Original Articles

Comparison of Gene Expression Profiles in Human Germinal Vesicle Before and After Cytoplasmic Transfer From Mature Oocytes in Iranian Infertile Couples

Abstract

Objective: To evaluate the effect of cytoplasm transfer from mature oocytes to germinal vesicle(GV)s on promoting the maturation of cytoplasm of GV at the mRNA level.
Materials and methods: Sixty six in vitro fertilization (IVF) operations between June 2012 and November 2013 were included in this study. Totally 120 GVs were obtained. Normal GVs were categorized into 3 groups (n = 40) randomly: the first comprised oocytes that did not receive the cytoplasm of mature oocytes; the second group comprised oocytes that did not receive the cytoplasm of mature oocytes but were incubated for 24 h; and the third group comprised oocytes that received 10-15% the cytoplasm of mature oocytes and were then incubated for 24 h. Each group was separately analyzed by quantitative polymerase chain reaction (qPCR) and the expression levels of selected genes were assessed.
Results: The expression levels of genes involved in the cytoplasmic maturity, and energy-producing mitochondria were significantly higher in the pooled oocytes of 2nd control group than those of the 1st control and intervention groups (p < 0.001). The genes involved in the meiosis, spindle check point, DNA repairing and cell cycle checkpoint did not have any expression in the 1st and intervention groups; however, these genes were expressed in the 2nd group, significantly. In the 2nd group, the highest expression level was observed for genes involved in the DNA repairing and cell cycle checkpoint. In the intervention group, none of the genes were expressed except for energy-producing mitochondria gene; even in this case, the expression level of this gene in this group of oocytes was significantly lower than that in other groups (p < 0.001). After 24 h meiosis assumption was significantly higher in the third group than in the second group (95% vs. 68%, p < 0.001).
Conclusion: The cytoplasm transfer technique is not effective in cytoplasmic maturity of the recipient GV oocytes. In contrast, 24-hr in-vitro culture is associated with increased expression of studied genes in GVs.

Virant-Klun I, Knez K, Tomazevic T, Skutella T. Gene expression profiling of human oocytes developed and matured in vivo or in vitro. Biomed Res Int 2013;2013:879489.

Wells D, Bermudez MG, Steuerwald N, Thornhill AR, Walker DL, Malter H, et al. Expression of genes regulating chromosome segregation, the cell cycle and apoptosis during human preimplantation development. Hum Reprod 2005;20 :1339-48.

Assou S, Anahory T, Pantesco V, Le Carrour T, Pellestor F, Klein B, et al. The human cumulus--oocyte complex gene-expression profile. Hum Reprod 2006; 21 : 1705-19.

Gasca S, Pellestor F, Assou S, Loup V, Anahory T, Dechaud H, et al. Identifying new human oocyte marker genes: a microarray approach. Reprod Biomed Online 2007;14 :175-83.

Malter HE, Schimmel T, Cohen J. Development of a mouse model for ooplasmic transplantation. Fertility and Sterility 2002;78:S83.

Malter HE, Cohen J. Ooplasmic transfer: animal models assist human studies. Reprod BioMed Online 2002;5 :26-35.

Flood JT, Chillik CF, van Uem JF, Iritani A, Hodgen GD. Ooplasmic transfusion: prophase germinal vesicle oocytes made developmentally competent by microinjection of metaphase II egg cytoplasm. Fertil Steril 1990;53: 1049-54.

Cohen J, Scott R, Schimmel T, Levron J, Willadsen S. Birth of infant after transfer of anucleate donor oocyte cytoplasm into recipient eggs. Lancet 1997;350: 186-7.

Dale B, Wilding M, Botta G, Rasile M, Marino M, Di Matteo L, et al. Pregnancy after cytoplasmic transfer in a couple suffering from idiopathic infertility. Hum Reprod. 2001;16:1469-72.

Barritt JA, Tomkin G, Sable DB, Cohen J. Effects of cytoplasmic transfer on embryo quality are post-genomic. Fertil Steril 2001;76 :S6

Lanzendorf SE, Mayer JF, Toner J, Oehninger S, Saffan DS, Muasher S. Pregnancy following transfer of ooplasm from cryopreserved-thawed donor oocytes into recipient oocytes. Fertil Steril 1999;71:575-7.

