Hostname: page-component-76fb5796d-25wd4 Total loading time: 0 Render date: 2024-04-27T11:09:55.648Z Has data issue: false hasContentIssue false
  • English
  • Français

Gene expression and protein distribution of leptin and its receptor in bovine oocytes and preattachment embryos produced in vitro

Published online by Cambridge University Press:  01 April 2009

Z. E. Madeja ,
E. Warzych ,
J. Peippo ,
D. Lechniak  and
M. Switonski
Z. E. Madeja*
Affiliation:
Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637 Poznan, Poland
E. Warzych
Affiliation:
Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637 Poznan, Poland
J. Peippo
Affiliation:
MTT Agrifood Research Finland, Biotechnology and Food Research, Jokioinen, Finland
D. Lechniak
Affiliation:
Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637 Poznan, Poland
M. Switonski
Affiliation:
Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637 Poznan, Poland
*
E-mail: zofia.madeja@bbsrc.ac.uk
Get access

Abstract

Many investigations point out the important role of leptin during the preimplantation development. Transcripts for the leptin gene (LEP) and its receptor (LEPR) have been identified in several tissues related to reproduction (e.g. ovaries, testis and oviduct) in both human and mouse. This work shows for the first time the expression and distribution patterns of LEP and LEPR in bovine oocytes and in vitro-produced embryos. Gene expression was analysed by reverse transcription PCR and real-time PCR, and the proteins were localised by immunostaining. This study included immature and mature oocytes, zygotes, two-, four-, eight- to 16-cell embryos, morulae and blastocysts and the LEP transcript was identified throughout all stages of bovine preimplantation development. However, mRNA for the LEPR gene was detected at all stages, excluding four-cell embryos. Expression of both LEP and LEPR genes was reduced at the eight- to 16-cell stage. This in addition to the absence of LEPR mRNA in four-blastomere embryos may suggest that maternally derived transcripts degenerate towards the eight- to 16-cell stage coinciding with embryonic genome activation at eight- to 16-cell stage and subsequent appearance of embryonic mRNA. Immunofluorescent staining demonstrated that LEP and LEPR proteins form a spherical rim beneath the oolemma. After maturation, however, the proteins became evenly distributed within the cytoplasm. In two- to eight-cell embryos, fluorescence was observed in the apical surface of the blastomeres, and from 10- to 16-cell stage in the apical region of outer blastomeres. This pattern persisted to the blastocyst stage, leading to LEP and LEPR distribution within trophoblast cells, but not in the inner cell mass. These results support previous findings on polar distribution of proteins within mammalian oocytes and embryos, as well as suggests leptin’s potential role during early mammalian development and implantation.

Keywords

oocyteembryoleptinleptin receptorgene expression
Type
Full Paper
Information
animal , Volume 3 , Issue 4 , April 2009 , pp. 568 - 578
Copyright
Copyright © The Animal Consortium 2008

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.)

