lely, A.J. & van Leeuwen, J.P.T.M. (2006). Ghrelin and unacylated ghrelin stimulate human osteoblast growth via mitogen activated protein kinase (MAPK)/phosphoinositide 3-kinase (PI3K) pathways in the absence of GHS-R1a. J. Endocrinol. 188, 3747.Google Scholar
Dovolou, E., Chadio, S., Messinis, I.E., Rekkas, C.A., Deligiannis, C., Kalogiannis, D. & Amiridis, G.S. (2013). Human ghrelin decreases pituitary response to GnRH in superovulated ewes. Theriogenology 80, 262–8.Google Scholar
Dovolou, E., Clemente, M., Amiridis, G.S., Messinis, I., Kallitsaris, A., Gutierrez-Adan, A. & Rizos, D (2011). Effects of guaiazulene on in vitro bovine embryo production and on mRNA transcripts related to embryo quality. Reprod. Dom. Anim. 46, 862–9.Google Scholar
Dovolou, E., Messinis, I.E., Periquesta, E., Dafopoulos, K., Gutierrez-Adan, A. & Amiridis, G.S. (2014a). Ghrelin accelerates in vitro maturation of bovine oocytes. Reprod. Domest. Anim. 49, 665–72.Google Scholar
Dovolou, E., Periquestam, E., Messinism, I.E., Tsiligianni, Th., Dafopoulos, K., Gutierrez-Adan, A. & Amiridis, G.S. (2014b). Daily supplementation with ghrelin improves in vitro bovine blastocysts formation rate and alters gene expression related to embryo quality. Theriogenology 81, 565–71.CrossRefGoogle ScholarPubMed
Fissore, R.A., He, C.L. & Vande Woude, G.F. (1996). Potential role of mitogen-activated protein kinase during meiosis resumption in bovine oocytes. Biol. Reprod. 55, 1261–70.Google Scholar
Granata, R., Settanni, F., Biancone, L., Trovato, L., Nano, R., Bertuzzi, F., Destefanis, S., Annunziata, M., Martinetti, M., Catapano, F., Che, C., Isgaard, J., Papotti, M., Ghigo, E. & Muccioli, G. (2007). Acylated and unacylated ghrelin promote proliferation and inhibit apoptosis of pancreatic β–cells and human islets: involvement of 3′,5′ cyclic adenosine monophosphate/protein kinase A, extracellular signal–regulated kinase ½ and phosphatidyl inositol 3-kinase/akt signaling. Endocrinology 148, 512–29.CrossRefGoogle Scholar
Kim, M.S., Yoon, C.Y., Jang, P.G., Park, Y.J., Shin, C.S., Park, H.S., Ryu, J.W., Park, Y.K., Park, J.Y., Lee, K.U., Kim, S.Y., Lee, H.K., Kim, Y.B. & Park, K.S. (2004). The mitogenic and antiapoptotic actions of ghrelin in 3T3-L1 adipocytes. Mol. Endocrinol. 18, 2291–301.Google Scholar
Li, L., Ferin, M., Sauer, M.V. & Lobo, R.A. (2011). Serum and follicular fluid ghrelin levels negatively reflect human oocyte quality and in vitro embryo development. Fertil. Steril. 96, 116–20.CrossRefGoogle ScholarPubMed
Nuttinck, F., Reinaud, P., Tricoire, H., Vigneron, C., Peynot, N., Mialot, J.P., Mermillod, P. & Charpigny, G. (2002). Cyclooxygenase-2 is expressed by cumulus cells during oocyte maturation in cattle. Mol. Reprod. Dev. 61, 93101.CrossRefGoogle ScholarPubMed
Popelkova, M., Sirotkin, A.V., Bezakova, A., Makarevic, A.V., Pivko, J., Kacmarik, J. & Kovac, G. (2006). Effect of IGF-I, leptin, ghrelin and MAPK-ERK on the nuclear maturation of bovine oocytes. Bull. Vet. Inst. Pulawy. 50, 179–81.Google Scholar
Salhab, M., Dhorne-Pollet, S., Auclair, S., Guyader-Joly, C., Brisard, D., Dalbies-Tran, R., Dupont, J., Ponsart, C., Mermillod, P. & Uzbekova, S. (2013). In vitro maturation of oocytes alters gene expression and signaling pathways in bovine cumulus cells. Mol. Reprod. Dev. 80, 166–82.Google Scholar
Salhab, M., Tosca, L., Cabau, C., Papillier, P., Perreau, C., Dupont, J., Mermillod, P. & Uzbekova, S. (2011). Kinetics of gene expression and signaling in bovine cumulus cells throughout IVM in different mediums in relation to oocyte developmental competence., cumulus apoptosis and progesterone secretion. Theriogenology 75, 90104.Google Scholar
Sirard, M.A., Florman, H.M., Leibfried-Rutledge, M.L., Barnes, F.L., Sims, M.L. & First, N.L. (1989). Timing of nuclear progression and protein synthesis necessary for meiotic maturation of bovine oocytes. Biol. Reprod. 40, 1257–63.CrossRefGoogle ScholarPubMed
Sutton, M.L., Cetica, P.D., Beconi, M.T., Kind, K.L., Gilchrist, R.B. & Thompson, J.G. (2003). Influence of oocyte-secreted factors and culture duration on the metabolic activity of bovine cumulus cell complexes. Reproduction 126, 2734.Google Scholar
Suzuki, H., Sasaki, Y., Shimizu, M., Matsuzaki, M., Hashizume, T. & Kuwayama, H. (2010) Ghrelin and leptin did not improve meiotic maturation of poscine oocytes cultured in vitro . Reprod. Dom. Anim. 45, 927–30.Google Scholar
Tomek, W. & Smiljakovic, T. (2005). Activation of Akt (protein kinase B) stimulates metaphase I to metaphase II transition in bovine oocytes. Reproduction 130, 423–30.CrossRefGoogle ScholarPubMed
Tosca, L., Uzbekova, S., Chabrolle, C. & Dupont, J. (2007). Possible role of 5′AMP-activated protein kinase in the metformin-mediated arrest of bovine oocytes at the germinal vesicle stage during in vitro maturation. Biol. Reprod. 77, 452–65.Google Scholar
Torner, H., Kubelka, M., Heleil, B., Tomek, W., Aim, H., Kuzmina, T. & Guiard, V. (2001). Dynamics of meiosis and protein kinase activities in bovine oocytes correlated to prolactin treatment and follicle size. Theriogenology 55, 885–99.Google Scholar
Veldhuis, J.D. & Bowers, C.Y. (2010). Integrating GHS into the ghrelin system. Int. J. Pept. Article ID 879503., 40 pp., https://doi.org/10.1155/2010/879503.CrossRefGoogle Scholar
Verlhac, M.H., Kubiak, J.Z., Clarke, H.J. & Maro, B. (1994). Microtubule and chromatin behavior follow MAP kinase activity but not MPF activity during meiosis in mouse oocytes. Development 120, 1017–25.Google Scholar
Vigneron, C., Perreau, C., Dupont, J., Uzbekova, S., Prigent, C. & Mermillod, P. (2004). Several signaling pathways are involved in the control of cattle oocyte maturation. Mol. Reprod. Dev. 69, 466–74.Google Scholar