Ocean acidification affects the expression of neuroplasticity and neuromodulation markers in seabream.

A possible explanation for acidification-induced changes in fish behaviour is that acidification interferes with neurogenesis and modifies the plasticity of neuronal circuitry in the brain. We tested the effects on the olfactory system and brain of gilthead seabream (Sparus aurata) to 4 weeks' exposure to ocean acidification (OA). Olfactory epithelium (OE) morphology changed shortly after OA exposure and persisted over the 4 weeks. Expression of genes related to olfactory transduction, neuronal excitability, synaptic plasticity, GABAergic innervation, and cell proliferation were unchanged in the OE and olfactory bulb (OB) after 4 weeks' exposure. Short-term changes in the ionic content of plasma and extradural fluid (EDF) returned to control levels after 4 weeks' exposure, except for [Cl-], which remained elevated. This suggests that, in general, there is an early physiological response to OA and by 4 weeks a new homeostatic status is achieved. However, expression of genes involved in proliferation, differentiation and survival of undifferentiated neurons were modified in the brain. In the same brain areas, expression of thyroid hormone signalling genes was altered suggesting modifications in the thyroid-system may be linked to the changes in neuroplasticity and neurogenesis. Overall, the results of the current study are consistent with and effect of OA on neuroplasticity.

Non-mammalian models reveal the role of alternative ligands for thyroid hormone receptors.

Thyroid hormones, or THs, are well-known regulators of a wide range of biological processes that occur throughout the lifespan of all vertebrates. THs act through genomic mechanisms mediated by thyroid hormone receptors (TRs). The main product of the thyroid gland is thyroxine or T4, which can be further transformed by different biochemical pathways to produce at least 15 active or inactive molecules. T3, a product of T4 outer-ring deiodination, has been recognized as the main bioactive TH. However, growing evidence has shown that other TH derivatives are able to bind to, and/or activate TRs, to induce thyromimetic effects. The compiled data in this review points to at least two of these TR alternative ligands: TRIAC and T2. Taking this into account, non-mammalian models have proven to be advantageous to explore new TH derivatives with potential novel actions, prompting a re-evaluation of the role and mechanism of action of TR alternative ligands that were previously believed to be inactive. The functional implications of these ligands across different vertebrates may require us to reconsider current established notions of thyroid physiology.

Differential responses of the somatotropic and thyroid axes to environmental temperature changes in the green iguana.

Growth hormone (GH), together with thyroid hormones (TH), regulates growth and development, and has critical effects on vertebrate metabolism. In ectotherms, these physiological processes are strongly influenced by environmental temperature. In reptiles, however, little is known about the direct influences of this factor on the somatotropic and thyroid axes. Therefore, the aim of this study was to describe the effects of both acute (48h) and chronic (2weeks) exposure to sub-optimal temperatures (25 and 18°C) upon somatotropic and thyroid axis function of the green iguana, in comparison to the control temperature (30-35°C). We found a significant increase in GH release (2.0-fold at 25°C and 1.9-fold at 18°C) and GH mRNA expression (up to 3.7-fold), mainly under chronic exposure conditions. The serum concentration of insulin-like growth factor-I (IGF-I) was significantly greater after chronic exposure (18.5±2.3 at 25°C; 15.92±3.4 at 18°C; vs. 9.3±1.21ng/ml at 35°C), while hepatic IGF-I mRNA expression increased up to 6.8-fold. Somatotropic axis may be regulated, under acute conditions, by thyrotropin-releasing hormone (TRH) that significantly increased its hypothalamic concentration (1.45 times) and mRNA expression (0.9-fold above control), respectively; and somatostatin (mRNA expression increased 1.0-1.2 times above control); and under chronic treatment, by pituitary adenylate cyclase-activating peptide (PACAP mRNA expression was increased from 0.4 to 0.6 times). Also, it was shown that, under control conditions, injection of TRH stimulated a significant increase in circulating GH. On the other hand, while there was a significant rise in the hypothalamic content of TRH and its mRNA expression, this hormone did not appear to influence the thyroid axis activity, which showed a severe diminution in all conditions of cold exposure, as indicated by the decreases in thyrotropin (TSH) mRNA expression (up to one-eight of the control), serum T4 (from 11.6±1.09 to 5.3±0.58ng/ml, after 2weeks at 18°C) and T3 (from 0.87±0.09 to 0.05±0.01ng/ml, under chronic conditions at 25°C), and Type-2 deiodinase (D2) activity (from 992.5±224 to 213.6±26.4fmolI(125)T4/mgh). The reduction in thyroid activity correlates with the down-regulation of metabolism as suggested by the decrease in the serum glucose and free fatty acid levels. These changes apparently were independent of a possible stress response, at least under acute exposure to both temperatures and in chronic treatment to 25°C, since serum corticosterone had no significant changes in these conditions, while at chronic 18°C exposure, a slight increase (0.38 times above control) was found. Thus, these data suggest that the reptilian somatotropic and thyroid axes have differential responses to cold exposure, and that GH and TRH may play important roles associated to adaptation mechanisms that support temperature acclimation in the green iguana.

