Increased Corticosterone Levels Research Articles

Activation of Local 11β-Hydroxysteroid Dehydrogenase Type 1 by Diosmetin Enhances Endogenous Glucocorticoid Levels to Alleviate Skin Inflammation: Insights Into a Novel Therapeutic Strategy for Atopic Dermatitis.

Glucocorticoids (GCs) are synthesised de novo by peripheral tissues and the adrenal cortex of the hypothalamic-pituitary-adrenal axis. Skin expresses an enzyme called 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), which reduces cortisone to the active hormone cortisol which activates GC receptors. 11β-HSD1 plays a significant role in alleviating atopic inflammation through the elevation of the concentrations of active GC in the skin. This study aimed to investigate the role of diosmetin as an activator of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1). In human keratinocytes, diosmetin was found to upregulate 11β-HSD1 protein expression and cortisol levels, as well as the transcriptional expression of 11β-HSD1 mRNA. However, this upregulation of 11β-HSD1 mRNA was abrogated in keratinocytes transfected with 11β-HSD1 small interfering RNA (siRNA). In an atopic dermatitis (AD) murine model, topical administration of diosmetin significantly attenuated basal transepidermal water loss (TEWL) and the Eczema Area and Severity Index (EASI), while enhancing stratum corneum (SC) hydration. Diosmetin also increased corticosterone levels in the SC and upregulated 11β-HSD1 expression in both the serum and epidermis. Furthermore, diosmetin treatment led to a marked reduction in serum immunoglobulin E (IgE) and tumour necrosis factor-α (TNF-α) levels, and suppressed mRNA expression of thymic stromal lymphopoietin (TSLP), interleukin-1β (IL-1β), IL-4, and IL-13 in the epidermis. Collectively, these findings suggest that diosmetin promotes the endogenous activation of glucocorticoids via local 11β-HSD1 activation, underscoring its potential as a novel topical therapeutic agent for the management of inflammatory skin disorders, such as AD.

Acute and chronic cannabis vapor exposure influences basal and stress-induced release of glucocorticoids in male and female rats.

Management of stress and anxiety is often listed as the primary motivation behind cannabis use. Human research has found that chronic cannabis use is associated with increased basal cortisol levels but blunted neuroendocrine responses to stress. Preclinical research has demonstrated mixed effects of Δ9-tetrahydrocannabinol (THC; the psychoactive constituent of cannabis), much of which is suggestive of dose-dependent effects; however, the predominance of this work has employed an injection method to deliver cannabis. As inhalation is the most common route of administration in humans, we employed a translationally relevant model of inhaled cannabis vapor exposure to help characterize the extent to which acute and chronic cannabis exposure modulates neuroendocrine responses to stress. Male and female Sprague-Dawley rats were acutely (single day) or chronically (10 days) exposed to cannabis or vehicle vapor, and the stress hormone, corticosterone, was analyzed prior to and following an acute 30-min restraint stress. Our results indicate that initial exposure to the vapor chambers, regardless of vehicle or cannabis exposure, is sufficient to elevate corticosterone levels in male and female rodents. Further, acute cannabis exposure was capable of increasing corticosterone levels in both male and female rats, however, this effect was modified by the habituation to the vapor chambers differentially in males and females. Regardless of sex, chronic cannabis exposure is sufficient to both elevate basal corticosterone levels and blunt stress-induced increases in corticosterone following a restraint stressor. Collectively, these data help characterize the impacts of cannabis vapor exposure on basal and stress-induced activation of the hypothalamic-pituitary-adrenal axis.

Dim blue light at night worsens high-fat diet-induced kidney damage via increasing corticosterone levels and modulating the expression of circadian clock genes.