References

Anifandis, G, Koutselini, E, Stefanidis, I, Liakopoulos, V, Leivanditis, C, Mantzavinos, T, Vamvakopoulos, N 2005. Serum and follicular fluid leptin levels are correlated with human embryo quality. Reproduction 130, 917–921.CrossRefGoogle ScholarPubMed
Antczak, M, Van Blerkom, J 1997. Oocyte influences on early development: the regulatory proteins leptin and STAT3 are polarised in mouse and human oocytes and differentially distributed within the cells of the preimplantantion stage embryo. Molecular Human Reproduction 3, 1067–1086.CrossRefGoogle ScholarPubMed
Antczak, M, Van Blerkom, J 1999. Temporal and spatial aspects of fragmentation in early human embryos: possible effects on developmental competence and association with the differential elimination of regulatory proteins from polarised domains. Human Reproduction 14, 429–447.CrossRefGoogle ScholarPubMed
Boelhauve, M, Sinowatz, F, Wolf, E, Paula-Lopes, FF 2005. Maturation of bovine oocytes in the presence of leptin improves development and reduces apoptosis of in vitro-produced blastocysts. Biology of Reproduction 73, 737–744.CrossRefGoogle ScholarPubMed
Cao, R, Brakenhielm, E, Wahlestedt, C, Thyberg, J, Cao, Y 2001. Leptin induces vascular permeability and synergistically stimulates angiogenesis with FGF-2 and VEGF. Proceedings of the National Academy of Sciences of the United States of America 98, 6390–6395.CrossRefGoogle ScholarPubMed
Castellucci, M, De Matteis, R, Meisser, A, Cancello, R, Monsurro, V, Islami, D, Sarzani, R, Marzioni, D, Cinti, S, Bischof, P 2000. Leptin modulates extracellular matrix molecules and metalloproteinases: possible implications for trophoblast invasion. Molecular Human Reproduction 6, 951–958.CrossRefGoogle ScholarPubMed
Cervero, A, Horcajadas, JA, Martin, J, Pellicer, A, Simon, CJ 2004. The leptin system during human endometrial receptivity and preimplantation development. Journal of Clinical Endocrinology and Metabolism 89, 2442–2451.CrossRefGoogle ScholarPubMed
Chomczynski, P, Sacchi, N 1987. Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry 162, 156–159.CrossRefGoogle ScholarPubMed
Cioffi, JA, Van Blerkom, J, Antczak, M, Shafer, A, Wittmer, S, Snodgrass, HR 1997. The expression of leptin and its receptors in pre-ovulatory human follicles. Molecular Human Reproduction 3, 467–472.CrossRefGoogle ScholarPubMed
Craig, J, Zhu, H, Dyce, PW, Petrik, J, Li, J 2004. Leptin enhances oocyte nuclear cytoplasmic maturation via the mitogen-activated protein kinase pathway. Endocrinology 145, 5355–5363.CrossRefGoogle ScholarPubMed
Craig, JA, Zhu, H, Dyce, PW, Wen, L, Li, J 2005. Leptin enhances porcine preimplantation embryo development in vitro. Molecular and Cellular Endocrinology 229, 141–147.CrossRefGoogle ScholarPubMed
Fedorcsak, P, Storeng, R 2003. Effects of leptin end leukemia inhibitory factor on preimplantation development and STAT3 signalling of mouse embryos in vitro. Biology of Reproduction 69, 1531–1538.CrossRefGoogle Scholar
Gonzalez, RR, Simon, C, Caballero-Campo, P, Norman, R, Chardonnens, D, Devoto, L, Bischof, P 2000. Leptin and reproduction. Human Reproduction Update 6, 290–300.CrossRefGoogle ScholarPubMed
Gurdon, JB 1992. The generation of diversity and pattern in animal development. Cell 68, 185–199.CrossRefGoogle ScholarPubMed
Holm, P, Booth, PJ, Schmidt, MH, Greve, T, Callesen, H 1999. High bovine blastocyst development in a static in vitro production system using SOFaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins. Theriogenology 52, 683–700.CrossRefGoogle ScholarPubMed
International Embryo Transfer Society (IETS) 2006. Transfers of both in vivo derived and in vitro produced embryos in cattle still on the rise and contrasted trends in other species in 2005. IETS Newsletter 24, 12–18.Google Scholar
Johnson, KM, Alvarez, X, Borkhsenious, ON, Kubisch, HM 2006. Nuclear and cytoplasmic localization of interferon-tau in in vitro-produced bovine blastocysts. Reproduction Nutrition Development 46, 97–104.CrossRefGoogle ScholarPubMed
Joudrey, EM, Lechniak, D, Petrik, J, King, WA 2003. Expression of growth hormone and its transcription factor, Pit-1, in early bovine development. Molecular Reproduction and Development 64, 275–283.CrossRefGoogle ScholarPubMed
Kane, MT, Morgan, PM, Coonan, C 1997. Peptide growth factors and preimplantation development. Human Reproduction Update 3, 137–157.CrossRefGoogle ScholarPubMed
Kawamura, K, Sato, N, Fukuda, J, Kodama, H, Kumagai, J, Takinawa, H, Nakamura, A, Tanaka, T 2002. Leptin promotes the development of mouse preimplantation embryos in vitro. Endocrinology 143, 1922–1931.CrossRefGoogle ScholarPubMed
