cyp26a1 encodes cytochrome P450 26A1, an endoplasmic-reticulum retinoic-acid hydroxylase that catabolizes all-trans retinoic acid and thereby shapes retinoic-acid signaling and developmental patterning during zebrafish development.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0034653 retinoic acid catabolic process | IBA GO_REF:0000033 | ACCEPT | Summary: retinoic acid catabolic process (GO:0034653) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Zebrafish Cyp26a1 catalyzes oxidative metabolism of atRA, generating metabolites dominated by **4-hydroxy-RA (4-OH-RA)** and **4-oxo-RA (4-oxo-RA)** in microsome assays from transfected cells |
| GO:0007417 central nervous system development | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: central nervous system development (GO:0007417) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites |
| GO:0004497 monooxygenase activity | IEA GO_REF:0000002 | KEEP AS NON CORE | Summary: monooxygenase activity (GO:0004497) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md CYP26 enzymes are described as **membrane-anchored microsomal (endoplasmic reticulum, ER) cytochrome P450s** with a heme center |
| GO:0005506 iron ion binding | IEA GO_REF:0000002 | KEEP AS NON CORE | Summary: iron ion binding (GO:0005506) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md each catalytic cycle requires electrons supplied from **NADPH via cytochrome P450 oxidoreductase (POR)**, which uses **FAD and FMN** cofactors |
| GO:0005789 endoplasmic reticulum membrane | IEA GO_REF:0000044 | ACCEPT | Summary: endoplasmic reticulum membrane (GO:0005789) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Cyp26a1 is best supported as a **microsomal/ER membrane-anchored cytochrome P450**, i.e., positioned to access intracellular RA pools and regulate RA available for nuclear receptor signaling |
| GO:0008401 retinoic acid 4-hydroxylase activity | IEA GO_REF:0000117 | ACCEPT | Summary: retinoic acid 4-hydroxylase activity (GO:0008401) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Reviews of CYP26A1 metabolism emphasize **4-hydroxylation as the primary transformation** for CYP26A1 (and CYP26B1) |
| GO:0016705 oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen | IEA GO_REF:0000002 | KEEP AS NON CORE | Summary: oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen (GO:0016705) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites |
| GO:0020037 heme binding | IEA GO_REF:0000002 | KEEP AS NON CORE | Summary: heme binding (GO:0020037) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md CYP26 enzymes are described as **membrane-anchored microsomal (endoplasmic reticulum, ER) cytochrome P450s** with a heme center |
| GO:0034653 retinoic acid catabolic process | IEA GO_REF:0000117 | ACCEPT | Summary: retinoic acid catabolic process (GO:0034653) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md They convert RA into **more polar, generally less active metabolites**, supporting clearance and preventing ectopic signaling |
| GO:0062182 all-trans retinoic acid 4-hydrolase activity | IEA GO_REF:0000116 | ACCEPT | Summary: all-trans retinoic acid 4-hydrolase activity (GO:0062182) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Cell/microsome assays of zebrafish CYP26 family members show activity toward **RA isomers** (atRA, 9-cis RA, 13-cis RA) and **no detectable metabolism of retinol or retinal** under the tested conditions, supporting specialization for RA |
| GO:0005789 endoplasmic reticulum membrane | ISS GO_REF:0000024 | ACCEPT | Summary: endoplasmic reticulum membrane (GO:0005789) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: file:DANRE/cyp26a1/cyp26a1-uniprot.txt A cytochrome P450 monooxygenase involved in the metabolism of all-trans retinoic acid PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Experimental enzymology for zebrafish cyp26a1 was performed using **microsomes isolated from transfected cells**, which is consistent with ER-derived membrane localization |
| GO:0062182 all-trans retinoic acid 4-hydrolase activity | EXP PMID:8939936 Identification of the retinoic acid-inducible all-trans-reti... | ACCEPT | Summary: all-trans retinoic acid 4-hydrolase activity (GO:0062182) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Zebrafish Cyp26a1 catalyzes oxidative metabolism of atRA, generating metabolites dominated by **4-hydroxy-RA (4-OH-RA)** and **4-oxo-RA (4-oxo-RA)** in microsome assays from transfected cells |
