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Aberrant patterns of X chromosome inactivation in bovine clones

Nature Genetics volume 31, pages 216–220 (2002)Cite this article

Abstract

In mammals, epigenetic marks on the X chromosomes are involved in dosage compensation. Specifically, they are required for X chromosome inactivation (XCI), the random transcriptional silencing of one of the two X chromosomes in female cells during late blastocyst development1,2,3,4,5,6,7,8. During natural reproduction, both X chromosomes are active in the female zygote2. In somatic-cell cloning, however, the cloned embryos receive one active (Xa) and one inactive (Xi) X chromosome from the donor cells. Patterns of XCIhave been reported normal in cloned mice9, but have yet to be investigated in other species. We examined allele-specific expression of the X-linked monoamine oxidase type A (MAOA) gene and the expression of nine additional X-linked genes in nine cloned XX calves. We found aberrant expression patterns in nine of ten X-linked genes and hypomethylation of Xist in organs of deceased clones. Analysis of MAOA expression in bovine placentae from natural reproduction revealed imprinted XCI with preferential inactivation of the paternal X chromosome. In contrast, we found random XCI in placentae of the deceased clones but completely skewed XCI in that of live clones. Thus, incomplete nuclear reprogramming may generate abnormal epigenetic marks on the X chromosomes of cloned cattle, affecting both random and imprinted XCI.

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Figure 1: Establishment of assays for detection of MAOA allele–specific expression and XCI patterns in the donor animal.
Figure 2: Analysis of MAOA and Xist expression in major organs of a normal animal derived from natural reproduction and deceased clones.
Figure 3: Methylation patterns of bovine Xist in the hearts of a normal control and deceased clones E2 and I.
Figure 4: Allele-specific expression of MAOA and expression of Xist in bovine placentae.
Figure 5: Analysis of Xist expression and allele-specific expression of MAOA in live clones.

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Acknowledgements

The authors are indebted to H. Lewin and M. Band for providing primers for the bovine X-linked genes, to B. Jeong for initiating cell cultures, to R. Foote and M. Julian for critical reading of the manuscript and to J. Xu, B. Enright, S. Chaubal and J. Slisz for sample collection. This project was supported by grants from the National Institutes of Health, United States Department of Agriculture and Connecticut Innovations (to X.Y. and X.T.) and from Conselho Nacional ed Desenvolvimento Científico e Tecnológico and Fundação de Amparo à Pesquisa do Estado de São Paulo (to L.V.P.). This manuscript is a scientific contribution of the Storrs Agricultural Experiment Station at the University of Connecticut.

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  1. Department of Animal Science and Center for Regenerative Biology, University of Connecticut, Storrs, 06269, Connecticut, USA

    Fei Xue, X Cindy Tian, Fuliang Du, Chikara Kubota, Maneesh Taneja, Andras Dinnyes, Yunping Dai & Xiangzhong Yang

  2. Kagoshima Prefectural Cattle Breeding and Development Institute, Kagoshima, Japan

    Chikara Kubota

  3. Department of Environmental and Population Health, Tufts University School of Veterinary Medicine, South Woodstock, Connecticut, USA

    Howard Levine

  4. Departamento de Biologia, Centro de Estudos do Genoma Humano, Instituto de Biociências, Universidade de São Paulo, São Paulo, Brazil

    Lygia V. Pereira

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Xue, F., Tian, X., Du, F. et al. Aberrant patterns of X chromosome inactivation in bovine clones. Nat Genet 31, 216–220 (2002). https://doi.org/10.1038/ng900

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