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Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), CNRS/INSERM/ULP/Collège de France, B.P. 163, 67404 ILLKIRCH Cedex, C.U. de STRASBOURG, France
*Author for correspondence (e-mail: marek{at}igbmc.u-strasbg.fr)
Accepted March 13, 2001
Mutants mice carrying targeted inactivations of both retinoic acid receptor (RAR) 
and RAR
(A/A mutants) were analyzed at different embryonic stages, in order to establish the timing of appearance of defects that we previously observed during the fetal period. We show that embryonic day (E)9.5 A/A embryos display severe malformations, similar to those already described in retinaldehyde dehydrogenase 2 null mutants. These malformations reflect early roles of retinoic acid signaling in axial rotation, segmentation and closure of the hindbrain; formation of otocysts, pharyngeal arches and forelimb buds; and in the closure of the primitive gut. The hindbrain of E8.5 A/A embryos shows a posterior expansion of rhombomere 3 and 4 (R3 and R4) markers, but fails to express kreisler, a normal marker of R5 and R6. This abnormal hindbrain phenotype is strikingly different from that of embryos lacking RAR and RARß (A/Aßmutants), in which we have previously shown that the territory corresponding to R5 and R6 is markedly enlarged. Administration of a pan-RAR antagonist at E8.0 to wild-type embryos cultured in vitro results in an A/Aß-like hindbrain phenotype, whereas an earlier treatment at E7.0 yields an A/A-like phenotype. Altogether, our data suggest that RAR and/or RAR transduce the RA signal that is required first to specify the prospective R5/R6 territory, whereas RARß is subsequently involved in setting up the caudal boundary of this territory.
Key words: Nuclear receptors, Rhombomeres, Hox genes, kreisler, Embryo culture, Mouse, Vitamin A
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