Volume 11, Issue 9 (12-2013)                   IJRM 2013, 11(9): 733-0 | Back to browse issues page

XML Persian Abstract Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Rajabpour-Niknam M, Totonchi M, Shahhosseini M, Farrokhi A, Alipour H, Eftekhari-Yazdi P. Quantitative expression of developmental genes, Pou5f1 (Oct4) and Mest (Peg1), in vitrified mouse embryos. IJRM 2013; 11 (9) :733-0
URL: http://ijrm.ir/article-1-460-en.html
1- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
2- Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
3- Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
4- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran , peyazdi@yahoo.com
Abstract:   (2502 Views)
Background: Embryo cryopreservation is the process that water is removed from the cell by cryoprotectant materials, and embryos are stored at temperature below zero. This process may affect the viability and developmental potential of embryos.
Objective: In this study, the effect of the vitrification cryotop method on the expression level of Oct4 and Mest developmental genes in mouse blastocysts was examined.
Materials and Methods: The collected 2-cell embryos of superovulated mouse by oviduct flushing were divided into non-vitrified and vitrified groups. These embryos were cultured to the blastocyst stage directly in the non-vitrified group and in the vitrified group, these embryos were cultured to 4-8 cell embryos, vitrified with cryotop in these stages and after 2-6 months, warmed and cultured to blastocyst embryos. Quantitative expression of two developmental genes, namely Oct4 and Mest, were performed in these groups, using RNA purification and Real-time RT-PCR.
Results: Quantitative PCR analysis showed that the expression level of both genes, Oct4 and Mest, was reduced significantly in the vitrified-warmed group relative to the control group (p=0.046 and p=0.001).
Conclusion: This study revealed that morphologically normal embryos show a reduced amount of Oct4 and Mest transcripts which indicate that the vitrification method negatively effects the expression level of these two developmental genes.
Full-Text [PDF 126 kb]   (691 Downloads) |   |   Full-Text (HTML)  (346 Views)  
Type of Study: Original Article |

1. Ling XF, Zhang JQ, Cao SR, Chen J, Peng Y, Guo X, et al. Effect of cryotop vitrification on preimplantation developmental competence of murine morula and blastocyst stage embryos. Reprod Biomed Online 2009; 19: 708-713. [DOI:10.1016/j.rbmo.2009.09.006]
2. Kuwayama M. Highly efficient vitrification for cryopreservation of human oocytes and embryos: the Cryotop method. Theriogenology 2007; 67: 73-80. [DOI:10.1016/j.theriogenology.2006.09.014]
3. Wikland M, Hardarson T, Hillensjӧ T, Westin C, Westlander G, Wood M, et al. Obstetric outcomes after transfer of vitrified blastocysts. Hum Reprod 2010; 25: 1699-1707. [DOI:10.1093/humrep/deq117]
4. Surani MA. Imprinting and the initiation of gene silencing in the germ line. Cell 1998; 93: 309-312. [DOI:10.1016/S0092-8674(00)81156-3]
5. Morison IM, Ramsay JP, Spencer HG. A census of mammalian imprinting. Trends Genet 2005; 21: 457-465. [DOI:10.1016/j.tig.2005.06.008]