| GO:0003151 outflow tract morphogenesis | IGI PMID:27893754 Cyp26 Enzymes Facilitate Second Heart Field Progenitor Addit... | KEEP AS NON CORE | Summary: outflow tract morphogenesis (GO:0003151) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:27893754 zebrafish embryos deficient for Cyp26a1 and Cyp26c1 enzymes, which promote RA degradation, have OFT defects |
| GO:0034672 anterior/posterior pattern specification involved in pronephros development | IMP PMID:27406002 BMP and retinoic acid regulate anterior-posterior patterning... | KEEP AS NON CORE | Summary: anterior/posterior pattern specification involved in pronephros development (GO:0034672) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:27406002 posterior kidney progenitors are protected ventrally by the RA-catabolizing enzyme Cyp26a1 file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md cyp26a1 is expressed early in presumptive anterior neural ectoderm, and later in forebrain, midbrain, anterior hindbrain, and tailbud territories, contributing to establishment of anterior RA-depleted domains opposing posterior RA synthesis |
| GO:0007507 heart development | IMP PMID:23990796 Depletion of retinoic acid receptors initiates a novel posit... | KEEP AS NON CORE | Summary: heart development (GO:0007507) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:23990796 Cyp26a1, an enzyme that facilitates degradation of RA |
| GO:0007507 heart development | IGI PMID:23990796 Depletion of retinoic acid receptors initiates a novel posit... | KEEP AS NON CORE | Summary: heart development (GO:0007507) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:23990796 Cyp26a1, an enzyme that facilitates degradation of RA |
| GO:0030917 midbrain-hindbrain boundary development | IGI PMID:23990796 Depletion of retinoic acid receptors initiates a novel posit... | KEEP AS NON CORE | Summary: midbrain-hindbrain boundary development (GO:0030917) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:23990796 Cyp26a1, an enzyme that facilitates degradation of RA |
| GO:0001944 vasculature development | IGI PMID:24667328 Cyp26 enzymes are required to balance the cardiac and vascul... | KEEP AS NON CORE | Summary: vasculature development (GO:0001944) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:24667328 Cyp26 enzymes largely act cell non-autonomously to promote appropriate cardiovascular development |
| GO:0055014 atrial cardiac muscle cell development | IGI PMID:24667328 Cyp26 enzymes are required to balance the cardiac and vascul... | KEEP AS NON CORE | Summary: atrial cardiac muscle cell development (GO:0055014) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:24667328 Cyp26 enzymes largely act cell non-autonomously to promote appropriate cardiovascular development |
| GO:0048384 retinoic acid receptor signaling pathway | IMP PMID:23975936 Retinoic acid-dependent regulation of miR-19 expression elic... | KEEP AS NON CORE | Summary: retinoic acid receptor signaling pathway (GO:0048384) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:23975936 A reporter assay confirmed that cyp26a1 is a bona fide target of miR-19 in vivo file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md a core component of **RA homeostasis** that sculpts spatial RA signaling territories in early embryos |
| GO:0003131 mesodermal-endodermal cell signaling | IMP PMID:19416885 Cyp26 enzymes function in endoderm to regulate pancreatic fi... | KEEP AS NON CORE | Summary: mesodermal-endodermal cell signaling (GO:0003131) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:19416885 the RA-degrading Cyp26 enzymes play a critical role in defining the normal anterior limit of the pancreatic field |
| GO:0031016 pancreas development | IMP PMID:19416885 Cyp26 enzymes function in endoderm to regulate pancreatic fi... | KEEP AS NON CORE | Summary: pancreas development (GO:0031016) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:19416885 the RA-degrading Cyp26 enzymes play a critical role in defining the normal anterior limit of the pancreatic field |
| GO:0031016 pancreas development | IGI PMID:19416885 Cyp26 enzymes function in endoderm to regulate pancreatic fi... | KEEP AS NON CORE | Summary: pancreas development (GO:0031016) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:19416885 the RA-degrading Cyp26 enzymes play a critical role in defining the normal anterior limit of the pancreatic field |
| GO:0034653 retinoic acid catabolic process | IMP PMID:19416885 Cyp26 enzymes function in endoderm to regulate pancreatic fi... | ACCEPT | Summary: retinoic acid catabolic process (GO:0034653) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: PMID:19416885 the RA-degrading Cyp26 enzymes play a critical role in defining the normal anterior limit of the pancreatic field |