Kawamura, K, Sato, N, Fukuda, J, Kodama, H, Kumagai, J, Takinawa, H, Murata, M, Tanaka, T 2003. The role of leptin during the development of mouse preimplantation embryos. Molecular and Cellular Endocrinology 202, 185–189.CrossRefGoogle ScholarPubMed
Kruip, TA, den Daas, JH 1997. In vitro produced and cloned embryos: effects on pregnancy, parturition and offspring. Theriogenology 47, 43–52.CrossRefGoogle Scholar
Makarevich, AV, Markkula, M 2002. Apoptosis and cell proliferation potential of bovine embryos stimulated with insulin-like growth factor I during in vitro maturation and culture. Biology of Reproduction 66, 386–392.CrossRefGoogle ScholarPubMed
Mamo, S, Baji Gal, A, Bodo, S, Dinnyes, A 2007. Quantitative evaluation and selection of reference genes in mouse oocytes and embryos cultured in vivo and in vitro. BMC Developmental Biology 7, 14, Doi: 10.1186/1471-213X-7-14.CrossRefGoogle ScholarPubMed
McDougall, K, Beecroft, J, Wasnidge, C, King, WA, Hahnel, A 1998. Sequences and expression patterns of alkaline phosphatase isozymes in preattachment bovine embryos and the adult bovine. Molecular Reproduction and Development 50, 7–17.3.0.CO;2-P>CrossRefGoogle ScholarPubMed
Paula-Lopes, FF, Boelhauve, M, Habermann, FA, Sinowatz, F, Wolf, E 2007. Leptin promotes meiotic progression and developmental capacity of bovine oocytes via cumulus cell-independent and -dependent mechanisms. Biology of Reproduction 76, 532–541.CrossRefGoogle ScholarPubMed
Robert, C, McGraw, S, Massicotte, L, Pravetoni, M, Gandolfi, F, Sirard, MA 2002. Quantification of housekeeping transcript levels during the development of bovine preimplantation embryos. Biology of Reproduction 67, 1465–1472.CrossRefGoogle ScholarPubMed
Schulz, LC, Widmaier, EP 2004. The effect of leptin on mouse trophoblast cell invasion. Biology of Reproduction 71, 1963–1967.CrossRefGoogle ScholarPubMed
Smith, JT, Waddell, BJ 2002. Leptin receptor expression in the rat placenta: changes in ob-ra, ob-rb, and ob-re with gestational age and suppression by glucocorticoids. Biology of Reproduction 67, 1204–1210.CrossRefGoogle ScholarPubMed
SPSS (Statistical Package for the Social Sciences) 2006. SPSS for windows, version 15.0. SPSS Inc., Chicago, IL, USA.Google Scholar
Stanton, JAL, Macgregor, AB, Green, DPL 2003. Gene expression in the mouse preimplantation embryo. Reproduction 125, 457–468.CrossRefGoogle ScholarPubMed
Sun, QY, Wu, GM, Lai, L, Park, KW, Cabot, R, Cheong, HT, Day, BN, Prather, RS, Schatten, H 2001. Translocation of active mitochondria during pig oocyte maturation, fertilization and early embryo development in vitro. Reproduction 122, 155–163.CrossRefGoogle ScholarPubMed
Swain, JE, Dunn, RL, McConnell, D, Gonzalez-Martinez, J, Smith, GD 2004. Direct effects of leptin on mouse reproductive function: regulation of follicular, oocyte, and embryo development. Biology of Reproduction 71, 1446–1452.CrossRefGoogle ScholarPubMed
van Tol, HTA, van Eerdenburg, FJCM, Colenbrander, B, Roelen, BAJ 2008. Enhancement of bovine oocyte maturation by leptin is accompanied by an upregulation in mRNA expression of leptin receptor isoforms in cumulus cells. Molecular Reproduction and Development 75, 578–587.CrossRefGoogle ScholarPubMed
Vigneault, C, McGraw, S, Massicotte, L, Sirard, MA 2004. Transcription factor expression patterns in bovine in vitro-derived embryos prior to maternal–zygotic transition. Biology of Reproduction 70, 1701–1709.CrossRefGoogle ScholarPubMed
Welt, CK, Schneyer, AL, Heist, K, Mantzoros, CS 2003. Leptin and soluble leptin receptor in follicular fluid. Journal of Assisted Reproduction and Genetics 20, 495–501.CrossRefGoogle ScholarPubMed
Wrenzycki, C, Herrmann, D, Lucas-Hahn, A, Korsawe, K, Lemme, E, Niemann, H 2005. Messenger RNA expression patterns in bovine embryos derived from in vitro procedures and their implications for development. Reproduction, Fertility and Development 17, 23–35.CrossRefGoogle ScholarPubMed
Yang, YJ, Cao, YJ, Bo, SM, Peng, S, Liu, WM, Duan, EK 2006. Leptin-directed embryo implantation: leptin regulates adhesion and outgrowth of mouse blastocysts and receptivity of endometrial epithelial cells. Animal Reproduction Science 92, 155–167.CrossRefGoogle ScholarPubMed
Yoon, SJ, Cha, KY, Lee, KA 2005. Leptin receptors are down-regulated in uterine implantation sites compared to interimplantation sites. Molecular and Cellular Endocrinology 232, 27–35.CrossRefGoogle ScholarPubMed
Zhao, J, Townsend, KL, Schulz, LC, Kunz, TH, Li, C, Widmaier, EP 2004. Leptin receptor expression increases in placenta, but not hypothalamus, during gestation in Mus musculus and Myotis lucifugus. Placenta 25, 712–722.CrossRefGoogle Scholar