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Key Event Title
Decreased all-trans retinoic acid (atRA) concentration
|Level of Biological Organization|
Key Event Components
Key Event Overview
AOPs Including This Key Event
|AOP Name||Role of event in AOP||Point of Contact||Author Status||OECD Status|
|Inhibition of ALDH1A leading to reduced fertility, female||KeyEvent||Cataia Ives (send email)||Under development: Not open for comment. Do not cite||Under Development|
|RALDH2 and cardiovascular developmental defects||KeyEvent||Arthur Author (send email)||Open for comment. Do not cite|
|All life stages||Moderate|
Key Event Description
All-trans retinoic acid (atRA) is the active form of vitamin A/all-trans retinol and is involved in regulating a large number of developmental processes (Bushue & Wan, 2010a; Ghyselinck & Duester, 2019). Although 9-cis RA and 13-cis RA are other metabolic derivatives of vitamin A, atRA is generally considered the primary active metabolite during development, mainly acting as a short-range paracrine signaling molecule (Cunningham & Duester, 2015). atRA exerts dose-dependent effects on morphogenesis, so disruption to atRA concentrations during development can lead to malformations in numerous tissues and organs. During development the spatiotemporal regulation of atRA concentrations in target tissues is tightly controlled by a balance of synthesis and degradation enzymes (Kedishvili, 2013).
Cellular atRA synthesis starts by oxidation of vitamin A to retinaldehyde (RAL) by retinol dehydrogenase-10 (RDH10). RAL is then irreversibly converted to atRA by RAL dehydrogenases (ALDH1A1, ALD1A2, or ALDH1A3). To maintain appropriate retinoid levels in tissues, RAL can be converted back to retinol by enzymatic reactions; further retinoid levels can be controlled by enzymatic degradation of atRA by the cytochrome P450 enzymes CYP26A1, CYP26B1, or CYP26C1, which are differentially expressed throughout the mammalian body (Isoherranen & Zhong, 2019; Shimozono et al, 2013). Inhibition/disruption of any of the enzymes of the atRA synthesis pathway, or increased expression of the atRA degradation enzymes can lead to decreased concentrations of atRA in target cells (Kedishvili, 2013).
The atRA functions as a ligand for the nuclear retinoic acid receptors (RARs), which form heterodimers with the retinoid X receptors (RXRs); the atRA:RAR:RXR complex then binds to retinoic acid response elements (RAREs) upstream of target genes, leading to activation or repression of gene expression in target cells (Chambon, 1996; le Maire et al, 2019). The type and number of RAR/RXRs differ between evolutionary distant animals, but functionally they are all involved in the regulation of development (Gutierrez-Mazariegos et al, 2014).
How It Is Measured or Detected
Direct measurements of atRA in serum (humans, animals) can be performed by various chromatographic methods (Gundersen, 2006), including high performance liquid chromatography (HPLC) or liquid chromatography-tandem mass spectrometry (LC-MS) (Morgenstern et al, 2021).
Indirect measurements in cells and animal models can be performed with reporter assays utilizing RAR-RXR-RARE or RXR-RXR-RARE promoter elements, which are activated by atRA, driving expression of reporter proteins. These reporter assays can detect the presence of atRA in tissues in a semi-quantitative manner. Examples include reporter mouse lines (Carlsen et al, 2021; Rossant et al, 1991; Solomin et al, 1998), reporter cell lines (Wagner et al, 1992) and transient transfection of constructs for in vitro cell-based assays (Chassot et al, 2020).
Domain of Applicability
The retinoid signaling system is highly conserved across animal species (Bushue & Wan, 2010b; Rhinn & Dollé, 2012). atRA acts as a ligand for the nuclear retinoic acid (RAR) receptors, which upon activation regulate gene transcription in target cells. The type and number of RARs differ between evolutionary distant animals, but functionally they are all involved in the regulation of development.
Evidence for Perturbation by Stressor
WIN18,446, a RALDH2 inhibitor, inhibits RA biosynthesis resulting in reduced ATRA in mice: e.g. around 50% reduction in liver and 90% reduction in testes (Arnold et al, 2015).
Ethanol is also catalyzed by the enzyme alcohol dehydrogenase (ADH) and is known to compete with retinol for ADH-catalyzed oxidation for production of retinoic acid: ethanol treatment in E7.5 mouse embryos reduced the amount of atRA produced, detected using transgenic reporter cell lines (Deltour et al, 1996).
Diethylaminobenzaldehyde (DEAB), an ALDH inhibitor: treatment of zebrafish embryos with DEAB reduced atRA signalling, detected using a transgenic reporter zebrafish model (Le et al, 2012).
Arnold SLM, Kent T, Hogarth CA, Griswold MD, Amory JK, Isoherranen N (2015) Pharmacological inhibition of ALDH1A in mice decreases all-trans retinoic acid concentrations in a tissue specific manner. Biochem Pharmacol 95: 177-192
Carlsen H, Ebihara K, Kuwata NH, Kuwata K, Aydemir G, Ruhl R, Blomhoff R (2021) A transgenic reporter mouse model for in vivo assessment of retinoic acid receptor transcriptional activation. Int J Vitam Nutr Res: 1-13
Chassot AA, Le Rolle M, Jolivet G, Stevant I, Guigonis JM, Da Silva F, Nef S, Pailhoux E, Schedl A, Ghyselinck NB, Chaboissier MC (2020) Retinoic acid synthesis by ALDH1A proteins is dispensable for meiosis initiation in the mouse fetal ovary. Sci Adv 6: eaaz1261
Morgenstern J, Fleming T, Kliemank E, Brune M, Nawroth P, Fischer A (2021) Quantification of All-Trans Retinoic Acid by Liquid Chromatography-Tandem Mass Spectrometry and Association with Lipid Profile in Patients with Type 2 Diabetes. Metabolites 11
Rossant J, Zirngibl R, Cado D, Shago M, Giguere V (1991) Expression of a retinoic acid response element-hsplacZ transgene defines specific domains of transcriptional activity during mouse embryogenesis. Genes Dev 5: 1333-1344