Increased apoptosis and early embryonic lethality in mice nullizygous for the Huntington's disease gene homologue

Article metrics

Abstract

The expansion of GAG triplet repeats in the translated region of the human HD gene, encoding a protein (huntingtin) of unknown function, is a dominant mutation leading to manifestation of Huntington's disease. Targeted disruption of the homologous mouse gene (Hdh), to examine the normal role of huntingtin, shows that this protein is functionally indispensable, since nullizygous embryos become developmentally retarded and disorganized, and die between days 8.5 and 10.5 of gestation. Based on the observation that the level of the regionalized apoptotic cell death in the embryonic ectoderm, a layer expressing the Hdh gene, is much higher than normal in the null mutants, we propose that huntingtin is involved in processes counterbalancing the operation of an apoptotic pathway.

Access 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

    Harper, P.S. Huntington's Disease. Major Problems in Neurology, vol. 22, (W.B. Saunders, London, 1991).

  2. 2

    Albin, R.L. & Tagle, D.A. Genetics and molecular biology of Huntington's disease. Trends Neurosci. 18, 11–14 (1995).

  3. 3

    Gusella, J.F. & MacDonald, M.E. Huntington's disease. Sem. Cell Biol. 6, 21–28 (1995).

  4. 4

    Albin, R.L., Young, A.B. & Penney, J.B. The functional anatomy of basal ganglia disorders. Trends Neurosci. 12: 366–375 (1989).

  5. 5

    Portera-Cailliau, C., Hedreen, J.C., Price, D.L. & Koliatsos, V.E. Evidence for apoptotic cell death in Huntigton disease and excitotoxic animal models. J. Neurosci. 15, 3775–3787 (1995).

  6. 6

    Thomas, L.B., Gates, D.J., Richfield, E.K., O'Brien, T.F., Schweitzer, J.B. & Steindler, D.A. DNA end labeling (TUNEL) in Huntington's disease and other neuropathological conditions. Exp. Neurol. (in the press).

  7. 7

    Huntington's Disease Collaborative Research Group. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes. Cell 72, 971–983 (1993).

  8. 8

    Ambrose, C.M. et al. Structure and expression of the Huntington's disease gene: evidence against simple inactivation due to an expanded GAG repeat. Som. Cell molec. Genet. 20, 27–38 (1994).

  9. 9

    Richards, R.I. & Sutherland, G.R. Dynamic mutations: a new class of mutations causing human disease. Cell 70, 709–712 (1992).

  10. 10

    Sutherland, G.R. & Richards, R.I. Dynamic mutations on the move. J. med. Genet. 30, 978–981 (1993).

  11. 11

    Sharp, A. et al. Widespread expression of Huntington's disease gene (IT15) protein product. Neuron 14, 1065–1074 (1995).

  12. 12

    Trottier, Y. et al. Cellular localization of the Huntington's disease protein and discrimination of the normal and mutated form. Nature Genet. 10, 104–110 (1995).

  13. 13

    DiFiglia, M. et al. Huntingtin is a cytoplasmic protein associated with vesicles in human and rat brain neurons. Neuron 14, 1075–1081 (1995).

  14. 14

    Jou, Y-S. & Myers, R.M. Evidence from antibody studies that the GAG repeat in the Huntington disease gene is expressed in the protein. Hum. molec. Genet. 4, 465–469 (1995).

  15. 15

    Kremer, H.P.H. et al. Worldwide study of the Huntington's disease mutation: the sensitivity and specificity of repeated GAG sequences. New Engl. J. Med. 330, 1401–1406 (1994).

  16. 16

    Duyao, M. et al. Trinucleotide repeat length instability and age of onset in Huntington's disease. Nature Genet. 4, 387–392 (1993).

  17. 17

    Telenius, H. et al. Molecular analysis of juvenile Huntington disease: the major influence on (CAG)n repeat length is the sex of the affected parent. Hum. molec. Genet. 2, 1535–1540 (1993).

  18. 18

    Stine, O.C., Pleasant, N., Franz, M.L., Abbott, M.H., Folstein, S.E. and Ross, C.A. Correlation between the onset age of Huntington's disease and length of the trinucleotide repeat in IT-15. Hum. molec. Genet. 2, 1547–1549 (1993).

  19. 19

    Andrew, S.E., Goldberg, Y.P., Theilmann, J., Zeisler, J. & Hayden, M.R. A CCG repeat polymorphism adjacent to the GAG repeat in the Huntington disease gene: implications for diagnostic accuracy and predictive testing. Hum. Molec. Genet. 3, 65–67 (1994).

