The retinoic acid metabolising gene, CYP26B1, patterns the cartilaginous cranial neural crest in zebrafish (original) (raw)
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Mechanisms of Development, 1997
Treatments of zebrafish embryos with retinoic acid (RA), a substance known to cause abnormal craniofacial cartilage development in other vertebrates, result in dose-and stage-dependent losses of dlx homeobox gene expression in several regions of the embryo. Dlx expression in neural crest cells migrating from the hindbrain and in the visceral arch primordia is particularly sensitive to RA treatment. The strongest effects are observed when RA is administered prior to or during crest cell migration but effects can also be observed if RA is applied when the cells have entered the primordia of the arches. Losses of dlx expression correlate either with the loss of cartilage elements originating from hindbrain neural crest cells or with abnormal morphology of these elements. Cartilage elements that originate from midbrain neural crest cells, which do not express dlx genes, are less affected. Taken together with the observation that the normal patterns of visceral arch dlx expression just prior to cartilage condensation resemble the morphology of the cartilage elements that are about to differentiate, our results suggest that dlx genes are an important part of a multi-step process in the development of a subset of craniofacial cartilage elements. 0 1997 Elsevier Science Ireland Ltd. All rights reserved
Feedback mechanisms regulate retinoic acid production and degradation in the zebrafish embryo
Mechanisms of Development, 2004
Retinoic acid (RA) signaling in vertebrate embryos occurs in a distinct physical and temporal pattern. Regulating this spatial distribution is crucial to the development of the embryo, as RA in excess or in inappropriate tissues is teratogenic. In order to understand how RA availability is determined in zebrafish we have investigated the expression of cyp26a1, an enzyme that inactivates RA, and its relationship to raldh2, one of the enzymes that produce RA from retinal. cyp26a1 expression follows three phases: in presumptive anterior neurectoderm and in a circumblastoporal ring during gastrulation, in the tailbud throughout somitogenesis, and in multiple specific tissue types beginning at mid-somitogenesis and continuing through 48 h postfertilization (hpf). This expression was either adjacent or opposite to those tissues expressing raldh2. We then investigated how RA production might regulate these relationships. Endogenous RA produced by raldhs did not play a role in setting cyp26a1 expression in most tissues. However, exogenous RA regulates expression of both enzymes. cyp26a1 is up regulated in the embryo in a time, concentration, and tissue-dependent manner. Conversely, raldh2 expression is reduced with RA treatment. Tests of the raldh2 promoter in cell transfections proved that RA directly represses its activity. These data demonstrate that the feedback mechanisms regulating production and degradation of RA must be considered in any experiments altering levels of RA in the developing vertebrate embryo.
Development, 2008
Skeletal syndromes are among the most common birth defects. Vertebrate skeletogenesis involves two major cell types: cartilage-forming chondrocytes and bone-forming osteoblasts. In vitro, both are under the control of retinoic acid (RA), but its exact in vivo effects remained elusive. Here, based on the positional cloning of the dolphin mutation, we have studied the role of the RA-oxidizing enzyme Cyp26b1 during cartilage and bone development in zebrafish. cyp26b1 is expressed in condensing chondrocytes as well as in osteoblasts and their precursors. cyp26b1 mutants and RA-treated wild-type fish display a reduction in midline cartilage and the hyperossification of facial and axial bones, leading to fusions of vertebral primordia, a defect not previously described in the context of RA signaling. Fusions of cervical vertebrae were also obtained by treating mouse fetuses with the specific Cyp26 inhibitor R115866. Together with data on the expression of osteoblast markers, our results i...