Obesity is a contributing factor that increases the likelihood of developing chronic kidney disease. In recent years, studies have found that light pollution worldwide promoted obesity, which was known to be a consequence of circadian rhythm disruption. Nevertheless, the impact of light pollution on kidney disease associated with obesity remains mostly unknown, and potential processes have been minimally investigated. Herein, we fed mice a high-fat diet and gave them dim white (dWL), blue (dBL), green (dGL), and red (dRL) light for 12 weeks. Our results showed that both dWL and dBL tended to be more susceptible to damage to kidney dysfunction caused by a high-fat diet compared to LD, with more pronounced changes in dBL. The analysis of kidney found that dBL activated the TGF-β1/Smad signaling pathway to promote epithelial-mesenchymal transition (EMT) in the kidney. Additionally, dBL activated the NF-κB signaling pathway and resulted in elevated protein levels of TLR4, p-IκB, p-P65, and TNF-α. Furthermore, dBL increased BAX protein levels and decreased BCL2 protein levels. At the same time, dBL affected the Keap1/Nrf2/HO-1 signaling pathway, elevating KEAP1 and decreasing NRF2 and HO-1 protein levels. We were surprised to find that dBL altered the expression of the circadian clock genes, resulting in a decrease in the positively regulated genes Bmal1, Clock, and an increase in the negatively regulated genes Per1, Per2, Per3. Mechanistically, dBL increased plasma CORT levels as well as decreased renal GR α expression, and in vitro experiments showed that the circadian clock genes were altered by the addition of CORT and returned to normal levels after the addition of the GR inhibitor RU486. Consequently, dBL can exacerbate renal injury by elevating plasma CORT levels and altering rhythmic changes by acting on the biological clock via GR.

Maternal sucrose consumption alters steroid levels in the mother, placenta, and fetus.

Maternal diet has long-term effects on offspring brain development and behavior. Sucrose (table sugar) intakes are high in modern diets, but it is not clear how a maternal high-sucrose diet (HSD) affects the offspring. In rats, a maternal HSD (26% of calories from sucrose, which is human-relevant) alters maternal metabolism and brain and also alters adult offspring endocrinology and behavior in a sex-specific manner. Maternal sucrose intake increases corticosterone levels in adult female offspring and increases motivation for a sugar reward in adult male offspring. Here, to identify possible underlying mechanisms, we examined how a maternal HSD affects steroids in the dam, placenta, and fetus at embryonic day 19.5 using liquid chromatography tandem mass spectrometry. Maternal sucrose intake increased glucocorticoids (11-deoxycorticosterone and 11-dehydrocorticosterone) and tended to increase the mineralocorticoid aldosterone in maternal serum. In the placenta, maternal sucrose intake decreased androstenedione and testosterone. Maternal HSD increased aldosterone in the fetal blood. Similarly, in the fetal brain, maternal high-sucrose intake increased aldosterone in the medial prefrontal cortex and nucleus accumbens, decreased testosterone in the nucleus accumbens, and decreased corticosterone the orbital cortex. In addition, the 11-dehydrocorticosterone/corticosterone and aldosterone/corticosterone ratios were increased in most examined brain regions. Lastly, maternal HSD increased 11-dehydrocorticosterone and aldosterone in the amniotic fluid. In summary, we found dramatic and widespread changes in maternal, placental, and fetal steroids that might mediate the long-term effects of maternal sucrose consumption on adult offspring neuroendocrinology and behavior.

Concurrent stress modulates the acute and post-acute effects of psilocybin in a sex-dependent manner.

