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A reevaluation of X-irradiation-induced phocomelia and proximodistal limb patterning

Nature volume 460, pages 400–404 (2009)Cite this article

Abstract

Phocomelia is a devastating, rare congenital limb malformation in which the long bones are shorter than normal, with the upper portion of the limb being most severely affected. In extreme cases, the hands or fingers are attached directly to the shoulder and the most proximal elements (those closest to the shoulder) are entirely missing. This disorder, previously known in both autosomal recessive and sporadic forms, showed a marked increase in incidence in the early 1960s due to the tragic toxicological effects of the drug thalidomide, which had been prescribed as a mild sedative1,2. This human birth defect is mimicked in developing chick limb buds exposed to X-irradiation3,4,5. Both X-irradiation5 and thalidomide-induced phocomelia5,6 have been interpreted as patterning defects in the context of the progress zone model, which states that a cell’s proximodistal identity is determined by the length of time spent in a distal limb region termed the ‘progress zone’7. Indeed, studies of X-irradiation-induced phocomelia have served as one of the two major experimental lines of evidence supporting the validity of the progress zone model. Here, using a combination of molecular analysis and lineage tracing in chick, we show that X-irradiation-induced phocomelia is fundamentally not a patterning defect, but rather results from a time-dependent loss of skeletal progenitors. Because skeletal condensation proceeds from the shoulder to fingers (in a proximal to distal direction), the proximal elements are differentially affected in limb buds exposed to radiation at early stages. This conclusion changes the framework for considering the effect of thalidomide and other forms of phocomelia, suggesting the possibility that the aetiology lies not in a defect in the patterning process, but rather in progenitor cell survival and differentiation. Moreover, molecular evidence that proximodistal patterning is unaffected after X-irradiation does not support the predictions of the progress zone model.

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Figure 1: X-irradiation of chick limbs causes phocomelia.
Figure 2: Expression analysis of irradiated limbs reveals defects in differentiation but not specification of the proximal segments.
Figure 3: Changes in cell death and proliferation do not explain the loss of proximal structures.
Figure 4: X-irradiation disrupts chondrogenesis and does not convert proximal cells to distal fates.

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Acknowledgements

We would like to thank J. G. Arozamena and D. Connor for help with the irradiation machine and members of the Tabin-Cepko and Ros laboratories for discussions and ideas. Work in the laboratory of C.J.T. is supported by a grant, R37 HD032443, from the NIH. J.L.G. is supported by grant F32HD057701 from the National Institute of Child Health and Human Development. I.D. and M.A.R. are supported by BFU2008-00397 from the Spanish Ministry of Science and Innovation.

Author Contributions J.L.G. performed the shielding experiments. I.D. performed the grafting experiments. J.L.G., I.D., C.J.T. and M.A.R. came up with ideas, analysed data and wrote the paper. C.J.T. and M.A.R. oversaw the research.

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Author notes

  1. Jenna L. Galloway and Irene Delgado: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA,

    Jenna L. Galloway & Clifford J. Tabin

  2. Departamento de Anatomia y Biologia Celular, Universidad de Cantabria,

    Irene Delgado

  3. Instituto de Biomedicina y Biotecnología de Cantabria (CSIC-UC-IDICAN), C/ Herrera Oria s/n, 39011 Santander, Spain,

    Maria A. Ros

Authors
  1. Jenna L. Galloway
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  2. Irene Delgado
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  3. Maria A. Ros
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  4. Clifford J. Tabin
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Corresponding author

Correspondence to Clifford J. Tabin.

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Galloway, J., Delgado, I., Ros, M. et al. A reevaluation of X-irradiation-induced phocomelia and proximodistal limb patterning. Nature 460, 400–404 (2009). https://doi.org/10.1038/nature08117

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