Retinoic acid is necessary for development of the ventral retina in zebrafish
Proceedings of the National Academy of Sciences, 1994
In the embryonic zebrafish retina, as in other vertebrates, retinoic acid is synthesized from retinaldehyde by two different dehydrogenases, one localized dorsally, the other primarily ventrally. Early in eye development only the ventral enzyme is present. Citral competitively inhibits the ventral enzyme in vitro and decreases the production of retinoic acid in the ventral retina in vivo. Treatment of neurula-stage zebrafish embryos with citral during the formation of the eye primordia results in eyes lacking a ventral retina. This defect can be partially rescued by retinoic acid. The results demonstrate that synthesis of retinoic acid can be selectively inhibited in vivo and suggest that retinoic acid is necessary for the proper development of the ventral retina.
Retinoic acid receptors' expression and function during zebrafish early development.
Retinoic acid (RA) regulates many developmental processes through its binding to two types of nuclear receptors, the retinoic acid receptor (RAR), and the retinoid-X receptor (RXR), which preferentially binds to the 9-cis isomer. Here we analyzed the RAR/RXR regulatory system during the first 5 days of development of zebrafish. Analysis of the relative transcript abundances for the four RAR and the six RXR zebrafish genes present in the zebrafish genome indicates a transition from maternal to embryonic transcripts during the first 24 h post fertilization. These changes did not affect the response to exogenous RA of the known RAR-responsive genes cyp26a1, dhrs3a, hoxb1b, hoxb5a, and hoxb5b. At the transcriptomic level, RA treatment elicited a negative feedback of genes involved in the endogenous RA synthesis and reduced levels of transcripts related to organ and anatomic development. These effects occurred at concentrations at which no morphological changes were observed. Data analysis suggests that exposure to exogenous RA results in an advance of the developing program, activating genes that should remain silent until later developmental stages and inhibiting expression of development-related genes. We conclude that zebrafish embryos are particularly sensitive to potential disruptors of the RAR/RXR regulatory system.
ZebRA: An overview of retinoic acid signalling during zebrafish development
2014
Retinoic Acid (RA), the main active vitamin A derivative,is crucial for embryo development,regulating cellular processes, embryo patterning and organogenesis. Many studies performed in mammalian or avian models have successfully undertakenthe investigation of the role played by RA during embryogenesis. However, since the early 1980's,the zebrafish (Daniorerio) has emerged as a powerful developmental model to study the in vivo role of RA during embryogenesis. Unlike other vertebrate models, zebrafish embryogenesis is external, allowing the observation of the translucent embryo from the earliest steps, but also providing an easily accessible system for pharmacological treatment or genetic approaches. Therefore, zebrafish research largely participates in deciphering the role of RA during development and this review aims at illustrating different concepts of RA signalling based on the research performed on zebrafish. Indeed, RA action relies on a multitude of cross-talkwith other signalling pathways and requires a coordinated, dynamic and fine-regulation of its level and activity in both temporal and spatial dimensions. This review also highlights major advances that have been discovered using zebrafish such as the observation of the RA gradient in vivofor the first time, the effects of RA signalling in brain patterning,its role in establishing left right asymmetry and its effects on the development of a variety of organs and tissues includingthe heart, blood, bone and fat. This review demonstrates thatthe zebrafish is a convenient and powerful model to study retinoic acid signalling during vertebrate embryogenesis.
Development (Cambridge, England), 2002
A number of studies have suggested that retinoic acid (RA) is an important signal for patterning the hindbrain, the branchial arches and the limb bud. Retinoic acid is thought to act on the posterior hindbrain and the limb buds at somitogenesis stages in chick and mouse embryos. Here we report a much earlier requirement for RA signalling during pre-segmentation stages for proper development of these structures in zebrafish. We present evidence that a RA signal is necessary during pre-segmentation stages for proper expression of the spinal cord markers hoxb5a and hoxb6b, suggesting an influence of RA on anteroposterior patterning of the neural plate posterior to the hindbrain. We report the identification and expression pattern of the zebrafish retinaldehyde dehydrogenase2 (raldh2/aldh1a2) gene. Raldh2 synthesises retinoic acid (RA) from its immediate precursor retinal. It is expressed in a highly ordered spatial and temporal fashion during gastrulation in the involuting mesoderm and...
