Top

Journal of Assisted Reproduction and Genetics

Published in:

01-11-2014 | Epigenetics

Altered imprinted gene expression and methylation patterns in mid-gestation aborted cloned porcine fetuses and placentas

Authors: Xiaoyang Zhang, Dongxu Wang, Yang Han, Feifei Duan, Qinyan Lv, Zhanjun Li

Published in: Journal of Assisted Reproduction and Genetics | Issue 11/2014

Abstract

Purpose

To determine the expression patterns of imprinted genes and their methylation status in aborted cloned porcine fetuses and placentas.

Methods

RNA and DNA were prepared from fetuses and placentas that were produced by SCNT and controls from artificial insemination. The expression of 18 imprinted genes was determined by quantitative real-time PCR (q-PCR). Bisulfite sequencing PCR (BSP) was conducted to determine the methylation status of PRE-1 short interspersed repetitive element (SINE), satellite DNA and H19 differentially methylated region 3 (DMR3).

Results

The weight, imprinted gene expression and genome-wide DNA methylation patterns were compared between the mid-gestation aborted and normal control samples. The results showed hypermethylation of PRE-1 and satellite sequences, the aberrant expression of imprinted genes, and the hypomethylation of H19 DMR3 occurred in mid-gestation aborted fetuses and placentas.

Conclusions

Cloned pigs generated by somatic cell nuclear transfer (SCNT) showed a greater ratio of early abortion during mid-gestation than did normal controls because of the incomplete epigenetic reprogramming of the donor cells. Altered expression of imprinted genes and the hypermethylation profile of the repetitive regions (PRE-1 and satellite DNA) may be associated with defective development and early abortion of cloned pigs, emphasizing the importance of epigenetics during pregnancy and implications thereof for patient-specific embryonic stem cells for human therapeutic cloning and improvement of human assisted reproduction.

Available only for authorised users

Wilmut I, Beaujean N, de Sousa PA, et al. Somatic cell nuclear transfer. Nature. 2002;419(6907):583–6. doi:10.1038/nature01079.PubMedCrossRef

Yang X, Smith SL, Tian XC, et al. Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning. Nat Genet. 2007;39(3):295–302. doi:10.1038/ng1973.PubMedCrossRef

Chae JI, Lee KS, Kim DJ, et al. Abnormal gene expression in extraembryonic tissue from cloned porcine embryos. Theriogenology. 2009;71(2):323–33. doi:10.1016/j.theriogenology.2008.07.025.PubMedCrossRef

Chae JI, Yu K, Cho SK, et al. Aberrant expression of developmentally important signaling molecules in cloned porcine extraembryonic tissues. Proteomics. 2008;8(13):2724–34. doi:10.1002/pmic.200701134.PubMedCrossRef

Young LE, Fairburn HR. Improving the safety of embryo technologies: possible role of genomic imprinting. Theriogenology. 2000;53(2):627–48.PubMedCrossRef

Jiang L, Jobst P, Lai L, et al. Expression levels of growth-regulating imprinted genes in cloned piglets. Cloning Stem Cells. 2007;9(1):97–106. doi:10.1089/clo.2006.0041.PubMedCrossRef

Shi W, Dirim F, Wolf E, et al. Methylation reprogramming and chromosomal aneuploidy in in vivo fertilized and cloned rabbit preimplantation embryos. Biol Reprod. 2004;71(1):340–7.PubMedCrossRef

Kang YK, Koo DB, Park JS, et al. Aberrant methylation of donor genome in cloned bovine embryos. Nat Genet. 2001;28(2):173–7. doi:10.1038/88903.PubMedCrossRef

Tajima K, Enishi O, Amari M, et al. PCR detection of DNAs of animal origin in feed by primers based on sequences of short and long interspersed repetitive elements. Biosci Biotechnol Biochem. 2002;66(10):2247–50.PubMedCrossRef

10.

Harumi T, Kimura M, Yasue H. Survey on swine SINEs (PRE‐1) as candidates for SSCP markers in genetic linkage analysis. Anim Genet. 1995;26(6):403–6.PubMedCrossRef

11.

Park C-H, Kim H-S, Lee S-G, et al. Methylation status of differentially methylated regions at <i> Igf2/H19</i> locus in porcine gametes and preimplantation embryos. Genomics. 2009;93(2):179–86.PubMedCrossRef

12.

