3D expression patterns of cell cycle genes in the developing chick wing and comparison with expression patterns of genes implicated in digit specification

Sonic hedgehog (Shh) signalling controls integrated specification of digit pattern and growth in the chick wing but downstream gene networks remain to be unravelled. We analysed 3D expression patterns of genes encoding cell cycle regulators using Optical Projection Tomography. Hierarchical clustering of spatial matrices of gene expression revealed a dorsal layer of the wing bud, in which almost all genes were expressed, and that genes encoding positive cell cycle regulators had similar expression patterns while those of N-myc and CyclinD2 were distinct but closely related. We compared these patterns computationally with those of genes implicated in digit specification and Ptch1 , 50 genes in total. Nineteen genes have similar posterior expression to Ptch1 , including Hoxd13, Sall1, Hoxd11 , and Bmp2 , all likely Gli targets in mouse limb, and cell cycle genes, N-myc , CyclinD2 . We suggest that these genes contribute to a network integrating digit specification and growth in response to Shh.