Levy R, Elder K, Menezo Y. Cytoplasmic transfer in oocytes: biochemical aspects. Hum Reprod Update 2004;10:241-50.

Ye H, Huang GN, Zeng PH, Pei L. IVF/ICSI outcomes between cycles with luteal estradiol (E2) pre-treatment before GnRH antagonist protocol and standard long GnRH agonist protocol . J Assist Reprod Genet 2009;26 :105-11.

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.

Hsieh RH, Au HK, Yeh TS, Chang SJ, Cheng YF, Tzeng CR. Decreased expression of mitochondrial genes in human unfertilized oocytes and arrested embryos. Fertil Steril 2004; 81 Suppl 1:912-8.

Lee SH, Lee S, Jun HS, Jeong HJ, Cha WT, Cho YS, et al. Expression of the mitochondrial ATPase6 gene and Tfam in Down syndrome. Mol Cells 2003;15: 181-5.

Lee SH, Han JH, Cho SW, Cha KE, Park SE, Cha KY. Mitochondrial ATPase 6 gene expression in unfertilized oocytes and cleavage-stage embryos. Fertil Steril 2000;73:1001-5.

Barritt J, Willadsen S, Brenner C, Cohen J. Cytoplasmic transfer in assisted reproduction. Hum Reprod Update 2001;7 :428-35.

Dekel N. Cellular, biochemical and molecular mechanisms regulating oocyte maturation. Mol Cell Endocrinol 2005;234: 19-25.

Wei LN, Liang XY, Fang C, Zhang MF. Abnormal expression of growth differentiation factor 9 and bone morphogenetic protein 15 in stimulated oocytes during maturation from women with polycystic ovary syndrome. Fertil Steril 2011;96: 464-8.

Paulini F, Melo EO. The role of oocyte-secreted factors GDF9 and BMP15 in follicular development and oogenesis. Reprod Domest Anim 2011;46: 354-61.

Knight PG, Glister C. TGF-beta superfamily members and ovarian follicle development. Reproduction 2006;132: 191-206.

Bebbere D, Bogliolo L, Ariu F, Fois S, Leoni GG, Tore S, et al. Expression pattern of zygote arrest 1 (ZAR1), maternal antigen that embryo requires (MATER), growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) genes in ovine oocytes and in vitro-produced preimplantation embryos. Reprod Fertil Dev 2008;20: 908-15.

Zhao SY, Qiao J, Chen YJ, Liu P, Li J, Yan J. Expression of growth differentiation factor-9 and bone morphogenetic protein-15 in oocytes and cumulus granulosa cells of patients with polycystic ovary syndrome. Fertil Steril 2010;94 : 261-7.

Goud AP, Goud PT, Van Oostveldt P, Diamond MP, Dhont M. Dynamic changes in microtubular cytoskeleton of human postmature oocytes revert after ooplasm transfer. Fertil Steril 2004;81:323-31.

Tibbetts RS, Cortez D, Brumbaugh KM, Scully R, Livingston D, Elledge SJ, et al. Functional interactions between BRCA1 and the checkpoint kinase ATR during genotoxic stress. Genes Dev 2000;14 :2989-3002.

Garcia-Cruz R, Roig I, Robles P, Scherthan H, Garcia Caldes M. ATR, BRCA1 and gammaH2AX localize to unsynapsed chromosomes at the pachytene stage in human oocytes. Reprod Biomed Online 2009;18:

-44.

Verhagen AM, Coulson EJ, Vaux DL. Inhibitor of apoptosis proteins and their relatives: IAPs and other BIRPs. Genome Biol 2001;2: REVIEWS 3009.

Hawes SM, Sapienza C, Latham KE. Ooplasmic donation in humans: the potential for epigenic modifications. Hum Reprod 2002;17:850-2.

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IssueVol 10, No 2 (June 2016) QRcode
SectionOriginal Articles
Keywords
Gene Expression Cytoplasmic Transfer Oocyte Maturity

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How to Cite
1.
Hoseini FS, Salsabili N, Akbari-Asbagh F, Aflatoonian R, Aghaee-Bakhtiari SH. Comparison of Gene Expression Profiles in Human Germinal Vesicle Before and After Cytoplasmic Transfer From Mature Oocytes in Iranian Infertile Couples. J Family Reprod Health. 2016;10(2):71-79.