International Journal of
Reproductive Biomedicine
Tue, Apr 23, 2024
[Archive]
Volume 14, Issue 8 (8-2016)
IJRM 2016, 14(8): 511-518 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Majidi Gharenaz N, Movahedin M, Mazaheri Z, Pour beiranvand S. Alternation of apoptotic and implanting genes expression of mouse embryos after re-vitrification. IJRM 2016; 14 (8) :511-518
URL: http://ijrm.ir/article-1-775-en.html
URL: http://ijrm.ir/article-1-775-en.html
1- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran , mansoure@modares.ac.ir
2- Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran , mansoure@modares.ac.ir
Abstract: (2465 Views)
Background: Nowadays, oocytes and embryos vitrification has become a routine technique. Based on clinical judgment, re-vitrification maybe required. But little is known about re-vitrification impact on genes expression.
Objective: The impact of re-vitrification on apoptotic and implanting genes, Bax, Bcl-2 and ErbB4, at compaction stage embryos were evaluated in this study.
Materials and Methods: In this experimental study, 8 cell embryos (n=240) were collected from female mature mice, 60-62 hr post HCG injection. The embryos were divided randomly to 3 groups included: fresh (n=80), vitrified at 8 cell stage (n=80), vitrified at 8 cell stage thawed and re-vitrified at compaction stage (n=80). Embryos were vitrified by using cryolock, (open system) described by Kuwayama. Q-PCR was used to examine the expression of Bax, Bcl2 ErbB4 genes in derived blastocysts.
Results: Our result showed that expanded blastocyst rate was similar between vitrified and re-vitrified groups, while re-vitrified embryos showed significant decrease in expanded blastocyst rate comparing with fresh embryos (p=0.03). In addition, significant difference was observed on apoptotic gene expression when comparing re-vitrified and fresh embryos (p=0.004), however expression of Bax and Bcl-2 (apoptotic) genes didn't demonstrate a significant difference between re-vitrified and vitrified groups. The expression rate of ErbB4, an implantation gene was decreased in re-vitrified embryos comparing with fresh embryos (p=0.003), but it was similar between re-vitrified and vitrified embryos.
Conclusion: Re-vitrification can alter the expression of Bax, Bcl-2 and ErbB4 genes and developmental rate of mouse embryos in compaction stage.
Objective: The impact of re-vitrification on apoptotic and implanting genes, Bax, Bcl-2 and ErbB4, at compaction stage embryos were evaluated in this study.
Materials and Methods: In this experimental study, 8 cell embryos (n=240) were collected from female mature mice, 60-62 hr post HCG injection. The embryos were divided randomly to 3 groups included: fresh (n=80), vitrified at 8 cell stage (n=80), vitrified at 8 cell stage thawed and re-vitrified at compaction stage (n=80). Embryos were vitrified by using cryolock, (open system) described by Kuwayama. Q-PCR was used to examine the expression of Bax, Bcl2 ErbB4 genes in derived blastocysts.
Results: Our result showed that expanded blastocyst rate was similar between vitrified and re-vitrified groups, while re-vitrified embryos showed significant decrease in expanded blastocyst rate comparing with fresh embryos (p=0.03). In addition, significant difference was observed on apoptotic gene expression when comparing re-vitrified and fresh embryos (p=0.004), however expression of Bax and Bcl-2 (apoptotic) genes didn't demonstrate a significant difference between re-vitrified and vitrified groups. The expression rate of ErbB4, an implantation gene was decreased in re-vitrified embryos comparing with fresh embryos (p=0.003), but it was similar between re-vitrified and vitrified embryos.
Conclusion: Re-vitrification can alter the expression of Bax, Bcl-2 and ErbB4 genes and developmental rate of mouse embryos in compaction stage.