6. Mayer W, Hemberger M, Frank HG, Grummer R, Winterhager E, Kaufmann P, et al. Expression of the imprinted genes MEST/Mest in human and murine placenta suggests a role in angiogenesis. Dev Dyn 2000; 217: 1-10. https://doi.org/10.1002/(SICI)1097-0177(200001)217:1<1::AID-DVDY1>3.0.CO;2-4 [DOI:10.1002/(SICI)1097-0177(200001)217:13.0.CO;2-4]
7. Nishita Y, Yoshida I, Sado T, Takagi N. Genomic imprinting and chromosomal localization of the human MEST gene. Genomics 1996; 36: 539-542. [DOI:10.1006/geno.1996.0502]
8. Kaneko-Ishino T, Kuroiwa Y, Miyoshi N, Kohda T, Suzuki R, Yokoyama M, et al. Peg1/Mest imprinted gene on chromosome 6 identified by cDNA subtraction hybridization. Nat Genet 1995; 11: 52-59. [DOI:10.1038/ng0995-52]
9. Kobayashi S, Kohda T, Miyoshi N, Kuroiwa Y, Aisaka K, Tsutsumi O, et al. Human PEG1/MEST, an imprinted gene on chromosome 7. Hum Mol Genet 1997; 6: 781-786. [DOI:10.1093/hmg/6.5.781]
10. Huntriss JD, Hemmings KE, Hinkins M, Rutherford A, Sturmey RG, Elder K, et al. Variable imprinting of the MEST gene in human preimplantation embryos. Eur J Hum Genet 2013; 21: 40-47. [DOI:10.1038/ejhg.2012.102]
11. McMinn J, Wei M, Sadovsky Y, Thaker HM, Tycko B. Imprinting of PEG1/MEST isoform 2 in human placenta. Placenta 2006; 27: 119-126. [DOI:10.1016/j.placenta.2004.12.003]
12. Ryan AK, Rosenfeld MG. POU domain family values: flexibility, partnerships, and developmental codes. Genes Dev 1997; 11: 1207-1225. [DOI:10.1101/gad.11.10.1207]
13. Ovitt CE, Scholer HR. The molecular biology of Oct-4 in the early mouse embryo. Mol Hum Reprod 1998; 4: 1021-1031. [DOI:10.1093/molehr/4.11.1021]
14. Scholer HR, Dressler GR, Balling R, Rohdewohld H, Gruss P. Oct-4 :a germline-specific transcription factor mapping to the mouse t-complex. EMBO J 1990; 9: 2185-2195.
15. Dietrich JE, Hiiragi T. Stochastic patterning in the mouse pre-implantation embryo. Development 2007; 134: 4219-4231. [DOI:10.1242/dev.003798]
16. Kehler J, Tolkunova E, Koschorz B, Pesce M, Gentile L, Boiani M, et al. Oct4 is required for primordial germ cell survival. EMBO Rep 2004; 5: 1078-1083. [DOI:10.1038/sj.embor.7400279]
17. Foygel K, Choi B, Jun S, Leong DE, Lee A, Wong CC, et al. A novel and critical role for Oct4 as a regulator of the maternal-embryonic transition. PLoS One 2008; 3: e4109. [DOI:10.1371/journal.pone.0004109]
18. Gardner DK, Lane M, Stevens J, Schoolcraft EB. Blastocyst score affects implantation and pregnancy outcome: toward a single blastocyst transfer. Fertil Steril 2000; 73: 1155-1158. [DOI:10.1016/S0015-0282(00)00518-5]
19. Alipour H, Eftekhari-Yazdi P, Rastgarnia A, Baghaban Eslaminejad MR, Akbarpour M. Effect of LH Treated Ovine Oviductal Epithelial Cell Co- Culture System on Murine Pre-Embryo Development. Iran J Fertil Steril 2008; 2: 131-138.
20. Marshall OJ. PerlPrimer: cross-platform, graphical primer design for standard, bisulphite and real-time PCR. Bioinformatics 2004; 20: 2471-2472. [DOI:10.1093/bioinformatics/bth254]
21. Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 2002; 30: e36. [DOI:10.1093/nar/30.9.e36]
22. Moore K, Bonilla AQ. Cryopreservation of Mammalian Embryos: The State of the Art. Annu Rev Biomed Sci 2006; 8: 19-32.
