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Retinoids long have been implicated in limb development and their endogenous contributions to this process are finally being elucidated. Here we use an established model of retinoid depletion during specific gestational windows to investigate the role of endogenous retinoic acid (RA) in supporting limb outgrowth. Rat embryos were deprived of RA starting at days‐postcoitum (dpc) 3.0, 5.5, or 7.0 and harvested at the 35‐somite stage (dpc 12–12.5). Although embryos from all these windows possessed many characteristics of gestational retinoid deficiency (frontonasal hypoplasia, straight tail, reduced CRBPI and RARβ), their limb buds emerged with only modest size reductions. Molecular analysis of RA‐deficient limb buds revealed enhanced gli‐3 and reduced hoxd‐12, hoxd‐13, shh, and fgf‐4, while fgf‐8, en‐1, and wnt‐7a expression remained unaltered. Occasional posterior truncations were observed at low incidence in the longest deficiency window; otherwise, the deficiency window length had no discernable impact on the severity of these changes. At the 45‐somite stage, RA‐deficient limbs had additional losses of hoxd‐13 and fgf‐8, accompanied by a flattened AER, suggestive of an ultimate failure in limb bud outgrowth. Results could not confirm a function for endogenous retionids in limb initiation, but show they are required to maintain the signaling loops between the developing mesenchyme and AER that govern limb outgrowth after the initial emergence of limb bud. Dev Dyn 1999;216:469–480. ©1999 Wiley‐Liss, Inc.
Developmental Dynamics – Wiley
Published: Dec 1, 1999
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