There is renewed interest in psychedelics, such as psilocybin, as therapies for multiple difficult-to-treat psychiatric disorders. Even though psychedelics can induce highly pleasant or aversive experiences, depending on multiple personal and environmental factors, there is little research into how such experiences impact post-acute mood-altering actions. Here we aimed at offsetting this gap. First, we tested whether acute psilocybin effects differed between sexes. Adult male and female C57BL/6J mice received saline or psilocybin (5mg/kg;i.p.), and head-twitch response (HTR) frequency was quantified. Notably, while psilocybin increased HTR frequency in both sexes, the effect was greater in females. We then tested if stress exposure during acute drug effects impacted post-acute psilocybin actions. Following drug treatment, mice were returned to their homecage or restrained for 1h. Anxiety- and depressive-like behaviors were assessed starting 24h following drug administration, using the marble burying, novelty-suppressed feeding, and splash tests. Psilocybin induced anxiolytic-, but not antidepressant-like, which were fully blocked by stress in males, but only partially so in females. Lastly, we assessed the acute stress-psilocybin interaction on plasma corticosterone levels in a separate cohort of mice, treated as above. Both stress and psilocybin independently increased corticosterone levels, without additive or interactive effects being observed for either sex. Our data reveals the role of sex and peri-acute negative experiences in the acute and post-acute actions of psilocybin. These findings underline the importance of non-pharmacological factors, such as the quality of the psychedelic experience, in the mood-altering effects of psychedelics, holding significant for both their therapeutic and recreational use.

Mechanisms of Heat Stress on Neuroendocrine and Organ Damage and Nutritional Measures of Prevention and Treatment in Poultry.

Heat stress (HS) due to high temperatures has adverse effects on poultry, including decreased feed intake, lower feed efficiency, decreased body weight, and higher mortality. There are complex mechanisms behind heat stress in poultry involving the neuroendocrine system, organ damage, and other physiological systems. HS activates endocrine glands, such as the pituitary, adrenal, thyroid, and gonadal, by the action of the hypothalamus and sympathetic nerves, ultimately causing changes in hormone levels: HS leads to increased corticosterone levels, changes in triiodothyronine and thyroxine levels, decreased gonadotropin levels, reduced ovarian function, and the promotion of catecholamine release, which ultimately affects the normal productive performance of poultry. Meanwhile, heat stress also causes damage to the liver, lungs, intestines, and various immune organs, severely impairing organ function in poultry. Nutrient additives to feed are important measures of prevention and treatment, including natural plants and extracts, probiotics, amino acids, and other nutrients, which are effective in alleviating heat stress in poultry. Future studies need to explore the specific mechanisms through which heat stress impacts the neuroendocrine system in poultry and the interrelationships between the axes and organ damage so as to provide an effective theoretical basis for the development of preventive and treatment measures.

Contagion of depression: a double-edged sword

Depression is a significant mental health issue with extensive economic implications, and recent studies suggest it may be transmitted between individuals. However, the mechanisms of this contagion remain unclear, and the social buffering effect has been understudied. This research employs three rodent models, including stress crossover, cohabitation-induced, and non-contact induced depression contagion models, to explore these mechanisms. Here, we report that that naive mice cohabiting with depressed mice showed increased corticosterone levels and depressive behaviors, unlike those with stressed mice, who did not exhibit these changes and even mitigated desperation in stressed mice. Non-contact cohabitation did not produce significant behavioral differences, but exposure to bedding from depressed mice reduced sucrose preference in naive mice. This study introduces reliable models of depression contagion, suggesting it operates independently of stress transmission. The interplay between depression contagion and social buffering may vary in different contexts. These findings provide new insights into the mechanisms of depression contagion and potential strategies for preventing depressive disorders.

Early positive tactile stimulation reverses the increase of anxiety and decrease of sociality induced by early chronic mechanical pain in mandarin voles