Complex Regulation of cyp26a1 Creates a Robust Retinoic Acid Gradient in the Zebrafish Embryo
PLoS Biology, 2007
Positional identities along the anterior-posterior axis of the vertebrate nervous system are assigned during gastrulation by multiple posteriorizing signals, including retinoic acid (RA), fibroblast growth factors (Fgfs), and Wnts. Experimental evidence has suggested that RA, which is produced in paraxial mesoderm posterior to the hindbrain by aldehyde dehydrogenase 1a2 (aldh1a2/raldh2), forms a posterior-to-anterior gradient across the hindbrain field, and provides the positional information that specifies the locations and fates of rhombomeres. Recently, alternative models have been proposed in which RA plays only a permissive role, signaling wherever it is not degraded. Here we use a combination of experimental and modeling tools to address the role of RA in providing long-range positional cues in the zebrafish hindbrain. Using cell transplantation and implantation of RA-coated beads into RA-deficient zebrafish embryos, we demonstrate that RA can directly convey graded positional information over long distances. We also show that expression of Cyp26a1, the major RA-degrading enzyme during gastrulation, is under complex feedback and feedforward control by RA and Fgf signaling. The predicted consequence of such control is that RA gradients will be both robust to fluctuations in RA synthesis and adaptive to changes in embryo length during gastrulation. Such control also provides an explanation for the fact that loss of an endogenous RA gradient can be compensated for by RA that is provided in a spatially uniform manner.
Effects of retinoic acid exposure during zebrafish retinogenesis
Neurotoxicology and Teratology, 2013
Retinoic acid (RA) is an important morphogen involved in retinal development. Perturbations in its levels cause retinal malformations such as microphthalmia. However, the cellular changes in the retina that lead to this phenotype are little known. We have used the zebrafish to analyse the effects of systemic high RA levels on retinogenesis. For this purpose we exposed zebrafish embryos to 0.1 μM or 1 μM RA from 24 to 48 h post-fertilisation (hpf), the period which corresponds to the time of retinal neurogenesis and initial retinal cell differentiation. We did not find severe alterations in 0.1 μM RA treated animals, but the exposure to 1 μM RA significantly reduced retinal size upon treatment, and this microphthalmia persisted through larval development. We monitored histology and cell death and quantified both the proliferation rate and cell differentiation from 48 hpf onwards, focusing on the retina and optic nerve of normal and 1 μM treated animals. Retinal lamination and initial neurogenesis are not affected by RA exposure, but we found widespread apoptosis after RA treatment that could be the main cause of microphthalmia. Proliferating cells increased their number at 3 days post-fertilisation (dpf) but decreased significantly at 5 dpf maintaining the microphthalmic phenotype. Retinal cell differentiation was affected; some cell markers do not reach normal levels at larval stages and some cell types present an increased number compared to those of control animals. We also found the presence of young axons growing ectopically within the retina. Moreover although the optic axons leave the retina and form the optic chiasm they do not reach the optic tectum. The alterations observed in treated animals become more severe as larvae develop.
Genes & Development, 2001
Retinoic acid (RA), a derivative of vitamin A, plays a pivotal role in vertebrate development. The level of RA may be determined by the balance between its synthesis and degradation. We have examined the role of CYP26, a P450 enzyme that may degrade RA, by generating mutant mice that lack CYP26. CYP26−/− mice exhibited anomalies, including caudal agenesis, similar to those induced by administration of excess RA. The concentration of endogenous RA, as revealed by marker gene activity, was markedly increased in the tailbud of the mutant animals, in which CYP26 is normally expressed. Expression of T (Brachyury) andWnt3a in the tailbud was down-regulated inCYP26−/− mice, which may underlie the caudal truncation. The lack of CYP26 also resulted in homeotic transformation of vertebrae as well as in misspecification of the rostral hindbrain associated with anterior expansion of RA-positive domains. These results suggest that local degradation of RA by CYP26 is required for establishing an ...