Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes

El Hajj N, Trapphoff T, Linke M, May A, Hansmann T, Kuhtz J, Reifenberg K, Heinzmann J, Niemann H, Daser A, Eichenlaub-Ritter U, et al. (2011)
Epigenetics 6(10): 1176-1188.

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Abstract
To detect rare epigenetic effects associated with assisted reproduction, it is necessary to monitor methylation patterns of developmentally important genes in a few germ cells and individual embryos. Bisulfite treatment degrades DNA and reduces its complexity, rendering methylation analysis from small amounts of DNA extremely challenging. Here we describe a simple approach that allows determining the parent-specific methylation patterns of multiple genes in individual early embryos. Limiting dilution (LD) of bisulfite-treated DNA is combined with independent multiplex PCRs of single DNA target molecules to avoid amplification bias. Using this approach, we compared the methylation status of three imprinted (H19, Snrpn and Igf2r) and one pluripotency-related gene (Oct4) in three different groups of single mouse two-cell embryos. Standard in vitro fertilization of superovulated oocytes and the use of in vitro matured oocytes were not associated with significantly increased rates of stochastic single CpG methylation errors and epimutations (allele methylation errors), when compared with the in vivo produced controls. Similarly, we compared the methylation patterns of two imprinted genes (H19 and Snrpn) in individual mouse 16-cell embryos produced in vivo from superovulated and non-superovulated oocytes and did not observe major between-group differences. Using bovine oocytes and polar bodies as a model, we demonstrate that LD even allows the methylation analysis of multiple genes in single cells.
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El Hajj N, Trapphoff T, Linke M, et al. Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes. Epigenetics. 2011;6(10):1176-1188.
El Hajj, N., Trapphoff, T., Linke, M., May, A., Hansmann, T., Kuhtz, J., Reifenberg, K., et al. (2011). Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes. Epigenetics, 6(10), 1176-1188.
El Hajj, N., Trapphoff, T., Linke, M., May, A., Hansmann, T., Kuhtz, J., Reifenberg, K., Heinzmann, J., Niemann, H., Daser, A., et al. (2011). Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes. Epigenetics 6, 1176-1188.
El Hajj, N., et al., 2011. Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes. Epigenetics, 6(10), p 1176-1188.
N. El Hajj, et al., “Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes”, Epigenetics, vol. 6, 2011, pp. 1176-1188.
El Hajj, N., Trapphoff, T., Linke, M., May, A., Hansmann, T., Kuhtz, J., Reifenberg, K., Heinzmann, J., Niemann, H., Daser, A., Eichenlaub-Ritter, U., Zechner, U., Haaf, T.: Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes. Epigenetics. 6, 1176-1188 (2011).
El Hajj, Nady, Trapphoff, Tom, Linke, Matthias, May, Andreas, Hansmann, Tamara, Kuhtz, Juliane, Reifenberg, Kurt, Heinzmann, Julia, Niemann, Heiner, Daser, Angelika, Eichenlaub-Ritter, Ursula, Zechner, Ulrich, and Haaf, Thomas. “Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes”. Epigenetics 6.10 (2011): 1176-1188.
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Data provided by Europe PubMed Central.

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PMID: 25084396
In vitro maturation of oocytes is not a risk factor for adult metabolic syndrome of mouse offspring.
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PMID: 24405727

38 References

Data provided by Europe PubMed Central.

Epigenetic profile of developmentally important genes in bovine oocytes.
Heinzmann J, Hansmann T, Herrmann D, Wrenzycki C, Zechner U, Haaf T, Niemann H., Mol. Reprod. Dev. 78(3), 2011
PMID: 21290475
Methylation imprints of the imprint control region of the SNRPN-gene in human gametes and preimplantation embryos.
Geuns E, De Rycke M, Van Steirteghem A, Liebaers I., Hum. Mol. Genet. 12(22), 2003
PMID: 14500540
Methylation analysis of KvDMR1 in human oocytes.
Geuns E, Hilven P, Van Steirteghem A, Liebaers I, De Rycke M., J. Med. Genet. 44(2), 2007
PMID: 16950814
Methylation analysis of the intergenic differentially methylated region of DLK1-GTL2 in human.
Geuns E, De Temmerman N, Hilven P, Van Steirteghem A, Liebaers I, De Rycke M., Eur. J. Hum. Genet. 15(3), 2007
PMID: 17213841
Identification of rare DNA variants in mitochondrial disorders with improved array-based sequencing.
Wang W, Shen P, Thiyagarajan S, Lin S, Palm C, Horvath R, Klopstock T, Cutler D, Pique L, Schrijver I, Davis RW, Mindrinos M, Speed TP, Scharfe C., Nucleic Acids Res. 39(1), 2011
PMID: 20843780
Dynamics, stability and inheritance of somatic DNA methylation imprints.
Sontag LB, Lorincz MC, Georg Luebeck E., J. Theor. Biol. 242(4), 2006
PMID: 16806276
Distribution, silencing potential and evolutionary impact of promoter DNA methylation in the human genome.
Weber M, Hellmann I, Stadler MB, Ramos L, Paabo S, Rebhan M, Schubeler D., Nat. Genet. 39(4), 2007
PMID: 17334365
Superovulation alters the expression of imprinted genes in the midgestation mouse placenta.
Fortier AL, Lopes FL, Darricarrere N, Martel J, Trasler JM., Hum. Mol. Genet. 17(11), 2008
PMID: 18287259
Differential effects of culture on imprinted H19 expression in the preimplantation mouse embryo.
Doherty AS, Mann MR, Tremblay KD, Bartolomei MS, Schultz RM., Biol. Reprod. 62(6), 2000
PMID: 10819752
Culture of preimplantation mouse embryos affects fetal development and the expression of imprinted genes.
Khosla S, Dean W, Brown D, Reik W, Feil R., Biol. Reprod. 64(3), 2001
PMID: 11207209
Selective loss of imprinting in the placenta following preimplantation development in culture.
Mann MR, Lee SS, Doherty AS, Verona RI, Nolen LD, Schultz RM, Bartolomei MS., Development 131(15), 2004
PMID: 15240554
Extreme methylation values of imprinted genes in human abortions and stillbirths.
Pliushch G, Schneider E, Weise D, El Hajj N, Tresch A, Seidmann L, Coerdt W, Muller AM, Zechner U, Haaf T., Am. J. Pathol. 176(3), 2010
PMID: 20093482
DNA methylation mapping by tag-modified bisulfite genomic sequencing.
Han W, Cauchi S, Herman JG, Spivack SD., Anal. Biochem. 355(1), 2006
PMID: 16797472

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