Type of Study: Original Article |
References
1. Liebermann J, Dietl J, Vanderzwalmen P, Tucker MJ. Recent developments in human oocyte, embryo and blastocyst vitrification: where are we now? Reprod Biomed Online 2003; 7: 623-633. [DOI:10.1016/S1472-6483(10)62084-6]
2. Vajta G, Nagy ZP. Are programmable freezers still needed in the embryo laboratory? Review on vitrification. Reprod Biomed Online 2006; 12: 779-796. [DOI:10.1016/S1472-6483(10)61091-7]
3. Wennerholm U-B, Söderström-Anttila V, Bergh C, Aittomäki K, Hazekamp J, Nygren K-G, et al. Children born after cryopreservation of embryos or oocytes: a systematic review of outcome data. Hum Reprod 2009; 24: 2158-2172. [DOI:10.1093/humrep/dep125]
4. Bhattacharya S, Kamath MS. Reducing multiple births in assisted reproduction technology. Best Pract Res Clin Obstet Gynaecol 2014; 28: 191-199. [DOI:10.1016/j.bpobgyn.2013.11.005]
5. Belva F, Henriet S, Van den Abbeel E, Camus M, Devroey P, Van der Elst J, et al. Neonatal outcome of 937 children born after transfer of cryopreserved embryos obtained by ICSI and IVF and comparison with outcome data of fresh ICSI and IVF cycles. Hum Reprod 2008; 23: 2227-2238. [DOI:10.1093/humrep/den254]
6. Pelkonen S, Koivunen R, Gissler M, Nuojua-Huttunen S, Suikkari A-M, Hyden-Granskog C, et al. Perinatal outcome of children born after frozen and fresh embryo transfer: the Finnish cohort study 1995-2006. Hum Reprod 2010; 25: 914-923. [DOI:10.1093/humrep/dep477]
7. Roque M, Lattes K, Serra S, Solà I, Geber S, Carreras R, et al. Fresh embryo transfer versus frozen embryo transfer in in vitro fertilization cycles: a systematic review and meta-analysis. Fertil Steril 2013; 99: 156-162. [DOI:10.1016/j.fertnstert.2012.09.003]
8. Bedoschi G, Oktay K. Current approach to fertility preservation by embryo cryopreservation. Fertil Steril 2013; 99: 1496-1502. [DOI:10.1016/j.fertnstert.2013.03.020]
9. Elnahas A, Alcolak E, Marar EA, Elnahas T, Elnahas K, Palapelas V, et al. Vitrification of human oocytes and different development stages of embryos: An overview. Middle East Fertil Soc J 2010; 15: 2-9. [DOI:10.1016/j.mefs.2010.03.013]
10. Matsumoto H, Jiang J, Tanaka T, Sasada H, Sato E. Vitrification of large quantities of immature bovine oocytes using nylon mesh. Cryobiology 2001; 42: 139-144. [DOI:10.1006/cryo.2001.2309]
11. Farhat M, Zentner B-S, Lossos F, Bdolah Y, Holtzer H, Hurwitz A. Successful pregnancy following replacement of embryos previously refrozen at blastocyst stage: case report. Hum Reprod 2001; 16: 337-339. [DOI:10.1093/humrep/16.2.337]
12. Koutlaki N, Schoepper B, Maroulis G, Diedrich K, Al-Hasani S. Human oocyte cryopreservation: past, present and future. Reprod Biomed Online 2006; 13: 427-436. [DOI:10.1016/S1472-6483(10)61449-6]
13. El-Toukhy T, Khalaf Y, Al-Darazi K, Andritsos V, Taylor A, Braude P. Effect of blastomere loss on the outcome of frozen embryo replacement cycles. Fertil Steril 2003; 79: 1106-1111. [DOI:10.1016/S0015-0282(03)00072-4]
14. Inaba Y, Miyashita S, Somfai T, Geshi M, Matoba S, Dochi O, et al. Cryopreservation method affects DNA fragmentation in trophectoderm and the speed of re-expansion in bovine blastocysts. Cryobiology 2016; 72: 86-92. [DOI:10.1016/j.cryobiol.2016.03.006]
15. Park SY, Kim EY, Cui XS, Tae JC, Lee WD, Kim NH, et al. Increase in DNA fragmentation and apoptosis-related gene expression in frozen-thawed bovine blastocysts. Zygote 2006; 14: 125-131. [DOI:10.1017/S0967199406003649]
16. Dhali A, Anchamparuthy V, Butler S, Pearson R, Mullarky I, Gwazdauskas F. Gene expression and development of mouse zygotes following droplet vitrification. Theriogenology 2007; 68: 1292-1298. [DOI:10.1016/j.theriogenology.2007.08.030]
17. Anchamparuthy V, Dhali A, Butler S, Pearson R, Jiang H, Gwazdauskas F. Effect of Vitrification of Zygotes in Nylon Mesh on the Differential Expression of Apoptotic Genes in Mouse Embryos. World J Dairy Food Sci 2007; 2: 54-61.
18. Dhali A, Anchamparuthy V, Butler S, Pearson R, Mullarky I, Gwazdauskas F. 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]
19. Yang MY, Rajamahendran R. Expression of Bcl-2 and Bax proteins in relation to quality of bovine oocytes and embryos produced in vitro. Anim Reprod Sci 2002; 70: 159-169. [DOI:10.1016/S0378-4320(01)00186-5]
20. Cha J, Sun X, Dey SK. Mechanisms of implantation: strategies for successful pregnancy. Nature Med 2012; 18: 1754-1767. [DOI:10.1038/nm.3012]
21. Davidson LM, Coward K. Molecular mechanisms of membrane interaction at implantation. Birth Defects Res Part C Embryo Today 2016; 108: 19-32. [DOI:10.1002/bdrc.21122]
22. Estes SJ, Laky DC, Hoover LM, Smith SE, Schinfeld JS, Somkuti SG. Successful pregnancy resulting from cryopreserved pronuclear and cleaved embryos thawed and cultured to blastocysts, refrozen and transferred. A case report. J Reprod Med 2003; 48: 46-48.