3,5-Diiodothyronine-mediated transrepression of the thyroid hormone receptor beta gene in tilapia. Insights on cross-talk between the thyroid hormone and cortisol signaling pathways.

T3 and cortisol activate or repress gene expression in virtually every vertebrate cell mainly by interacting with their nuclear hormone receptors. In contrast to the mechanisms for hormone gene activation, the mechanisms involved in gene repression remain elusive. In teleosts, the thyroid hormone receptor beta gene or thrb produces two isoforms of TRß1 that differ by nine amino acids in the ligand-binding domain of the long-TRß1, whereas the short-TRß1 lacks the insert. Previous reports have shown that the genomic effects exerted by 3,5-T2, a product of T3 outer-ring deiodination, are mediated by the long-TRß1. Furthermore, 3,5-T2 and T3 down-regulate the expression of long-TRß1 and short-TRß1, respectively. In contrast, cortisol has been shown to up-regulate the expression of thrb. To understand the molecular mechanisms for thrb modulation by thyroid hormones and cortisol, we used an in silico approach to identify thyroid- and cortisol-response elements within the proximal promoter of thrb from tilapia. We then characterized the identified response elements by EMSA and correlated our observations with the effects of THs and cortisol upon expression of thrb in tilapia. Our data show that 3,5-T2 represses thrb expression and impairs its up-regulation by cortisol possibly through a transrepression mechanism. We propose that for thrb down-regulation, ligands other than T3 are required to orchestrate the pleiotropic effects of thyroid hormones in vertebrates.

The variable region of iodothyronine deiodinases directs their catalytic properties and subcellular localization.

The stereospecific removal of iodine from thyroid hormones is an essential first step for T3 action and is catalyzed by three different deiodinases: D2 and D3 remove iodine only from the outer or inner ring, respectively, whereas D1 catalyzes both pathways. We used in silico predictions from vertebrate deiodinase sequences to identify two domains: the N-terminal variable region (VR) containing the transmembrane, hinge and linker domains, and the conserved or globular region (CR). Given the high sequence and structural identity of the CR among paralogs as well as of the VR among orthologs but not paralogs, we hypothesized that both the catalytic properties and the subcellular localization rely on the VR. We used shark D2 and D3 as templates to build the chimeric enzymes D2VR/D3CR and D3VR/D2CR. Biochemical characterization revealed that D3VR/D2CR has inner-ring deiodination activity and T3 as preferred substrate, whereas D2VR/D3CR showed no deiodinating activity. Also, D2VR/D3CR and D3VR/D2CR reside in the endoplasmic reticulum and plasmatic membrane, respectively, as do their D2 and D3 wild-type counterparts. We conclude that the VR determines the subcellular localization and is critical in defining the catalytic properties and activity of thyroid hormone deiodinases.

3,5-Diiodothyronine (T2) is on a role. A new hormone in search of recognition.

Thyroid hormone (TH) actions are mediated by triiodothyronine (T3), which acts by binding to the TH receptors (TRs). Since TH exert pleiotropic effects, interest has grown in identifying other possible bioactive thyronines that could explain their diversity of functions. Accordingly, 3,5-diiodothyronine (T2) has been shown to be bioactive. In mammals, T2 regulates mRNA expression of several T3-regulated genes, but doses up to 100-fold greater than those of T3 were required to generate comparable effects. In teleosts, T2 and T3 regulate gene expression in vivo with equivalent potency. Furthermore, in vivo and in vitro studies support the notion that T2 binds to and activates a specific, long TRß1 isoform that contains a nine amino acid insert at the beginning of the ligand binding domain, whereas T3 can interact also with a different TRß1 isoform that lacks this insert. Similarly, T2 and T3 differentially regulate long- and short-TRß1 expression, respectively, strongly suggesting a different signaling pathway for each hormone, at least in the species that express both receptors. In vivo, T2 effectively triggers a burst of body growth in tilapia by interacting with the long TRß1 isoform, supporting the notion that T2 is physiologically relevant in this species. Current knowledge of T2 effects and action mechanisms lead us to propose that there is an extra level in the thyroid hormone signaling cascade, and that T2 is produced and regulated specifically for this purpose.

Iodothyronine deiodinases: a functional and evolutionary perspective.

From an evolutionary perspective, deiodinases may be considered pivotal players in the emergence and functional diversification of both thyroidal systems (TS) and their iodinated messengers. To better understand the evolutionary pathway and the concomitant functional diversification of vertebrate deiodinases, in the present review we summarized the highlights of the available information regarding this ubiquitous enzymatic component that represents the final, common physiological link of TS. The information reviewed here suggests that deiodination of tyrosine metabolites is an ancient feature of all chordates studied to date and consequently, that it precedes the integration of the TS that characterize vertebrates. Phylogenetic analysis presented here points to D1 as the oldest vertebrate deiodinase and to D2 as the most recent deiodinase gene, a hypothesis that agrees with the notion that D2 is the most specialized and finely regulated member of the family and plays a key role in vertebrate neurogenesis. Thus, deiodinases seem to be major participants in the evolution and functional expansion of the complex regulatory network of TS found in vertebrates.