Regulation of the corticosteroid signalling system in rainbow trout HPI axis during confinement stress (original) (raw)
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General and Comparative Endocrinology, 2012
Cortisol, the primary circulating corticosteroid in teleosts, is elevated during stress following activation of the hypothalamus-pituitary-interrenal (HPI) axis. Cortisol exerts genomic effects on target tissues in part by activating glucocorticoid receptors (GR). Despite a well-established negative feedback loop involved in plasma cortisol regulation, the role of GR in the functioning of the HPI axis during stress in fish is still unclear. We used mifepristone (a GR antagonist) to suppress GR signaling in rainbow trout (Oncorhynchus mykiss) and assessed the resultant changes to HPI axis activity. We show for the first time that mifepristone caused a functional knockdown of GR by depleting protein expression 40-75%. The lower GR protein expression corresponded with a compensatory up-regulation of GR mRNA levels across tissues. Mifepristone treatment completely abolished the stressor-induced elevation in plasma cortisol and glucose levels seen in the control fish. A reduction in corticotropin-releasing factor (CRF) mRNA abundance in the hypothalamic preoptic area was also observed, suggesting that GR signaling is involved in maintaining basal CRF levels. We further characterized the effect of mifepristone treatment on the steroidogenic capacity of interrenal tissue in vitro. A marked reduction in cortisol production following adrenocorticotropic hormone stimulation of head kidney pieces was observed from mifepristone treated fish. This coincided with the suppression of steroidogenic acute regulatory protein, but not P450 side chain cleavage mRNA abundances. Overall, our results underscore a critical role for central and peripheral GR signaling in the regulation of plasma cortisol levels during stress in fish.
Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2013
The present study describes the transcriptional levels of the corticosteroid receptors (CRs) GR1, GR2 and MR in the different organs of the rainbow trout (Oncorhynchus mykiss) in response to a slow release of cortisol, throughout a 10-day period. We show that after short term (1 day after cortisol implantation), when the plasma levels of cortisol emulate an acute stress, the GR2 and MR expression levels were upregulated in the brain and head kidney tissues. This result reflects the role of these organs as regulators of the stress response. In general, the rest of the organs, especially gills, intestine, liver, muscle and spleen, showed decreased transcriptional levels of GR1, GR2 and MR, along with the highest plasma cortisol levels. At day 5 after cortisol implantation, when cortisol levels emulate a chronic stress, the most affected organs were gills and skin, where an upregulation of the CRs was found. In the recovery period, when cortisol levels were basal (day 10), we still found changes in the transcriptional levels of the CRs in gills, spleen and gonads. The cortisol increase at days 1 and 5 after implantation is accompanied by high plasma glucose concentrations, supporting the role of cortisol on carbohydrate metabolism. However, after 10 days of implantation, glucose returned to control levels suggesting a trade-off on the steady state of the metabolic function. We also observed increased hematocrit and hemoglobin at day 1, indicating a cortisol-induced higher metabolic demand involving an increase in oxygen transport efficiency. Our results demonstrate that increased plasma cortisol induced by a slow-release implant of cortisol mimics the overall effects of stress and affects the expression of the three CRs, generating different transcriptional patterns in a time-and organ-specific manner.
Journal of cellular biochemistry, 2016
Cortisol is an essential regulator of neuroendocrine stress responses in teleosts. Cortisol predominantly affects target tissues through the genomic pathway, which involves interacting with cytoplasmic glucocorticoid receptors and, thereby, modulating stress-response gene expressions. Cortisol also produces rapid effects via non-genomic pathways, which do not involve gene transcription. Although cortisol-mediated genomic pathways are well documented in teleosts, non-genomic pathways are not fully understood. Moreover, no studies have focused on the contribution of non-genomic cortisol pathways in compensatory stress responses in fish. In this study, rainbow trout (Oncorhynchus mykiss) skeletal myotubes were stimulated with physiological concentrations of cortisol and cortisol-BSA, a membrane-impermeable agent, resulting in an early induction of reactive oxygen species (ROS). This production was not suppressed by transcription or translation inhibitors, suggesting non-genomic pathway...