  20. 20

    Barren, L.H., Rae, A., Holloway, S., Brock, D.J.H. & Warner, J.P. A single allele from the polymorphic CCG rich sequence immediately 3′ to the unstable CAG trinucleotide in the IT15 cDNA shows almost complete disequilibrium with the Huntington's disease chromosome in the Scottish population. Hum. molec. Genet. 3: 173–175 (1994).

  21. 21

    Li, S. et al. Huntington's disease gene (IT15) is widely expressed in human and rat tissues. Neuron 11, 985–993 (1993).

  22. 22

    Strong, T.V. et al. Widespread expression of the human and rat Huntington's disease gene in brain and nonneural tissues. Nature Genet. 5, 259–265 (1993).

  23. 23

    Hoogeveen, A.T. et al. Characterization and localization of the Huntington disease gene product. Hum. molec. Genet. 2, 2069–2073 (1993).

  24. 24

    Ambrose, M.P. et al. Huntington's disease gene: regional and cellular expression in brain of normal and affected individuals. Ann. Neurol. 37, 218–230 (1995).

  25. 25

    Gerber, H.-P. et al. Transcriptional activation modulated by homopolymeric glutamine and proline stretches. Science 263, 808–811 (1994).

  26. 26

    Barnes, G.T. et al. Mouse Huntington's disease gene homolog (Hdh). Som. Cell molec. Genet. 20: 87–97 (1994).

  27. 27

    Nasir, J. et al. Targeted disruption of the Huntington's disease gene results in embryonic lethality and behavioral and morphological changes in heterozygotes. Cell 81, 811–823 (1995).

  28. 28

    Duyao, M.P. et al. Inactivation of the mouse Huntington's disease gene homolog Hdh. Science 269, 407–410 (1995).

  29. 29

    Poelmann, R.E. & Vermeij-Keers, C. Cell degeneration in the mouse embryo: a prerequisite for normal development. In Progress in Differentiation Research (ed. Müller-Béra, N.) 93–102 (North Holland Publ. Co., Amsterdam, 1976).

  30. 30

    Poelmann, R.E. Morphological changes in the ectoderm of early mouse embryos related to the patterns of cell division and cell degeneration. J. Anat. 124, 238–240 (1977).

  31. 31

    Poelmann, R.E. Differential mitosis and degeneration patterns in relation to the alterations in the shape of the embryonic ectoderm of early post-implantation mouse embryos. J. Embryol. exp. Morph. 55, 33–51 (1980).

  32. 32

    Abrams, J.M., White, K., Fessler, L.I. & Steller, H. Programmed cell death during Drosophila embryogenesis. Development 117, 29–43 (1993).

  33. 33

    Gao, X., Blackburn, M.R. & Knudsen, T.B. Activation of apoptosis in early mouse embryos by 2′-deoxyadenosine exposure. Teratology 49, 1–12 (1994).

  34. 34

    Gavrieli, Y., Sherman, Y. & Ben-Sasson, S.A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119, 493–501 (1992).

  35. 35

    Jollie, W.P., Development, morphology, and function of the yolk-sac placenta in laboratory rodents. Teratology 41, 361–381 (1990).

  36. 36

    Kadokawa, Y., Kato, Y. & Eguchi, G. Cell lineage analysis of the primitive and visceral endoderm of mouse embryos cultured in vitro. Cell Diff. 21, 69–76 (1987).

  37. 37

    Chen, W.S. et al. Disruption of the HNF-4 gene, expressed in visceral endoderm, leads to cell death in embryonic ectoderm and impaired gastrulation of mouse embryos. Genes Dev. 8, 2466–2477 (1994).

  38. 38

    Copp, A.J. Death before birth: clues from gene knockouts and mutations. Trends Genet. 11, 87–93 (1995).

  39. 39

    Lewis, S.E., Turchin, H.A. & Gluecksohn-Waelsch, S. The developmental analysis of an embryonic lethal (c6H) in the mouse. J. Embryol. exp. Morph. 36, 363–371 (1976).

  40. 40

    Niswander, L., Yee, D., Rinchik, E.M., Russell, L.B. & Magnuson, T. The albino deletion complex and early postimplantation survival in the mouse. Development 102, 45–53 (1988).

  41. 41

    Yamaguchi, T.P., Harpal, K., Henkemeyer, M. & Rossant, J. fgfr-1 is required for embryonic growth and mesodermal patterning during mouse gastrulation. Genes Dev. 8, 3032–3044 (1994).