23. Smith LK, Roots EH, Dorsett MJO. Live birth of a normal healthy baby after a frozen embryo transfer with blastocysts that were frozen and thawed twice. Fertil Steril 2005; 83: 198-200. [DOI:10.1016/j.fertnstert.2004.07.953]
24. Sheehan CB, Lane M, Gardner DK. The CryoLoop facilitates re-vitrification of embryos at four successive stages of development without impairing embryo growth. Hum Reprod 2006; 21: 2978-2984. [DOI:10.1093/humrep/del253]
25. Shin MR, Choi HW, Kim MK, Lee SH, Lee HS, Lim CK. In vitro development and gene expression of frozen-thawed 8-cell stage mouse embryos following slow freezing or vitrification. Clin Exp Reprod Med 2011; 38: 203-209. [DOI:10.5653/cerm.2011.38.4.203]
26. Kuwayama M, Vajta G, Ieda S, Kato O. Comparison of open and closed methods for vitrification of human embryos and the elimination of potential contamination. Reprod Biomed Online 2005; 11: 608-614. [DOI:10.1016/S1472-6483(10)61169-8]
27. Fathi R, Valojerdi MR, Yazdi PE, Ebrahimi B, Alipour H, Hassani F. Development of 4-cell mouse embryos after re-vitrification. Cryobiology 2012; 64: 23-26. [DOI:10.1016/j.cryobiol.2011.11.003]
28. Rall W. Factors affecting the survival of mouse embryos cryopreserved by vitrification. Cryobiology 1987; 24: 387-402. [DOI:10.1016/0011-2240(87)90042-3]
29. Lane M, Maybach JM, Gardner DK. Addition of ascorbate during cryopreservation stimulates subsequent embryo development. Hum Reprod 2002; 17: 2686-2693. [DOI:10.1093/humrep/17.10.2686]
30. Halliwell B, Gutteridge JM. Free radicals in biology and medicine. Oxford University Press, USA; 2015. [DOI:10.1093/acprof:oso/9780198717478.001.0001]
31. El-Gayar M, Gauly M, Holtz W. In vitro and in vivo survival of mouse blastocysts after repeated vitrification with the open pulled straw (OPS) method. Cryoletters 2010; 31: 454-459.
32. Toner M, Cravalho E, Stachecki J, Fitzgerald T, Tompkins R, Yarmush M, et al. Nonequilibrium freezing of one-cell mouse embryos. Membrane integrity and developmental potential. Biophys J 1993; 64: 1908-1021. [DOI:10.1016/S0006-3495(93)81562-5]
33. Ahn HJ, Sohn IP, Kwon HC, Jo DH, Park YD, Min CK. Characteristics of the cell membrane fluidity, actin fibers, and mitochondrial dysfunctions of frozen‐thawed two‐cell mouse embryos. Mol Reprod Dev 2002; 61: 466-476. [DOI:10.1002/mrd.10040]
34. Lane M, Lyons EA, Bavister BD. Cryopreservation reduces the ability of hamster 2-cell embryos to regulate intracellular pH. Hum Reprod 2000; 15: 389-394. [DOI:10.1093/humrep/15.2.389]
35. Kasai M, Ito K, Edashige K. Morphological appearance of the cryopreserved mouse blastocyst as a tool to identify the type of cryoinjury. Hum Reprod 2002; 17: 1863-1874. [DOI:10.1093/humrep/17.7.1863]
36. Rajaei F, Karja N, Agung B, Wongsrikeao P, Taniguchi M, Murakami M, et al. Analysis of DNA fragmentation of porcine embryos exposed to cryoprotectants. Reprod Domest Anim 2005; 40: 429-432. [DOI:10.1111/j.1439-0531.2005.00585.x]
37. Paria B, Das S, Andrews G, Dey S. Expression of the epidermal growth factor receptor gene is regulated in mouse blastocysts during delayed implantation. Proc Nat Acad Sci 1993; 90: 55-59. [DOI:10.1073/pnas.90.1.55]
38. Valles CS, Domínguez F. Embryo-endometrial interaction. Chang Gung Med J 2006; 29: 9-14.
39. Kaneko Y, Lecce L, Day ML, Murphy CR. β1 and β3 integrins disassemble from basal focal adhesions and β3 integrin is later localised to the apical plasma membrane of rat uterine luminal epithelial cells at the time of implantation. Reprod Fertil Dev 2011; 23: 481-495. [DOI:10.1071/RD10211]
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