AbstractAnimals may experience early negative (mechanical pain: being retrieved using an incisor by parents or attacked) or positive stimulation (being licked and groomed) that may affect emotional and social behaviors in adulthood. Whether positive tactile stimulation can reverse adverse consequences on emotional and social behaviors in adulthood resulting from chronic mechanical pain and underlying mechanisms remain unclear. This study used a tail‐pinching model during development to simulate mechanical pain experienced by pups in high‐social mandarin voles (Microtus mandarinus). Subsequently, brush‐like positive tactile stimuli were applied to the backs of the mandarin voles. Various behavioral tests were used to measure levels of anxiety, depression, and sociability. The results showed that early tail‐pinching delayed the eye opening of pups, increased levels of anxiety, reduced levels of sociality in male mandarin voles, and impaired social cognition in females during adulthood. Brushing on the back reversed some of these effects. While mandarin voles that were exposed to tail‐pinching during development were exposed to sub‐threshold variable stress as adults, they were more likely to show a stress‐induced increase of anxiety‐like behavior, reduction of sociability, and impairment of social cognition, displaying heightened susceptibility to stress, particularly in males. However, back‐brushing reversed some of these effects, implying that these adults display enhanced stress resilience. In addition, tail‐pinching reduced levels of serum oxytocin and increased corticosterone levels in serum, but back‐brushing reversed these effects. Overall, it was found that positive tactile stimulation reversed increases in anxiety and impairments of social behavior induced by negative stimulation in male mandarin voles via alteration of oxytocin and corticosterone levels.

The effects of Gastrodia elata on anxiety-like behavior in mice

The present study investigated the effect of Gastrodia elata extracts (GE) on chronic electric foot-shock stress (cEFS)-induce anxiety-like behaviors in male C57BL/6 mice. Mice were administered GE (200 or 400 mg/kg/day, oral), and then exposed to cEFS (0.6 mA intensity, 1 s every 5 s interval, 3 min duration) for seven days. GE treatment ameliorates the anxiety-like behavior induced by cEFS, which were determined with elevated plus maze test and open field test. cEFS decreased the dopamine and serotonin levels in brain and both were reversed by GE treatment. cEFS increased corticosterone levels in serum and number of c-fos positive cells in brain, which were downregulated by GE treatment. These results suggested that GE ameliorates the anxiety-like behavior by regulating the levels of dopamine and serotonin, the number of c-fos positive cells in the brain, and corticosterone levels in the serum.

Social stress in rats promotes transcriptional mitochondrial changes of the adrenal tissue

Previous observations suggest that strong social stressors, that is, repeated social defeat, could initiate functional changes in the hypothalamic-pituitary-adrenal (HPA) axis in social mammals. Here, we examine whether exposure to such stress in rats causes persistent alterations in gene expression associated with hypo-and/or hyper-methylation in the adrenal tissue, i.e., the interface between the HPA axis and the circulating hormones. Our integrated analyses of RNA-sequencing (RNAseq) and whole genome bisulfite sequencing (WGBS), suggested that adrenal mitochondrial membrane transport necessary for corticosterone (CORT) synthesis, are affected by repeated social defeat. The increased CORT levels observed in blood in the stressed rats further suggested that corticosterone synthesis might be influenced by this form of social stress. The cellular mechanisms underlying these changes, i.e., the enriched gene ontology (GO)-terms and increased levels of circulating CORT, remain to be investigated.

Time-dependent dual mode of action of COX-2 inhibition on mouse serum corticosterone levels

To elucidate the effect of cyclooxygenase-2 (COX-2) inhibition on corticosterone release, mice were divided into a group receiving NS398, a selective COX-2 inhibitor at a dose of 3 mg/kg for seven days, and a group receiving NS398 for fourteen days. After this time, the mice were sacrificed, and blood serum was collected. An ELISA protocol was used to analyze serum corticosterone levels. Short-term COX-2 inhibition increased corticosterone levels, while long-term inhibition lowered them. The exact schedule of experiments was repeated after the lipopolysaccharide (LPS) Escherichia coli challenge in mice to check the influence of stress stimuli on the tested parameters. In this case, we observed increases in corticosterone levels, significant in a seven-day pattern. These results indicate that corticosterone levels are regulated through a COX-2-dependent mechanism in mice.

Is membrane androgen and estrogen receptor signaling imperative in the governing function of the adrenal cortex in the Eurasian beaver (Castor fiber L.)?