Corticosteroid receptors involved in stress regulation in common carp, Cyprinus carpio
Journal of Endocrinology, 2008
In higher vertebrates, mineralo- (aldosterone) and glucocorticoids (cortisol/corticosterone) exert their multiple actions via specific transcription factors, glucocorticoid (GR) and mineralocorticoid (MR) receptors. Teleostean fishes lack aldosterone and mineral regulatory processes seem under dominant control by cortisol. Despite the absence of the classical mineralocorticoid aldosterone, teleostean fishes do have an MR with cortisol and possibly 11-deoxycorticosterone (DOC) (as alternative for aldosterone) as predominant ligands. We studied corticoid receptors in common carp (Cyprinus carpio L). Through homology cloning and bioinformatic analysis, we found duplicated GR genes and a single MR gene. The GR genes likely result from a major genomic duplication event in the teleostean lineage; we propose that the gene for a second MR was lost. Transactivation studies show that the carp GRs and MR have comparable affinity for cortisol; the MR has significantly higher sensitivity to DOC,...
Fish Physiology and Biochemistry, 1994
Plasma cortisol levels and the number (Nmax) and affinity (Kd) of specific hepatic cortisol-binding sites were determined in rainbow trout subjected to chronic confinement stress for 14 days. Confinement significantly elevated plasma cortisol levels to 47.3 ± 13.5 ng ml-I within 24h and although levels declined to 8.0 3.0 ng ml-1 after 14 days, they were significantly higher throughout than levels in unstressed control fish (< 2.0 ng ml-1). There was a 60Oo reduction in cytosolic Nmax in stressed fish during the first 24h of confinement (35.8 ± 7.9 cf. 129.0 15.2 fmol mg -l protein), a decline which was sustained at 7 days after the onset of stress but, although numbers of binding sites in the liver of stressed fish were still lower than in unstressed fish, the difference was no longer significant after 14 days of confinement. There was an accompanying significant rise in the Kd of cortisol binding in stressed fish during confinement, from 4.0 ± 0.6 nM at time 0 to 8.4 ± 0.8 nM after 24 h confinement. This increment in Kd was sustained at a level significantly higher than in control fish throughout the 14 day confinement period, despite marked reductions in cortisol levels and increases in Nmax in stressed fish. Throughout the study, specific binding of cortisol could not be consistently detected in high-salt nuclear extracts from stressed or unstressed fish, suggesting either that highaffinity binding sites for cortisol were absent from these preparations, that receptors were present but unable to interact with ligand because they were occupied, or that receptors were present but not being extracted. These possibilities were investigated using a range of extraction procedures, by varying the temperature of incubation, by employing dexamethasone as ligand and by examining binding in purified, intact, nuclei. Estradiol was employed as a methodological control throughout and substantial amounts of specific estradiol binding were detected in all compartments and preparations. Specific cortisol-binding sites were detected in intact nuclei of both stressed and unstressed fish, at levels an order of magnitude lower than estradiol binding in the same preparations. These data demonstrate that activation of the pituitary-interrenal axis leads to significant changes in the nature of target-tissue binding of cortisol in rainbow trout, and reveal a clear difference in the subcellular distribution of binding-sites for estradiol and cortisol, which reflects the situation in mammalian tissues.
The glucocorticoid stress response is repeatable between years in a wild teleost fish
Journal of Comparative Physiology A, 2011
Patterns of glucocorticoid (GC) hormone regulation exhibit considerable inter-individual variation that is often examined relative to individual traits and fitness measures. Although stress-induced GC concentrations are repeatable within an individual in captive populations, this assumption remains untested in wild animals in their natural environment across longer time periods. We assessed the repeatability of baseline and post-stress GC concentrations in a wild teleost fish. Largemouth bass (Micropterus salmoides) were captured and subjected to a standard stress protocol and then stocked into a small research lake. Upon recapture by angling up to 1 year later (n = 26), fish were re-sampled following identical methods. After controlling for a strong effect of water temperature, we confirmed repeatability of post-stress cortisol concentrations despite stress presumed to accompany relocation. We documented no consistency in baseline GC concentrations. This study serves as an important validation for the use of post-stress cortisol concentrations as an individual trait. However, the effect size of repeatability was lower than that found in other taxa. Results also bring forth the reality that environmental variables such as temperature must be considered in studies where these factors can vary, such as when sampling wild animals at liberty.