Whitelaw N, Bhattacharya S, Hoad G, Horgan GW, Hamilton M, Haggarty P. Epigenetic status in the offspring of spontaneous and assisted conception. Hum Reprod. 2014. doi:10.1093/humrep/deu094.

13.

Huang Y, Ouyang H, Yu H, et al. Efficiency of porcine somatic cell nuclear transfer - a retrospective study of factors related to embryo recipient and embryos transferred. Biol Open. 2013;2(11):1223–8. doi:10.1242/bio.20135983.PubMedCentralPubMedCrossRef

14.

Lai L, Prather RS. Production of cloned pigs by using somatic cells as donors. Cloning & Stem Cells. 2003;5(4):233–41.CrossRef

15.

Kang YK, Koo DB, Park JS, et al. Typical demethylation events in cloned pig embryos. Clues on species-specific differences in epigenetic reprogramming of a cloned donor genome. J Biol Chem. 2001;276(43):39980–4. doi:10.1074/jbc.M106516200.PubMedCrossRef

16.

Frank D, Fortino W, Clark L, et al. Placental overgrowth in mice lacking the imprinted gene Ipl. Proc Natl Acad Sci U S A. 2002;99(11):7490–5. doi:10.1073/pnas.122039999.PubMedCentralPubMedCrossRef

17.

Wei Y, Zhu J, Huan Y, et al. Aberrant expression and methylation status of putatively imprinted genes in placenta of cloned piglets. Cell Reprogram. 2010;12(2):213–22. doi:10.1089/cell.2009.0090.PubMedCrossRef

18.

Su JM, Yang B, Wang YS, et al. Expression and methylation status of imprinted genes in placentas of deceased and live cloned transgenic calves. Theriogenology. 2011;75(7):1346–59. doi:10.1016/j.theriogenology.2010.11.045.PubMedCrossRef

19.

Amilcar Cordeiro APN, Filipa Carvalho, Carla Ramalho & Sofia Dória. Relevance of genomic imprinting in intrauterine human growth expression of CDKN1C, H19, IGF2, KCNQ1 and PHLDA2 imprinted genes. J Assist Reprod Genet. 2014. doi:10.1007/s10815-014-0278-0.

20.

Mayer W, Niveleau A, Walter J, et al. Demethylation of the zygotic paternal genome. Nature. 2000;403(6769):501–2. doi:10.1038/35000654.PubMedCrossRef

21.

Market-Velker BA, Fernandes AD, Mann MR. Side-by-side comparison of five commercial media systems in a mouse model: suboptimal in vitro culture interferes with imprint maintenance. Biol Reprod. 2010;83(6):938–50. doi:10.1095/biolreprod.110.085480.PubMedCrossRef

22.

Hall JG. Genomic imprinting: review and relevance to human diseases. Am J Hum Genet. 1990;46(5):857–73.PubMedCentralPubMed

Title: Altered imprinted gene expression and methylation patterns in mid-gestation aborted cloned porcine fetuses and placentas
Authors: Xiaoyang Zhang
Dongxu Wang
Yang Han
Feifei Duan
Qinyan Lv
Zhanjun Li
Publication date: 01-11-2014
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 11/2014
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-014-0320-2

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Altered imprinted gene expression and methylation patterns in mid-gestation aborted cloned porcine fetuses and placentas

Abstract

Purpose

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 11/2014

Deliveries of babies with normal health derived from oocytes with smooth endoplasmic reticulum clusters

Treatment history and outcome of 24 deliveries worldwide after autotransplantation of cryopreserved ovarian tissue, including two new Danish deliveries years after autotransplantation

Serum tumor necrosis factor-α, interleukin-6, monocyte chemotactic protein-1 and paraoxonase-1 profiles in women with endometriosis, pcos, or unexplained infertility

Association of rs7260002 of chorionic gonadotrophin β5 with idiopathic recurrent spontaneous abortion in Chinese population

Administration of DHEA augments progesterone production in a woman with low ovarian reserve being transplanted with cryopreserved ovarian tissue

Approaches to oocyte retrieval for advanced reproductive technology cycles planning to utilize in vitro maturation: a review of the many choices to be made