| GO:0021797 forebrain anterior/posterior pattern specification | IGI PMID:17998248 Zebrafish model of holoprosencephaly demonstrates a key role... | KEEP AS NON CORE | Summary: forebrain anterior/posterior pattern specification (GO:0021797) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:17998248 The loss of the forebrain-specific RA-degrading enzyme cyp26a1 causes a forebrain phenotype that mimics tgif morphants |
| GO:0048854 brain morphogenesis | IMP PMID:17998248 Zebrafish model of holoprosencephaly demonstrates a key role... | KEEP AS NON CORE | Summary: brain morphogenesis (GO:0048854) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:17998248 The loss of the forebrain-specific RA-degrading enzyme cyp26a1 causes a forebrain phenotype that mimics tgif morphants |
| GO:0071299 cellular response to vitamin A | IDA PMID:17253779 Specificity of zebrafish retinol saturase: formation of all-... | KEEP AS NON CORE | Summary: cellular response to vitamin A (GO:0071299) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:17253779 all-trans-13,14-dihydroretinol is transiently oxidized to all-trans-13,14-dihydroretinoic acid before being oxidized further by Cyp26 enzymes |
| GO:0001756 somitogenesis | IMP PMID:17098223 Coordination of symmetric cyclic gene expression during somi... | KEEP AS NON CORE | Summary: somitogenesis (GO:0001756) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:17098223 expression of the RA-degrading enzyme cyp26a1 in the tailbud was controlled by Su(H) activity file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md microinjection of cyp26a1 mRNA reduces endogenous RA activity and yields phenotypes resembling reduced-RA conditions, consistent with a role in RA clearance |
| GO:0021661 rhombomere 4 morphogenesis | IMP PMID:17164423 Cyp26 enzymes generate the retinoic acid response pattern ne... | KEEP AS NON CORE | Summary: rhombomere 4 morphogenesis (GO:0021661) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:17164423 metabolize RA into polar derivatives, function redundantly to shape RA-dependent gene-expression domains during hindbrain development |
| GO:0021661 rhombomere 4 morphogenesis | IGI PMID:17164423 Cyp26 enzymes generate the retinoic acid response pattern ne... | KEEP AS NON CORE | Summary: rhombomere 4 morphogenesis (GO:0021661) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:17164423 metabolize RA into polar derivatives, function redundantly to shape RA-dependent gene-expression domains during hindbrain development |
| GO:0030902 hindbrain development | IGI PMID:17164423 Cyp26 enzymes generate the retinoic acid response pattern ne... | KEEP AS NON CORE | Summary: hindbrain development (GO:0030902) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:17164423 metabolize RA into polar derivatives, function redundantly to shape RA-dependent gene-expression domains during hindbrain development file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Hernandez et al. (2007; published Jan 2007; https://doi.org/10.1242/dev.02706) demonstrate that zebrafish orthologs of mammalian CYP26 genes (**cyp26a1, cyp26b1, cyp26c1**) act **redundantly** to shape the RA response pattern necessary for hindbrain development |
| GO:0042573 retinoic acid metabolic process | IMP PMID:17164423 Cyp26 enzymes generate the retinoic acid response pattern ne... | ACCEPT | Summary: retinoic acid metabolic process (GO:0042573) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: PMID:17164423 metabolize RA into polar derivatives, function redundantly to shape RA-dependent gene-expression domains during hindbrain development file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md They convert RA into **more polar, generally less active metabolites**, supporting clearance and preventing ectopic signaling |
| GO:0042574 retinal metabolic process | IMP PMID:17164423 Cyp26 enzymes generate the retinoic acid response pattern ne... | REMOVE | Summary: retinal metabolic process (GO:0042574) is not the appropriate direct annotation for cyp26a1. Reason: The synthesized evidence supports the specific core annotations reviewed separately; this broad or wrong-context annotation should be retired rather than treated as a core function. Supporting Evidence: PMID:17164423 metabolize RA into polar derivatives, function redundantly to shape RA-dependent gene-expression domains during hindbrain development |
