Abstract
An association between assisted reproductive technology (ART) and neurobehavioral imprinting disorders has been reported in many studies, and it seems that ART may interfere with imprint reprogramming. However, it has never been explored whether epigenetic errors or imprinting disease susceptibility induced by ART can be inherited transgenerationally. Hence, the aim of this study was to determine the effect of in vitro fertilization and embryo transfer (IVF-ET) on transgenerational inheritance in an inbred mouse model. Mice derived from IVF-ET were outcrossed to wild-type C57BL/6J to obtain their female and male line F2 and F3 generations. Their behavior, morphology, histology, and DNA methylation status at several important differentially methylated regions (DMRs) were analyzed by Morris water maze, hematoxylin and eosin (H&E) staining, and bisulfite genomic sequencing. No significant differences in spatial learning or phenotypic abnormality were found in adults derived from IVF (F1) and female and male line F2 and F3 generations. A borderline trend of hypomethylation was found in H19 DMR CpG island 3 in the female line-derived F3 generation (0.40±0.118, P=0.086). Methylation status in H19/Igf2 DMR island 1, Igf2 DMR, KvDMR, and Snrpn DMR displayed normal patterns. Methylation percentage did not differ significantly from that of adults conceived naturally, and the expression of the genes they regulated was not disturbed. Transgenerational integrity, such as behavior, morphology, histology, and DNA methylation status, was maintained in these generations, which indicates that exposure of female germ cells to hormonal stimulation and gamete manipulation might not affect the individuals and their descendents.
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References
Anway, M.D., Cupp, A.S., Uzumcu, M., Skinner, M.K., 2005. Epigenetic transgenerational actions of endocrine disruptors and male fertility. Science, 308(5727):1466–1469. [doi:10.1126/science.1108190]
Anway, M.D., Leathers, C., Skinner, M.K., 2006. Endocrine disruptor vinclozolin induced epigenetic transgenerational adult-onset disease. Endocrinology, 147(12):5515–5523. [doi:10.1210/en.2006-0640]
Bliek, J., Terhal, P., van den Bogaard, M.J., Maas, S., Hamel, B., Salieb-Beugelaar, G., Simon, M., Letteboer, T., van der Smagt, J., Kroes, H., et al., 2006. Hypomethylation of the H19 gene causes not only Silver-Russell syndrome (SRS) but also isolated asymmetry or an SRS-like phenotype. Am. J. Hum. Genet., 78(4):604–614. [doi:10.1086/502981]
Chan, T.L., Yuen, S.T., Kong, C.K., Chan, Y.W., Chan, A.S., Ng, W.F., Tsui, W.Y., Lo, M.W., Tam, W.Y., Li, V.S., et al., 2006. Heritable germline epimutation of MSH2 in a family with hereditary nonpolyposis colorectal cancer. Nat. Genet., 38(10):1178–1183. [doi:10.1038/ng1866]
Chao, M.J., Ramagopalan, S.V., Herrera, B.M., Lincoln, M.R., Dyment, D.A., Sadovnick, A.D., Ebers, G.C., 2009. Epigenetics in multiple sclerosis susceptibility: difference in transgenerational risk localizes to the major histocompatibility complex. Hum. Mol. Genet., 18(2):261–266. [doi:10.1093/hmg/ddn353]
Chin, H.J., Wang, C.K., 2001. Utero-tubal transfer of mouse embryos. Genesis, 30(2):77–81. [doi:10.1002/gene.1036]
Cox, G.F., Bürger, J., Lip, V., 2002. Intracytoplasmic sperm injection may increase the risk of imprinting defects. Am. J. Hum. Genet., 71(1):162–164. [doi:10.1086/341096]
Filipponi, D., Feil, R., 2009. Perturbation of genomic imprinting in oligozoospermia. Epigenetics, 4(1):27–30. [doi:10.4161/epi.4.1.7311]
Gluckman, P.D., Hanson, M.A., 2004. Living with the past: evolution, development and patterns of disease. Science, 305(5691):1733–1736. [doi:10.1126/science.1095292]
Hitchins, M.P., Wong, J.J., Suthers, G., Suter, C.M., Martin, D.I., Hawkins, N.J., Ward, R.L., 2007. Inheritance of a cancer-associated MLH1 germ-line epimutation. N. Engl. J. Med., 356(7):697–705. [doi:10.1056/NEJMoa064522]
Jirtle, R.L., Skinner, M.K., 2007. Environmental epigenomics and disease susceptibility. Nat. Rev. Genet., 8(4):253–262. [doi:10.1038/nrg2045]
Kobayashi, H., Sato, A., Otsu, E., Hiura, H., Tomatsu, C., Utsunomiya, T., Sasaki, H., Yaegashi, N., Arima, T., 2007. Aberrant DNA methylation of imprinted loci in sperm from oligospermic patients. Hum. Mol. Genet., 16(21):2542–2551. [doi:10.1093/hmg/ddm187]
Li, L.C., Dahiya, R., 2002. MethPrimer: designing primers for methylation PCRs. Bioinformatics, 18(11):1427–1431. [doi:10.1093/bioinformatics/18.11.1427]
Li, T., Vu, T.H., Ulaner, G.A., Littman, E., Ling, J.Q., Chen, H.L., Hu, J.F., Behr, B., Giudice, L., Hoffman, A.R., 2005. IVF results in de novo DNA methylation and histone methylation at an Igf2-H19 imprinting epigenetic switch. Mol. Hum. Reprod., 11(9):631–640. [doi:10.1093/molehr/gah230]