Cortisol in Correlation to Other Indicators of Fish Welfare
Chapter in Corticosteroids, IntechOpen , London, UK, 2018
Cortisol is the major corticosteroid in teleost fish, secreted and released by interrenal cells of the head kidney during activation of the hypothalamic-pituitary-interrenal (HPI) axis. Although cortisol is universally recognized as a key mediator of stress-associated responses, other hormones are also involved in the stress response, e.g., arginine vasoto-cin (AVT), isotocin (IT), urotensins, dopamine, serotonin or β-endorphin. Cortisol affects AVT and IT secretion from nerve endings in gilthead sea bream (Sparus aurata) and round goby (Neogobius melanostomus). Moreover, it is pointed out that different mechanisms are involved in the regulation of AVT and IT release from the hypothalamic-pituitary complex in round goby. In the case of AVT, both genomic and nongenomic pathways are mediating the effect of cortisol while in the case of IT, it is only the nongenomic pathway. In turn, urotensin I instead of corticotropin-releasing factor (CRF) may contribute to the regulation of HPI axis and regulate AVT in Sparus aurata. In this species, urotensin II together with AVT and IT may control stress response to different salinities. Therefore, AVT, IT and urotensins, and their interactions with cortisol, seem to be significant in response to stress in fish.
Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2003
The purpose of this study was to characterize the association between hepatic heat shock protein 70 (hsp70) and the glucocorticoid receptor in rainbow trout that were exposed to heat stress, cortisol, and b-naphthoflavone. This study is the first to document that the glucocorticoid receptor complex in rainbow trout hepatic tissues contains hsp70. Heat stress significantly increased levels of total cellular hsp70, and by discerning the association of hsp70 with the glucocorticoid receptor, we demonstrated that heat stress significantly increased the amount of hsp70 not bound to the glucocorticoid receptor, while significantly decreasing the amount of hsp70 bound to the glucocorticoid receptor. By calculating the ratio of hsp70 bound to the glucocorticoid receptor, to the total number of glucocorticoid receptors, stress (heat stress and cortisol-treatment) promoted the association of hsp70 with the glucocorticoid receptor. These findings demonstrate a functional and structural link between hsp70 and the glucocorticoid receptor in rainbow trout, and raise questions regarding the existence of a complex, interrelated stress response that spans all levels of biological organization within the whole animal.
General and Comparative Endocrinology, 2014
Based on previous studies we hypothesize that under stress conditions catecholamine-induced hyperglycemia contributes to enhance cortisol production in head kidney of rainbow trout. Therefore, treatment with propranolol (b-adrenoceptor blocker) should reduce the hyperglycemia elicited by stress and, therefore, we expected reduced glucosensing response and cortisol production in head kidney. Propranolol treatment was effective in blocking most of the effects of catecholamines in liver energy metabolism resulting in a lower glycemia in stressed fish. The decreased glycemia of stressed fish treated with propranolol was observed along with reduced transcription of genes involved in the cortisol synthetic pathway, which supports our hypothesis. However, changes in putative glucosensing parameters assessed in head kidney were scarce and in general did not follow changes noted in glucose levels in plasma. Furthermore, circulating cortisol levels did not change in parallel with changes in glycemia. As a whole, the present results suggest that glycemia could participate in the regulation of cortisol synthetic pathways but other factors are also likely involved. Propranolol effects on trout stress response were different depending on time passed after stress onset; the direct or indirect involvement of catecholaminergic response in the regulation of cortisol production and release deserves further investigation.
Condition dependent intra-individual repeatability of stress-induced cortisol in a freshwater fish
Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2012
The glucocorticoid (GC) stress response is thought to be an individual trait associated with behaviour and life history strategies. Studies exploring such relationships typically assume measured hormone values to be repeatable within an individual. However, repeatability of GCs has proven variable in wild animals and underlying reasons remain unknown. We assessed individual repeatability of circulating stress-induced cortisol, the primary GC in teleost fish, and glucose concentrations in a wild teleost fish held under consistent laboratory conditions. We also tested the hypothesis that the magnitude of intra-individual variability in stressinduced cortisol concentrations ("cortisol variability") is influenced by body condition. Wild-caught bluegill sunfish (Lepomis macrochirus) were subjected to repeated standardized stressors and blood sampled (3 times over 6 days) once cortisol concentrations peaked. Various indicators of fish condition, both whole body and physiological, were also measured. Overall, stress-induced circulating cortisol concentrations were repeatable but stress-induced glucose was not. Cortisol variability was related to Fulton's condition factor and size (eviscerated mass) where smaller fish in poor condition exhibited increased cortisol variability. The findings have implications for the interpretation of studies that examine correlates of GC concentrations as they suggest consistency in stress responsiveness is influenced by factors such as size and condition.