  42. 42

    Deng, C-X., Wynshaw-Boris, A., Shen, M.M., Daugherty, C., Ornitz, D.M. & Leder, P. Murine FGFR-1 is required for early postimplantation growth and axial organization. Genes Dev. 8, 3045–3057 (1994).

  43. 43

    Spyropoulos, D.D. & Capecchi, M.R. Targeted disruption of the even-skipped gene, evxl, causes early postimplantation lethality of the mouse conceptus. Genes Dev. 8, 1949–1961 (1994).

  44. 44

    Martin, S.J., Green, D.R. & Cotter, T.G. Dicing with death: dissecting the components of the apoptosis machinery. Trends biochem. Sci. 19, 26–30 (1994).

  45. 45

    Bellamy, C.O.C., Malcomson, R.D.G., Harrison, D.J. & Wyllie, A.H. Cell death in health and disease: the biology and regulation of apoptosis. Sem. Cancer Biol. 6, 3–16, (1995).

  46. 46

    Snow, M.H.L. & Tam, P.P.L. Is compensatory growth a complicating factor in mouse teratology? Nature 279, 555–557 (1979).

  47. 47

    Snow, M.H.L. Growth and its control in early mammalian development. Br. Med. Bull. 37, 221–226 (1981).

  48. 48

    Snow, M.H.L. Control of embryonic growth rate and fetal size in mammals. In Human Growth: A Comprehensive Treatise. (eds Falkner, F. & Tanner, J.M.) vol.3 67–82 (Plenum Press, New York, 1986).

  49. 49

    Raff, M.C. Social controls on cell survival and cell death. Nature 365, 397–400 (1992).

  50. 50

    Rabizadeh, S. et al. Mutations associated with amyotrophic lateral sclerosis convert superoxide dismutase from an antiapoptotic gene to a proapoptotic gene: studies in yeast and neural cells. Proc. natn. Acad. Sci. U.S.A. 92, 3024–3028 (1995).

  51. 51

    Lin, B. et al. Sequence of the murine Huntington disease gene: evidence for conservation, alternate splicing and polymorphism in a triplet (CCG) repeat. Hum. molec. Genet. 3, 1541–1545 (1994).

  52. 52

    Mansour, S.L., Thomas, K.R. & Capecchi, M.R. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature 336, 348–352 (1988).

  53. 53

    Thomas, K.R. & Capecchi, M.R. Targeted disruption of the murine int-1 proto-oncogene resulting in servere abnormalities in midbrain and cerebellar development. Nature 346, 847–850 (1990).

  54. 54

    Liu, J.-P., Baker, J., Perkins, A.S., Robertson, E.J. & Efstratiadis, A. Mice carrying null mutations of the genes encoding insulin-like growth factor I (Igf-1) and type 1 IGF receptor (Igf1r). Cell 75, 59–72 (1993).

  55. 55

    Robertson, E.J. Embryo-derived stem cell lines. In Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, (ed. Robertson, E.J.) 71–112 (IRL Press, Oxford, 1987).

  56. 56

    Bradley, A. Production and analysis of chimeric mice. In Teratocarcinomas and Embryonic Stem Cells: A Practical Approach. (ed. Robertson, E.J.) 131–151, (IRL Press, Oxford, 1987).

  57. 57

    Hogan, B., Beddington, R., Costantini, F & Lacy E. Manipulating the Mouse Embryo: A Laboratory Manual. (Cold Spring Harbor Laboratory Press, New York, 1994).

  58. 58

    Schubert, E.L. et al. A method to isolate DNA from small archival tissue samples for p53 gene analysis. Hum. Mutat. 2, 123–126 (1993).

  59. 59

    Greer, C.E., Wheeler, C.M. & Manos, M.M. Sample preparation and PCR amplification from paraffin-embedded tissues. PCR Methods Appl. 3, S113–8122 (1994).

  60. 60

    Harland, R.M. In situ hybridization: an improved whole mount method for Xenopus embryos. In Methods in Cell Biology. (eds Kay, B. K. & Peng, H. J.) 36, 675–685 (Academic Press, New York, 1991).

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Zeitlin, S., Liu, J., Chapman, D. et al. Increased apoptosis and early embryonic lethality in mice nullizygous for the Huntington's disease gene homologue. Nat Genet 11, 155–163 (1995) doi:10.1038/ng1095-155

Download citation

Further reading