Lidegaard, O., Pinborg, A., Andersen, A.N., 2005. Imprinting diseases and IVF: Danish national IVF cohort study. Hum. Reprod., 20(4):950–954. [doi:10.1093/humrep/deh714]
Luuk, H., Plaas, M., Raud, S., Innos, J., Sütt, S., Lasner, H., Abramov, U., Kurrikoff, K., Kõks, S., Vasar, E., 2009. Wfs1-deficient mice display impaired behavioural adaptation in stressful environment. Behav. Brain Res., 198(2): 334–345. [doi:10.1016/j.bbr.2008.11.007]
Marques, C.J., Carvalho, F., Sousa, M., Barros, A., 2004. Genomic imprinting in disruptive spermatogenesis. Lancet, 363(9422):1700–1702. [doi:10.1016/S0140-6736(04)16256-9]
Middelburg, K.J., Heineman, M.J., Bos, A.F., Hadders-Algra, M., 2008. Neuromotor, cognitive, language and behavioural outcome in children born following IVF or ICSI—a systematic review. Hum. Reprod. Update, 14(3): 219–231. [doi:10.1093/humupd/dmn005]
Middelburg, K.J., Heineman, M.J., Bos, A.F., Pereboom, M., Fidler, V., Hadders-Algra, M., 2009. The Groningen ART cohort study: ovarian hyperstimulation and the in vitro procedure do not affect neurological outcome in infancy. Hum. Reprod., 24(12):3119–3126. [doi:10.1093/humrep/dep310]
Miller, C.A., Sweatt, J.D., 2007. Covalent modification of DNA regulates memory formation. Neuron, 53(6):857–869. [doi:10.1016/j.neuron.2007.02.022]
Morris, R., 1984. Developments of a water-maze procedure for studying spatial learning in the rat. J. Neurosci. Methods, 11(1):47–60. [doi:10.1016/0165-0270(84)90007-4]
Nadeau, J.H., 2009. Transgenerational genetic effects on phenotypic variation and disease risk. Hum. Mol. Genet., 18(R2):R202–R210. [doi:10.1093/hmg/ddp366]
Nilsson, E.E., Anway, M.D., Stanfield, J., Skinner, M.K., 2008. Transgenerational epigenetic effects of the endocrine disruptor vinclozolin on pregnancies and female adult onset disease. Reproduction, 135(5):713–721. [doi:10.1530/REP-07-0542]
Olson, C.K., Keppler-Noreuil, K.M., Romitti, P.A., Budelier, W.T., Ryan, G., Sparks, A.E., van Voorhis, B.J., 2005. In vitro fertilization is associated with an increase in major birth defects. Fertil. Steril., 84(5):1308–1315. [doi:10.1016/j.fertnstert.2005.03.086]
Ørstavik, K.H., Eiklid, K., van der Hagen, C.B., Spetalen, S., Kierulf, K., Skjeldal, O., Buiting, K., 2003. Another case of imprinting defect in a girl with Angelman syndrome who was conceived by intracytoplasmic semen injection. Am. J. Hum. Genet., 72(1):218–219. [doi:10.1086/346030]
Painter, R.C., Osmond, C., Gluckman, P., Hanson, M., Phillips, D.I., Roseboom, T.J., 2008. Transgenerational effects of prenatal exposure to the Dutch famine on neonatal adiposity and health in later life. BJOG, 115(10):1243–1249. [doi:10.1111/j.1471-0528.2008.01822.x]
Richards, E.J., 2006. Inherited epigenetic variation—revisiting soft inheritance. Nat. Rev. Genet., 7:395–401. [doi:10.1038/nrg1834]
Schneider, S., Kaufmann, W., Buesen, R., van Ravenzwaay, B., 2008. Vinclozolin—the lack of a transgenerational effect after oral maternal exposure during organogenesis. Reprod. Toxicol., 25(3):352–360. [doi:10.1016/j.reprotox.2008.04.001]
Skinner, M.K., 2008. What is an epigenetic transgenerational phenotype? F3 or F2. Reprod. Toxicol., 25(1):2–6. [doi:10.1016/j.reprotox.2007.09.001]
Stouder, C., Deutsch, S., Paoloni-Giacobino, A., 2009. Superovulation in mice alters the methylation pattern of imprinted genes in the sperm of the offspring. Reprod. Toxicol., 28(4):536–541. [doi:10.1016/j.reprotox.2009.06.009]
Whitelaw, N.C., Whitelaw, E., 2008. Transgenerational epigenetic inheritance in health and disease. Curr. Opin. Genet. Dev., 18(3):273–279. [doi:10.1016/j.gde.2008.07.001]
Xing, Y., Shi, S., Le, L., Lee, C.A., Silver-Morse, L., Li, W.X., 2007. Evidence for transgenerational transmission of epigenetic tumor susceptibility in Drosophila. PLoS Genet., 3(9):1598–1606. [doi:10.1371/journal.pgen.0030151]
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Project supported by the National Basic Research Program (973) of China (No. 2007CB948104), the National Natural Science Foundation of China (No. 81070532), and the Zhejiang Provincial Natural Science Foundation of China (No. Z207021)
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Li, L., Le, F., Wang, Ly. et al. Normal epigenetic inheritance in mice conceived by in vitro fertilization and embryo transfer. J. Zhejiang Univ. Sci. B 12, 796–804 (2011). https://doi.org/10.1631/jzus.B1000411
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DOI: https://doi.org/10.1631/jzus.B1000411
Key words
- Differentially methylated regions (DMRs)
- In vitro fertilization and embryo transfer (IVF-ET)
- Central nervous system (CNS)
- Neurobehavioral imprinting disorders
- Transgenerational epigenetic inheritance