There is a need to fully know the physiology of Eurasian beaver due to its essential role in environmental homeostasis. However, a "human factor" impacts this, including stress conditions and environmental pollution. Adrenal glands protect these all. The regulation of endocrine processes by nonclassical androgen and estrogen signaling, the first and fastest control, is still a matter of research. The specific analyses performed here in mature female and male beaver adrenals contained: anatomical and histological examinations, expression and localization of membrane androgen receptor (zinc transporter, Zinc- and Iron-like protein 9; ZIP9) and membrane estrogen receptor coupled with G protein (GPER), and measurement of zinc (Zn2+) and copper (Ca2+) ion levels and corticosterone levels. We revealed normal anatomical localization, size, and tissue histology in female and male beavers, respectively. Equally,ZIP9 and GPER were localized in the membrane of all adrenal cortex cells. The protein expression of these receptors was higher (p < 0.001) in male than female adrenal cortex cells. Similarly, Zn2+ and Ca2+ ion levels were higher (p < 0.05, p < 0.01) in male than female adrenal cortex. The increased corticosterone levels (p < 0.001) were detected in the adrenal cortex of females when compared to males. The present study is the first to report the presence of nonclassical androgen and estrogen signaling and its possible regulatory function in the adrenal cortex of Eurasian beavers. We assume that this first-activated and fast-transmitted regulation can be important in the context of the effect of environmental physical and chemical stressors especially on adrenal cortex cells. The beaver adrenals may constitute an additional supplementary model for searching for universal mechanisms of adrenal cortex physiology and diseases.

Social stress-induced depressive-like symptoms and changes in gut microbial and lipidomic profiles are prevented by pharmacological inhibition of FAAH activity in male rats

Pharmacological inhibition of fatty acid amide hydrolase (FAAH) activity has antidepressant-like effects in preclinical models of stress. In this study, we investigated whether the antidepressant-like effects of FAAH inhibition are associated with corresponding changes in gut microbial and lipidomic profiles, which are emerging as critical components in the pathophysiology of depression. Adult male Wistar rats experienced five weeks of repeated social defeat or control procedure and were treated with the FAAH inhibitor URB694 (0.3 mg/kg/day, i.p.) or vehicle starting from the third week. Repeated social defeat induced the emergence of depressive-like behavioral (sucrose preference reduction and passive coping behaviors in the forced swim test) and neuroendocrine (increased corticosterone levels) changes, which were prevented by URB694 treatment. Repeated social defeat also provoked a significant variation in gut microbiota (changes in the relative abundance of 14 bacterial taxa) and lipidic (e.g., glycerophospholipids) composition. These stress-induced changes were prevented by URB694 treatment. These findings indicate that inhibition of FAAH activity with URB694 blocks the co-occurrence of depressive-like behavioral and neuroendocrine changes and alterations in gut microbial and lipid composition in rats exposed to repeated social defeat. In conclusion, these results suggest that the gut microbiota-lipid crosstalk may represent a novel biological target for FAAH inhibitors to enhance stress resilience.

Vagus nerve damage increases alcohol intake and preference in a nonpreferring rat line: Relationship to vagal regulation of the hypothalamic-pituitary-adrenal axis.