| GO:0042573 retinoic acid metabolic process | IDA PMID:16455818 A novel cytochrome P450, zebrafish Cyp26D1, is involved in m... | ACCEPT | Summary: retinoic acid metabolic process (GO:0042573) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: PMID:16455818 Cyp26D1 expressed in 293T cells could metabolize all-trans RA, 9-cis RA, and 13-cis RA file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Cell/microsome assays of zebrafish CYP26 family members show activity toward **RA isomers** (atRA, 9-cis RA, 13-cis RA) and **no detectable metabolism of retinol or retinal** under the tested conditions, supporting specialization for RA |
| GO:0001568 blood vessel development | IMP PMID:15680360 Retinoic acid-metabolizing enzyme Cyp26a1 is essential for d... | KEEP AS NON CORE | Summary: blood vessel development (GO:0001568) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:15680360 the gene for the RA-degrading enzyme Cyp26a1 is mutated |
| GO:0030902 hindbrain development | IMP PMID:15680360 Retinoic acid-metabolizing enzyme Cyp26a1 is essential for d... | KEEP AS NON CORE | Summary: hindbrain development (GO:0030902) is retained as supported context for cyp26a1 but is not the primary/core function. Reason: This annotation is broad, inferred, or reflects downstream developmental/physiological context rather than the central molecular role. Supporting Evidence: PMID:15680360 the gene for the RA-degrading enzyme Cyp26a1 is mutated |
| GO:0008401 retinoic acid 4-hydroxylase activity | IDA PMID:8939936 Identification of the retinoic acid-inducible all-trans-reti... | ACCEPT | Summary: retinoic acid 4-hydroxylase activity (GO:0008401) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Zebrafish Cyp26a1 catalyzes oxidative metabolism of atRA, generating metabolites dominated by **4-hydroxy-RA (4-OH-RA)** and **4-oxo-RA (4-oxo-RA)** in microsome assays from transfected cells |
| GO:0042573 retinoic acid metabolic process | IDA PMID:8939936 Identification of the retinoic acid-inducible all-trans-reti... | ACCEPT | Summary: retinoic acid metabolic process (GO:0042573) is supported for cyp26a1. Reason: This annotation matches the synthesized core function or a directly supported core location/process for this gene. Supporting Evidence: PMID:8939936 all-trans-RA is rapidly metabolized to more polar metabolites file:DANRE/cyp26a1/cyp26a1-deep-research-falcon.md Reviews of CYP26A1 metabolism emphasize **4-hydroxylation as the primary transformation** for CYP26A1 (and CYP26B1), with additional products such as **18-hydroxy-RA** and more polar secondary metabolites |
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Download this section (compressed HTML)Q: To what extent do cyp26a1, cyp26b1, and cyp26c1 act redundantly versus in distinct spatial domains to shape the embryonic retinoic-acid gradient, and what is the unique non-redundant contribution of cyp26a1?
Q: Is cyp26a1 transcription controlled primarily by retinoic-acid-driven feedback (and post-transcriptionally by miR-19) in vivo, and how does this feedback set the dynamic range and robustness of retinoic-acid signaling?
Q: Does zebrafish Cyp26a1 act strictly as an all-trans retinoic acid 4-hydroxylase, or does it also generate 18-hydroxy and other polar metabolites that retain or lack signaling activity?
Experiment: Generate single and compound cyp26a1/cyp26b1/cyp26c1 CRISPR/Cas9 mutants and compare retinoic-acid reporter activity and hox/krox20 rhombomere boundary positioning to dissect each paralog's spatial contribution.
Hypothesis: cyp26a1 has a non-redundant role in clearing retinoic acid from anterior/hindbrain territories that is not fully compensated by cyp26b1 or cyp26c1.
Type: CRISPR/Cas9 genetics with retinoic-acid reporter imaging and in situ hybridization
Experiment: Express and purify recombinant Cyp26a1, incubate with all-trans, 9-cis, and 13-cis retinoic acid, and quantify the metabolite profile and kinetic parameters by LC-MS/MS to define substrate and regiochemical specificity.
Hypothesis: Recombinant zebrafish Cyp26a1 preferentially 4-hydroxylates all-trans retinoic acid rather than 9-cis or 13-cis isomers.
Type: in vitro cytochrome P450 enzyme assay with LC-MS/MS metabolite profiling
Experiment: Apply temporally controlled cyp26a1 loss-of-function and rescue, then quantify progenitor markers and retinoic-acid reporter activity in each tissue to test whether progenitor protection is a direct cyp26a1 function.
Hypothesis: Endogenous cyp26a1 is required to protect specific progenitor populations (posterior kidney, second heart field, pancreas) from retinoic-acid excess.
Type: conditional/temporal gene perturbation with marker quantification
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