23. Kader A, Agarwal A, Abdelrazik H, Sharma RK, Ahmady A, Falcone T. Evaluation of post-thaw DNA integrity of mouse blastocysts after ultrarapid and slow freezing. Fertil Steril 2009; 91: 2087-2094. [DOI:10.1016/j.fertnstert.2008.04.049]
24. Al-Hasani S, Ozmen B, Koutlaki N, Schoepper B, Diedrich K, Schultze-Mosgau A. Three years of routine vitrification of human zygotes: is it still fair to advocate slow-rate freezing? Reprod Biomed Online 2007; 14: 288-293. [DOI:10.1016/S1472-6483(10)60869-3]
25. Dhali A, Anchamparuthy VM, Butler SP, Pearson RE, Mullarky IK, Gwazdauskas FC. Effect of droplet vitrification on development competence ,actin cytoskeletal integrity and gene expression in in vitro cultured mouse embryos. Theriogenology 2009; 71: 1408-1416. [DOI:10.1016/j.theriogenology.2009.01.011]
26. Aksu DA, Agca C, Aksu S, Bagis H, Akkoc T, Caputcu AT, et al. Gene expression profiles of vitrified in vitro- and in vivo-derived bovine blastocysts. Mol Reprod Dev 2012; 79: 613-625. [DOI:10.1002/mrd.22068]
27. Sudano MJ, Caixeta ES, Paschoal DM, Rascado TS, Crocomo LF, Maziero RR, et al. Global gene expression patterns of fresh and vitrified in vitro-produced bovine blastocysts. Reprod Fertil Dev 2012; 25: 187-188. [DOI:10.1071/RDv25n1Ab80]
28. Feinberg AP. Genomic imprinting and gene activation in cancer. Nat Genet 1993; 4: 110-113. [DOI:10.1038/ng0693-110]
29. Lefebvre L, Viville S, Barton SC, Ishino F, Keverne EB, Surani MA. Abnormal maternal behaviour and growth retardation associated with loss of the imprinted gene Mest. Nat Genet 1998; 20: 163-169. [DOI:10.1038/2464]
30. Lui JC, Finkielstain GP, Barnes KM, Baron J. An imprinted gene network that controls mammalian somatic growth is down-regulated during postnatal growth deceleration in multiple organs. Am J Physiol Regul Integr Comp Physiol 2008; 295: 189-196. [DOI:10.1152/ajpregu.00182.2008]
31. Nichols J, Zevnik B, Anastassiadis K, Niwa H, Klewe-Nebenius D, Chambers I, et al. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 1998; 95: 379-391. [DOI:10.1016/S0092-8674(00)81769-9]
32. Yeom YI, Fuhrmann G, Ovitt CE, Brehm A, Ohbo K, Gross M, et al. Germline regulatory element of Oct-4 specific for the totipotent cycle of embryonal cells. Development 1996; 122: 881-894.
33. Lenardo MJ, Staudt L, Robbins P, Kuang A, Mulligan RC, Baltimore D. Repression of the IgH enhancer in teratocarcinoma cells associated with a novel octamer factor. Science 1989; 243: 544-546. [DOI:10.1126/science.2536195]
34. Scholer HR, Hatzopoulos AK, Balling R, Suzuki N, Gruss P. A family of octamer-specific proteins present during mouse embryogenesis: evidence for germline-specific expression of an Oct factor. EMBO J 1989; 8: 2543-2550.
35. Zhu D, Zhang J, Cao S, Zhang J, Heng BC, Huang M, et al. Vitrified-warmed blastocyst transfer cycles yield higher pregnancy and implantation rates compared with fresh blastocyst transfer cycles --time for a new embryo transfer strategy? Fertil Steril 2011; 95: 1691-1695. [DOI:10.1016/j.fertnstert.2011.01.022]
36. Zhao XM, Du WH, Hao HS, Wang D, Qin T, Liu Y, et al. Effect of vitrification on promoter methylation and the expression of pluripotency and differentiation genes in mouse blastocysts. Mol Reprod Dev 2012; 79: 445-450. [DOI:10.1002/mrd.22052]
37. Desai N, Xu J, Tsulaia T, Szeptycki-Lawson J, AbdelHafez F, Goldfarb J, et al. Vitrification of mouse embryo-derived ICM cells: a tool for preserving embryonic stem cell potential? J Assist Reprod Genet 2011; 28: 93-99. [DOI:10.1007/s10815-010-9500-x]

Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Designed & Developed by : Yektaweb