Clinical and preclinical research indicates that gastric weight loss surgeries, such as Roux-en-Y gastric bypass surgery, can induce alcohol use disorder (AUD). While numerous mechanisms have been proposed for these effects, one relatively unexplored potential mechanism is physical damage to the gastric branch of the vagus nerve, which can occur during bypass surgery. Therefore, we hypothesized that direct damage to the gastric branch of the vagus nerve, without altering other aspects of gastric anatomy, could result in increased alcohol intake. To test this hypothesis, we compared alcohol intake and preference in multiple models in male Sprague-Dawley rats that received selective gastric branch vagotomy (VX) with rats who underwent sham surgery. Because the vagus nerve regulates hypothalamic-pituitary-adrenal (HPA) axis function, and alterations to HPA function are critical to the escalation of non-dependent alcohol intake, we also tested the hypothesis that gastric VX increases HPA function. We found that VX increases alcohol intake and preference in the every-other-day, two-bottle choice test and increases preference for 1 g/kg alcohol in the conditioned place preference test. The effects were selective for alcohol, as sucrose intake and preference were not altered by VX. We also found that VX increases corticotropin releasing factor (CRF) mRNA in the paraventricular nucleus of the hypothalamus (PVN), increases putative PVN CRF neuronal action potential firing, and increases corticosterone levels. Overall, these findings suggest that the vagus nerve may play a critical role in regulating HPA axis function via modulation of PVN CRF mRNA expression and putative PVN CRF neuronal activity. Furthermore, disruptions to vagal regulation of HPA axis function may increase alcohol intake and preference.

ВЛИЯНИЕ ЭКСПЕРИМЕНТАЛЬНОЙ ДЕПРЕССИИ НА ФУНКЦИОНАЛЬНОЕ СОСТОЯНИЕ ГИПОТАЛАМО-ГИПОФИЗАРНО-НАДПОЧЕЧНИКОВОЙ СИСТЕМЫ

Objective: To study the effect of modelled depression not associated with stress on the hypothalamic-pituitary-adrenal system (HPAS) in mice. Methods: The experiment was conducted on the twenty-four C57BL/6 mice weighing 20-25 g, divided into two groups: an experimental one, involving animals with reserpine-induced depression, and a control one, the animals of which were administered normal saline instead of reserpine solution. At the end of the experiment, behavioral tests were performed (open field, sucrose preference, forced swimming), and the corticosterone concentration in blood was measured using ELISA. Image analysis of histological changes in the adrenal gland and thymus was performed. All results were statistically processed using SPSS 27.0.1 software, with p&lt;0.05 indicating the significance of differences. Results: Animals of the experimental group showed a significantly higher level of corticosterone in the blood (p&lt;0.01), significant changes in behavior in the open field test (distance traveled, rearing, frequency of urination and defecation, p&lt;0.01), reduced sucrose preference (p&lt;0.01) and shortened duration of the active phase in the forced swimming test (p&lt;0.01) than the animals of the control group. Besides, in the mice of the experimental group, a significant increase in the volume density of the zona fasciculata of the adrenal cortex was found (p&lt;0.05), as well as the area of its spongiocytes and their nuclei (p&lt;0.05). In the thymus of the experimental animals, a significant decrease in the cortex-medulla ratio was noted (p&lt;0.05); and an increase in the volume density of tingible body macrophages (p&lt;0.05) compared to the control group of mice. Conclusion: Modelled depression, not associated with stress, the depth of which was assessed by behavioral tests, caused activation of the HPAS, manifested by increased corticosterone levels, hyperplasia of the zona fasciculata of the adrenal cortex, hypertrophy of its spongiocytes and their nuclei, as well as changes in the thymus, indicating associated immunosuppression. Thus, the study showed that, along with the well-known phenomenon of the development of depression against the background of chronic stress-associated activation of HPAS, there is an inverse mechanism leading to activation of the HPAS during the development of depression not associated with stress, which must be taken into account when developing the new strategies of treating depression. Keywords: Depression, hypothalamic-pituitary-adrenal system, corticosterone, immune system, mice.

Outcomes of early social experiences on glucocorticoid and endocannabinoid systems in the prefrontal cortex of male and female adolescent rats.

