Upstream eventInhibition, Deiodinase 2
Decreased, Triiodothyronine (T3) in serum
Key Event Relationship Overview
AOPs Referencing Relationship
|AOP Name||Adjacency||Weight of Evidence||Quantitative Understanding|
|Deiodinase 2 inhibition leading to reduced young of year survival via posterior swim bladder inflation||adjacent|
|Deiodinase 2 inhibition leading to reduced young of year survival via anterior swim bladder inflation||adjacent|
|fathead minnow||Pimephales promelas||NCBI|
Life Stage Applicability
Key Event Relationship Description
Iodothyronine deiodinase or DIO is a peroxidase enzyme that is involved in the activation or deactivition of thyroid hormones. Currently, three types of iodothyronine deiodinases (DIO1-3) have been described in vertebrates that locally activate or inactivate THs and are therefore important mediators of TH action. All deiodinases are integral membrane proteins of the thioredoxin superfamily that contain the amino acid selenocysteine in their catalytic centre. DIO1 and DIO2 are capable of converting T4 into the more biologically active T3. DIO3 on the other hand converts T4 and T3 to the inactive forms of THs. The inhibition of DIO 1 and 2 enzymes results in decreased serum T3 levels and decreased T3 levels at the site of action.
Evidence Supporting this KER
Inhibition of DIO2 activity is widely accepted to directly decrease T3 levels, since the conversion of T4 to T3 is inhibited. The importance of DIO2 inhibition in altering serum T3 levels depends on the relative role of different deiodinases in regulating serum versus tissue T3 levels and in negative feedback within the HPT axis. Both aspects appear to vary among vertebrate taxa.
Inhibition of DIO2 activity is widely accepted to directly decrease T3 levels, since the conversion of T4 to T3 is inhibited.
- Houbrechts et al. (2016) developed a Dio2 knockout and confirmed both the absence of the full length Dio2 protein in the liver and the dramatical decrease of T4 activating enzyme activity in liver, brain and eyes. Finally, they found decreased levels of T3 in liver, brain and eyes.
- Winata et al. (2009, 2010) reported reduced pigmentation, otic vesicle length and head-trunk angle in DIO1+2 and DIO2 knockdown fish. These effects were rescued after T3 supplementation but not by T4 supplementation, confirming that decreased T3 levels were at the basis of the observed effects.
- In the study of Cavallin et al. (2017) fathead minnow larvae were exposed to IOP, a model iodothyronine deiodinase inhibitor that is assumed to inhibit all three deiodinase enzymes (DIO1,2,3). Transcriptional analysis showed that especially DIO2, but also DIO3 mRNA levels (in some treatments), were increased in 10 to 21 day old larvae exposed to IOP as of the age of 6 days. This suggests that IOP effectively inhibited DIO2 and DIO3 in the larvae and that mRNA levels increased as a compensatory response. The authors also observed pronounced decreases of whole body T3 concentrations and increases of whole body T4 concentrations.
- Stinckens et al. (submitted) showed that IOP reduced T3 levels in zebrafish in 21 and 32 day old larvae that had been exposed starting from fertilization.
Uncertainties and Inconsistencies
In DIO2 knockout mice it seemed that the negative feedback system was blocked resulting in increased levels of T4 and TSH and in normal rather than decreased T3 levels compared to WT.
In the study of Cavallin et al. (2017) fathead minnow embryos were exposed to IOP, a model iodothyronine deiodinase inhibitor that is assumed to inhibit all three deiodinase enzymes (DIO1,2,3). The authors observed increased whole body T3 concentrations in 4 and 6 day old embryos, while they observed decreased T3 concentrations in 10 to 21 day old larvae exposed to IOP as of the age of 6 days. One possible explanation for the elevated T3 concentrations may be the potential impact of IOP exposure on DIO3. DIO3 is an inactivating enzyme that removes iodine from the inner ring of both T4 and T3, resulting in reverse T3 (rT3) and 3,5-diiodo-L-thyronine (T2), respectively (Bianco and Kim, 2006). Maternal sources of thyroid hormones are known to include both T4 and T3 (Power et al., 2001; Walpita et al., 2007). Consequently, reduced conversion of maternal T3 to inactive forms may be one plausible explanation for the increase. Another explanation may result from the role of deiodinases in the negative feedback system of the HPT axis. Inibition of deiodinase (unclear which isoforms) may block the negative feedback system and result in increased release of T4. Increased levels of T4 were indeed observed by Cavallin et al. (2017).
