We have cloned a novel retinoic acid (RA) catabolizing enzyme, Cyp26C1, in the chick and describe here its distribution during early stages of chick embryogenesis. It is expressed from stage 4 in the presumptive anterior (cephalic) mesoderm, in a subset of cephalic neural crest cells, the ventral otic vesicle, mesenchyme adjacent to the otic vesicle, the branchial pouches and grooves, a part of the neural retina, and the anterior telencephalon, and shows a dynamic expression in the hindbrain rhombomeres and neuronal populations within them. By examining the distribution of Cyp26C1 in the RA-free quail embryo, we can determine which of these expression domains is dependent on RA, and it is only the rhombomeric sites that do not appear, suggesting a role for RA in this location. The most striking domain of Cyp26C1 distribution is in the anterior cephalic mesoderm, which is adjacent to the domain of Raldh2 in the trunk mesoderm, but separated from it by a gap dorsal to which the posterior hindbrain will develop. We suggest that a gradient of RA within the mesoderm generated by Raldh2 and catabolized by Cyp26C1 could be responsible for patterning the hindbrain. We have compared this distribution of Cyp26C1 with that of Cyp26A1 and Cyp26B1 in the chick and shown that they generally occupy nonoverlapping sites of expression in the embryo, and as a result, we suggest individual roles for each of the Cyp enzymes in the developing embryo. Developmental Dynamics 230:509–517, 2004. © 2004 Wiley-Liss, Inc.