title: New developmental evidence supports a homeotic frameshift of digit identity in the evolution of the bird wing.
abstract: BACKGROUND
  The homology of the digits in the bird wing is a high-profile controversy in developmental and evolutionary biology. The embryonic position of the digits cartilages with respect to the primary axis (ulnare and ulna) corresponds to 2, 3, 4, but comparative-evolutionary morphology supports 1, 2, 3. A homeotic frameshift of digit identity in evolution could explain how cells in embryonic positions 2, 3, 4 began developing morphologies 1, 2, 3. Another alternative is that no re-patterning of cell fates occurred, and the primary axis shifted its position by some other mechanism. In the wing, only the anterior digit lacks expression of HoxD10 and HoxD12, resembling digit 1 of other limbs, as predicted by 1, 2, 3. However, upon loss of digit 1 in evolution, the most anterior digit 2 could have lost their expression, deceitfully resembling a digit 1. To test this notion, we observed HoxD10 and HoxD12 in a limb where digit 2 is the most anterior digit: The rabbit foot. We also explored whether early inhibition of Shh signalling in the embryonic wing bud induces an experimental homeotic frameshift, or an experimental axis shift. We tested these hypotheses using DiI injections to study the fate of cells in these experimental wings.
  
  
  RESULTS
  We found strong transcription of HoxD10 and HoxD12 was present in the most anterior digit 2 of the rabbit foot. Thus, we found no evidence to question the use of HoxD expression as support for 1, 2, 3. When Shh signalling in early wing buds is inhibited, our fate maps demonstrate that an experimental homeotic frameshift is induced.
  
  
  CONCLUSIONS
  Along with comparative morphology, HoxD expression provides strong support for 1, 2, 3 identity of wing digits. As an explanation for the offset 2, 3, 4 embryological position, the homeotic frameshift hypothesis is consistent with known mechanisms of limb development, and further proven to be experimentally possible. In contrast, the underlying mechanisms and experimental plausibility of an axis shift remain unclear.
authors.affiliation: Laboratorio de Ontogenia y Filogenia. Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Ñuñoa, Santiago, Chile.
authors.forename: Miguel
authors.initials: M
authors.lastname: Salinas-Saavedra
authors.affiliation: Laboratorio de Ontogenia y Filogenia. Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Ñuñoa, Santiago, Chile.
authors.forename: Cristian
authors.initials: C
authors.lastname: Gonzalez-Cabrera
authors.affiliation: Laboratorio de Ontogenia y Filogenia. Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Ñuñoa, Santiago, Chile.
authors.forename: Luis
authors.initials: L
authors.lastname: Ossa-Fuentes
authors.affiliation: Laboratorio de Ontogenia y Filogenia. Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Ñuñoa, Santiago, Chile.
authors.forename: Joao F
authors.initials: JF
authors.lastname: Botelho
authors.affiliation: Laboratorio de Ontogenia y Filogenia. Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Ñuñoa, Santiago, Chile.
authors.forename: Macarena
authors.initials: M
authors.lastname: Ruiz-Flores
authors.affiliation: Laboratorio de Ontogenia y Filogenia. Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Ñuñoa, Santiago, Chile.
authors.forename: Alexander O
authors.initials: AO
authors.lastname: Vargas
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