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During early embryonic development, the retinoic acid signaling pathway coordinates with other signaling pathways to regulate body axis patterning and organogenesis. The production of retinoic acid requires two enzymatic reactions, the first of which is the oxidization of vitamin A (all-trans-retinol) to all-trans -retinal, mediated in part by the short-chain dehydrogenase/reductase. Through DNA microarrays, we have identified a gene in Xenopus laevis which shares a high sequence similarity to human short-chain dehydrogenase/reductase member 3. We therefore annotated the gene Xenopus short-chain dehydrogenase/reductase 3 (dhrs3). Expression of dhrs3 was detected by whole mount in situ hybridization in the dorsal blastopore lip and axial mesoderm region in gastrula embryos. During neurulation, dhrs3 transcripts were found in the notochord and neural ectoderm. Strong expression of dhrs3 was mainly detected in the brain, spinal cord and pronephros region in tailbud and tadpole stages. Temporal expression tested by RT-PCR indicated that dhrs3 was activated at the onset of gastrulation, and remained highly expressed at later stages of embryonic development. The distinct and highly regulated spatial and temporal expression of dhrs3 highlights the complexity of retinoic acid regulation.
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???displayArticle.pmcLink???PMC3418811 ???displayArticle.link???Int J Dev Biol ???displayArticle.grants???[+]
Fig. 2. Spatial expression pattern of dhrs3. Whole mount in situ hybridization was employed to examine the spatial expression of dhrs3 at different developmental stages as indicated. (A) Lateral view of a four-cell stage embryo. (B-F Dorsal view of early stage embryos. dhrs3 transcripts were localized in the dorsal blastopore lip and the ridge of neural fold. The expression of rdh10 in stage 11, 13 and 19 are shown in C'D'F' for comparison. (G-I) Frontal view of stage 13, 15, 19 embryos, which showed that dhrs3 expression was absent in the head, but present in the posterior edge of the up-folding neural tube. (J-L) Lateral view of stage 13, 15, 19 embryos, showing dhrs3 expression in ventral bilateral regions (arrow). (M-N Lateral views of tailbud and tadpole stage embryos, showing the expression pattern of dhrs3 and rdh10. The anterior region of a stage 35 embryo showing dhrs3 expression was magnified in (N). dhrs3 expression was detected in the forebrain, midbrain, pharynx, spinal cord and pronephros (arrow). In lateral views (J-N), anterior is to the left. Abbreviations: fb, forebrain; mb, midbrain; p, pharynx; sc, spinal cord; pn, pronephros.
Fig. 3. Transverse and longitudinal sections of embryos showing dhrs3 expression. Stage 11 embryo (A) was sectioned showing the expression
domain in dorsal blastopore lip and axial mesoderm (B). (C-D Transverse section from a stage 13 embryo (C), indicating the expression in notochord
and neuroectoderm. The higher magnification of framed region (D) is shown in (D. (E-G Longitudinal and transverse sections from a stage 15 embryo
(E) indicated dhrs3 was expressed in notochord and neuroectoderm. (H-N) Transverse sections of a stage 35 embryo (H) illustrating the expression in
the midbrain, lens, otic vesicles, ventralpharynx, gut, pronephros, dorsal spinal cord and notochord. Expression pattern of rdh10 is shown in (IM.
Abbreviations: bc, blastocoel; bpl, blastopore lip; ne, neuroepithelium; pm, paraxial mesoderm; nc, notochord; mb, midbrain; ov, otic vesicle; p, pharynx;
pn, pronephros; sc, spinal cord.
dhrs3 (dehydrogenase/reductase (SDR family) member 3) gene expression in Xenopus laevis embryos, NF stage 35, as assayed by in situ hybridization, lateral view, anteriorleft, dorsal up.
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