Article | Published:

Somatic mosaicism in neuronal precursor cells mediated by L1 retrotransposition

Nature volume 435, pages 903910 (16 June 2005) | Download Citation

Subjects

Abstract

Revealing the mechanisms for neuronal somatic diversification remains a central challenge for understanding individual differences in brain organization and function. Here we show that an engineered human LINE-1 (for long interspersed nuclear element-1; also known as L1) element can retrotranspose in neuronal precursors derived from rat hippocampus neural stem cells. The resulting retrotransposition events can alter the expression of neuronal genes, which, in turn, can influence neuronal cell fate in vitro. We further show that retrotransposition of a human L1 in transgenic mice results in neuronal somatic mosaicism. The molecular mechanism of action is probably mediated through Sox2, because a decrease in Sox2 expression during the early stages of neuronal differentiation is correlated with increases in both L1 transcription and retrotransposition. Our data therefore indicate that neuronal genomes might not be static, but some might be mosaic because of de novo L1 retrotransposition events.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , , , & Neurogenesis in the adult brain: new strategies for central nervous system diseases. Annu. Rev. Pharmacol. Toxicol. 44, 399–421 (2004)

  2. 2.

    & Adult neurogenesis and neural stem cells of the central nervous system in mammals. J. Neurosci. Res. 69, 745–749 (2002)

  3. 3.

    Mammalian neural stem cells. Science 287, 1433–1438 (2000)

  4. 4.

    , & More hippocampal neurons in adult mice living in an enriched environment. Nature 386, 493–495 (1997)

  5. 5.

    , , & Running enhances neurogenesis, learning, and long-term potentiation in mice. Proc. Natl Acad. Sci. USA 96, 13427–13431 (1999)

  6. 6.

    et al. Initial sequencing and analysis of the human genome. Nature 409, 860–921 (2001)

  7. 7.

    et al. Genome sequence of the Brown Norway rat yields insights into mammalian evolution. Nature 428, 493–521 (2004)

  8. 8.

    et al. Initial sequencing and comparative analysis of the mouse genome. Nature 420, 520–562 (2002)

  9. 9.

    , & Defining the beginning and end of KpnI family segments. EMBO J. 3, 1753–1759 (1984)

  10. 10.

    & in Mobile DNA II (eds Craig, N., Craggie, R., Gellert, M. & Lambowitz, A.) 836–869 (ASM Press, Washington DC, 2002)

  11. 11.

    et al. Hot L1s account for the bulk of retrotransposition in the human population. Proc. Natl Acad. Sci. USA 100, 5280–5285 (2003)

  12. 12.

    , , & A novel active L1 retrotransposon subfamily in the mouse. Genome Res. 11, 1677–1685 (2001)

  13. 13.

    , , & Rapid amplification of a retrotransposon subfamily is evolving the mouse genome. Nature Genet. 20, 288–290 (1998)

  14. 14.

    Identification, characterization, and cell specificity of a human LINE-1 promoter. Mol. Cell. Biol. 10, 6718–6729 (1990)

  15. 15.

    , & A YY1-binding site is required for accurate human LINE-1 transcription initiation. Nucleic Acids Res. 32, 3846–3855 (2004)

  16. 16.

    , & Members of the SRY family regulate the human LINE retrotransposons. Nucleic Acids Res. 28, 411–415 (2000)

  17. 17.

    , , & An important role for RUNX3 in human L1 transcription and retrotransposition. Nucleic Acids Res. 31, 4929–4940 (2003)

  18. 18.

    From head to toes: the multiple facets of Sox proteins. Nucleic Acids Res. 27, 1409–1420 (1999)

  19. 19.

    et al. Sox2 regulatory sequences direct expression of a β-geo transgene to telencephalic neural stem cells and precursors of the mouse embryo, revealing regionalization of gene expression in CNS stem cells. Development 127, 2367–2382 (2000)

  20. 20.