Quantitative Understanding of the Linkage
Known modulating factors
Known Feedforward/Feedback loops influencing this KER
Thyroid hormone levels are regulated via negative feedback, influencing this KER. Additionally, deiodinases regulate the activity of thryoid hormones, not only in serum and target organs, but also in the thryoid gland. Deiodinases themselves are known to be involved in the negative feedback system that results in increased TSH levels when the levels of T4 (and also T3) in serum are low (Schneider et al., 2001), resulting in an even more complicated impact on this KER. Increased TSH levels then stimulate increased T4 release from the thyroid gland, resulting in a compensatory increase of serum T4 levels. In DIO2 knockout mice it seemed that the negative feedback system was blocked resulting in increased levels of T4 and TSH and in normal rather than decreased T3 levels compared to WT. By inhibiting DIO1 using a PTU exposure, Schneider et al. (2001) showed that DIO2 played a role in the increased TSH levels in response to T3 or T4 injection.
Domain of Applicability
Mol et al. (1998) concluded that deiodinases in teleosts were more similar to mammalian deiodinases than had been generally accepted, based on the similarities in susceptibility to inhibition and the agreement of the Km values.
There appear to be differences among vertebrate classes relative to the role of the different deiodinase isoforms in regulating thyroid hormone levels. Maia et al. (2005) determined that in a normal physiological situation in humans the contribution of DIO2 to plasma T3 levels is twice that of DIO1. A DIO2 knockout (KO) mouse however showed a very mild gross phenotype with only mild growth retardation in males (Schneider et al., 2001). It seemed that by blocking the negative feedback system, DIO2 KO resulted in increased levels of T4 and TSH and in normal rather than decreased T3 levels compared to WT. Potential differences in the role of the deiodinase isoforms in the negative feedback system and the final consequences for TH levels across vertebrates is currently not entirely clear. These differences make it difficult to exactly evaluate the importance of DIO2 in regulating serum/tissue T3 levels across vertebrates.
Bianco, A.C., Kim, B.W., 2006. Deiodinases: implications of the local control of thyroid hormone action. Journal of Clinical Investigation 116, 2571-2579.
Cavallin, J.E., Ankley, G.T., Blackwell, B.R., Blanksma, C.A., Fay, K.A., Jensen, K.M., Kahl, M.D., Knapen, D., Kosian, P.A., Poole, S.T., Randolph, E.C., Schroeder, A.L., Vergauwen, L., Villeneuve, D.L., 2017. Impaired swim bladder inflation in early life stage fathead minnows exposed to a deiodinase inhibitor, iopanoic acid. Environmental Toxicology and Chemistry 36, 2942-2952.
Houbrechts, A.M., Delarue, J., Gabriels, I.J., Sourbron, J., Darras, V.M., 2016. Permanent Deiodinase Type 2 Deficiency Strongly Perturbs Zebrafish Development, Growth, and Fertility. Endocrinology 157, 3668-3681.
Maia, A.L., Kim, B.W., Huang, S.A., Harney, J.W., Larsen, P.R., 2005. Type 2 iodothyronine deiodinase is the major source of plasma T-3 in euthyroid humans. Journal of Clinical Investigation 115, 2524-2533.
Mol, K.A., Van der Geyten, S., Burel, C., Kuhn, E.R., Boujard, T., Darras, V.M., 1998. Comparative study of iodothyronine outer ring and inner ring deiodinase activities in five teleostean fishes. Fish Physiology and Biochemistry 18, 253-266.
Power, D.M., Llewellyn, L., Faustino, M., Nowell, M.A., Bjornsson, B.T., Einarsdottir, I.E., Canario, A.V., Sweeney, G.E., 2001. Thyroid hormones in growth and development of fish. Comp Biochem Physiol C Toxicol Pharmacol 130, 447-459.
Schneider, M.J., Fiering, S.N., Pallud, S.E., Parlow, A.F., St Germain, D.L., Galton, V.A., 2001. Targeted disruption of the type 2 selenodeiodinase gene (D102) results in a phenotype of pituitary resistance to T-4. Molecular Endocrinology 15, 2137-2148.
Stinckens, E., Vergauwen, L., Blackwell, B.R., Ankley, G.T., Villeneuve, D.L., Knapen, D., The effect of thyroperoxidase and deiodinase inhibition on anterior swim bladder inflation in the zebrafish. Environmental Science & Technology submitted.
Walpita, C.N., Van der Geyten, S., Rurangwa, E., Darras, V.M., 2007. The effect of 3,5,3'-triiodothyronine supplementation on zebrafish (Danio rerio) embryonic development and expression of iodothyronine deiodinases and thyroid hormone receptors. Gen Comp Endocrinol 152, 206-214.
Winata, C.L., Korzh, S., Kondrychyn, I., Korzh, V., Gong, Z. 2010. The role of vasulature and blood circulation in zebrafish swim bladder development. Dev. Biol. 10:3.
Winata, C.L., Korzh, S., Kondrychyn, I., Zheng, W., Korzh, V., Gong, Z. 2009. Development of zebrafish swimbladder: the requirement of Hedgehog signaling in specification and organization of the three tissue layers. Dev. Biol.331, 222–236, http://dx.doi.org/10.1016/j.ydbio.2009.04.035.