    , & RE-1 silencing transcription factor (REST) regulates human synaptophysin gene transcription through an intronic sequence-specific DNA-binding site. Eur. J. Biochem. 270, 2–9 (2003)

  21. 21.

    & Epigenetic control of neural stem cell fate. Curr. Opin. Genet. Dev. 14, 461–469 (2004)

  22. 22.

    et al. High frequency retrotransposition in cultured mammalian cells. Cell 87, 917–927 (1996)

  23. 23.

    , , , & Determination of L1 retrotransposition kinetics in cultured cells. Nucleic Acids Res. 28, 1418–1423 (2000)

  24. 24.

    et al. Evidence consistent with human L1 retrotransposition in maternal meiosis I. Am. J. Hum. Genet. 71, 327–336 (2002)

  25. 25.

    , , , & Histone deacetylase inhibition-mediated neuronal differentiation of multipotent adult neural progenitor cells. Proc. Natl Acad. Sci. USA 101, 16659–16664 (2004)

  26. 26.

    et al. DNA repair mediated by endonuclease-independent LINE-1 retrotransposition. Nature Genet. 31, 159–165 (2002)

  27. 27.

    , & Exon shuffling by L1 retrotransposition. Science 283, 1530–1534 (1999)

  28. 28.

    , & Genomic deletions created upon LINE-1 retrotransposition. Cell 110, 315–325 (2002)

  29. 29.

    et al. Human l1 retrotransposition is associated with genetic instability in vivo. Cell 110, 327–338 (2002)

  30. 30.

    et al. Positional cloning of the gene for X-linked retinitis pigmentosa 2. Nature Genet. 19, 327–332 (1998)

  31. 31.

    & Tightly regulated, developmentally specific expression of the first open reading frame from LINE-1 during mouse embryogenesis. Proc. Natl Acad. Sci. USA 92, 1520–1524 (1995)

  32. 32.

    & Genetic and functional differences between multipotent neural and pluripotent embryonic stem cells. Proc. Natl Acad. Sci. USA 100(Suppl 1), 11866–11872 (2003)

  33. 33.

    et al. A mouse model of human L1 retrotransposition. Nature Genet. 32, 655–660 (2002)

  34. 34.

    , , & Tracking an embryonic L1 retrotransposition event. Proc. Natl Acad. Sci. USA 100, 1832–1837 (2003)

  35. 35.

    , & Transcriptional disruption by the L1 retrotransposon and implications for mammalian transcriptomes. Nature 429, 268–274 (2004)

  36. 36.

    et al. Disruption of the APC gene by a retrotransposal insertion of L1 sequence in a colon cancer. Cancer Res. 52, 643–645 (1992)

  37. 37.

    & RNA truncation by premature polyadenylation attenuates human mobile element activity. Nature Genet. 35, 363–366 (2003)

  38. 38.

    & Biology of mammalian L1 retrotransposons. Annu. Rev. Genet. 35, 501–538 (2001)

  39. 39.

    , & Isolation, characterization, and use of stem cells from the CNS. Annu. Rev. Neurosci. 18, 159–192 (1995)

  40. 40.

    et al. Survival and differentiation of adult neuronal progenitor cells transplanted to the adult brain. Proc. Natl Acad. Sci. USA 92, 11879–11883 (1995)

  41. 41.

    , , , & Software and methods for oligonucleotide and cDNA array data analysis. Genome Biol. 3, SOFTWARE0001.1–0001.9 (2002)

  42. 42.

    & Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. Proc. Natl Acad. Sci. USA 98, 31–36 (2001)

  43. 43.

    , , & Empirical characterization of the expression ratio noise structure in high-density oligonucleotide arrays. Genome Biol. 3, RESEARCH0018.1–0018.11 (2002)

  44. 44.

    , , & Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1994)

Download references

Acknowledgements

We thank M. L. Gage for editorial comments, H. Suh for Sox2-EGFP brain sections and embryo advice, P. Taupin for assistance during CCg experiments, and J. L. Garcia-Perez and R. Badge for critical comments on the manuscript. A.R.M. is a Pew Latin-America Fellow. V.T.C. was supported by grants from Lynn and Edward Streim and the Neuroplasticity of Aging Training Grant. J.V.M. was supported by grants from the National Institutes of Health and the W. M. Keck Foundation, and F.H.G. was supported by the Lookout Fund, The Christopher Reeve Paralysis Foundation, Max Planck Research Award Program, by the German Federal Ministry for Education, Science, Research and Technology and the National Institutes of Health: National Institute on Aging and National Institute of Neurological Disease and Stroke.Author Contributions A.R.M. is the leading author. He contributed to the concept, designed, performed the experiments and analysed the data, and wrote the manuscript. V.T.C. designed and performed the microarrays experiments. M.C.N.M. designed, performed and analysed the inverse PCR data and some tissue culture experiments and revised the manuscript. W.D. performed the transgenic experiment. J.V.M. contributed reagents, and performed data analyses and manuscript revision. F.H.G. is the senior author. He contributed to the concept, analysed the data, revised the manuscript and provided financial support.

Author information

Author notes

    • Vi T. Chu

    †Present address: Department of Cell Biology, Chemicon International, Inc., 28820 Single Oak Drive, Temecula, California 92590, USA

    • Alysson R. Muotri
    •  & Vi T. Chu

    *These authors contributed equally to this work

Affiliations

  1. Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA

    • Alysson R. Muotri
    • , Vi T. Chu
    • , Maria C. N. Marchetto
    • , Wei Deng
    •  & Fred H. Gage
  2. Department of Human Genetics and Internal Medicine, 1241 E. Catherine Street, University of Michigan Medical School, Ann Arbor, Michigan 48109-0618, USA

    • John V. Moran

Authors

  1. Search for Alysson R. Muotri in:

  2. Search for Vi T. Chu in:

  3. Search for Maria C. N. Marchetto in:

  4. Search for Wei Deng in:

  5. Search for John V. Moran in:

  6. Search for Fred H. Gage in:

Competing interests

Microarray data have been deposited in the GEO archive under accession number GSE2499, and the Cl22 L1 insertion sequence has been deposited in GenBank under accession number AY995186. Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Corresponding author

Correspondence to Fred H. Gage.

Supplementary information

Word documents

  1. 1.

    Supplementary Data

    Structures of L1 derived retrotransposition events in single cell clones from rat neural progenitor cells transfected with the L1-EGFP reporter construct (Word; 188 KB).

  2. 2.

    Supplementary Figure Legends

    Legends to accompany the below Supplementary Figures

  3. 3.

    Supplementary Methods

    Detailed, additional methods information to accompany the main manuscript.

  4. 4.

    Supplementary Notes

    Description of some neuronal genes targeted by L1 retrotransposition in neuronal precursor cells.

  5. 5.

    Supplementary Tables S1 and S2

    These tables resume the cloning survival after CCg-treatment and the transcripts obtained from the CCg-array experiment, respectively.

Videos

  1. 1.

    Supplementary Video

    This movie shows the neuronal differentiation of the Cl 22. The time lapse covers a period of 41h, during witch time the EGFP from the insertion site (Psd-93 gene) expression is turned on.

Image files

  1. 1.

    Supplementary Figure S1

    L1 transcripts are enriched in CCg-responsive cells.

  2. 2.

    Supplementary Figure S2

    Luciferase controls for the L1 5'UTR promoter analyses.

  3. 3.

    Supplementary Figure S3

    Adult NPCs derived from rat brain support L1 retrotransposition. FACS analysis.

  4. 4.

    Supplementary Figure S4

    Chromatin modification is associated with L1 insertion silencing.

  5. 5.

    Supplementary Figure S5

    Embryonic analysis of the L1-EGFP transgenic mice. Pregnant females were sacrificed at E10.5 and embryos were removed by micro-dissection.

  6. 6.

    Supplementary Figure S6

    Detection of L1 retrotransposition in other tissues of the L1-EGFP transgenic mice.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature03663

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.