Social and emotional experiences differently shape individual's neurodevelopment inducing substantial changes in neurobiological substrates and behavior, particularly when they occur early in life. In this scenario, the present study was aimed at (i) investigating the impact of early social environments on emotional reactivity of adolescent male and female rats and (ii) uncovering the underlying molecular features, focusing on the cortical endocannabinoid (eCB) and glucocorticoid systems. To this aim, we applied a protocol of environmental manipulation based on early postnatal socially enriched or impoverished conditions. Social enrichment was realized through communal nesting (CN). Conversely, an early social isolation (ESI) protocol was applied (post-natal days 14-21) to mimic an adverse early social environment. The two forms of social manipulation resulted in specific behavioral and molecular outcomes in both male and female rat offspring. Despite the combination of CN and ESI did not affect emotional reactivity in both sexes, the molecular results reveal that the preventive exposure to CN differently altered mRNA and protein expression of the main components of the glucocorticoid and eCB systems in male and female rats. In particular, adolescent females exposed to the combination of CN and ESI showed increased corticosterone levels, unaltered genomic glucocorticoid receptor, reduced cannabinoid receptor type-1 and fatty acid amide hydrolase protein levels, suggesting that the CN condition evokes different reorganization of these systems in males and females.

Acute restraint stress regulates brain DNMT3a and promotes defensive behaviors in male rats

Depending on its duration and severity, stress may contribute to neuropsychiatric diseases such as depression and anxiety. Studies have shown that stress impacts the hypothalamic–pituitary–adrenal (HPA) axis, but its downstream molecular, behavioral, and nociceptive effects remain unclear. We hypothesized that a 2-hour single exposure to acute restraint stress (ARS) activates the HPA axis and changes DNA methylation, a molecular mechanism involved in the machinery of stress regulation. We further hypothesized that ARS induces anxiety-like and risk assessment behavior and alters nociceptive responses in the rat. We employed biochemical (radioimmunoassay for corticosterone; global DNA methylation by enzyme immunoassay and western blot for DNMT3a expression in the amygdala, ventral hippocampus, and prefrontal cortex) and behavioral (elevated plus maze and dark-light box for anxiety and hot plate test for nociception) tests in adult male Wistar rats exposed to ARS or handling (control). All analyses were performed 24 h after ARS or handling. We found that ARS increased corticosterone levels in the blood, increased the expression of DNMT3a in the prefrontal cortex, promoted anxiety-like and risk assessment behaviors in the elevated plus maze, and increased the nociceptive threshold observed in the hot plate test. Our findings suggest that ARS might be a helpful rat model for studying acute stress and its effects on physiology, epigenetic machinery, and behavior.

Not so fast: Paradoxically increased variability in the glucose tolerance test due to food withdrawal in continuous glucose-monitored mice

ObjectiveThis study was performed to determine the effect of fasting on reproducibility of the glucose tolerance test. Due to individual variation in animal feeding behaviors, fasting animals prior to metabolic and behavioral experiments is widely held to reduce inter-subject variation in glucose and metabolic parameters of preclinical rodent models. Reducing variability is especially important for studies where initial metabolite levels can influence the magnitude of experimental interventions, but fasting also imposes stress that may distort the variables of interest. One such intervention is the glucose tolerance test (GTT) which measures the maximum response and recovery following a bolus of exogenous glucose. We sought to investigate how fasting affects the response of individual mice to a GTT. MethodsUsing simultaneous continuous glucose monitoring (CGM) and indirect calorimetry, we quantified blood glucose, physical activity, body temperature, metabolic rates, and food consumption levels on a minute-to-minute basis in adult male mice for 4 weeks. We tested the effects of a 4-h or 18-h fast on the GTT to examine the effect of food withdrawal in light or dark photoperiods. Studies were also performed with 4-h fasting in additional mice without implanted CGM probes. ResultsContrary to our expectations, a 4-h fast during the light photoperiod promotes a paradoxical increase in inter-animal variation in metabolic rate, physical activity, body temperature, glycemia, and glucose tolerance. This hyperglycemic and hyper-metabolic phenotype promotes increased corticosterone levels and is consistent with a behavioral stress response to food deprivation, even in well-fed mice. We find that mice undergoing an 18-h fast entered torpor, a hibernation-like state. In addition to low body temperature and metabolic rate, torpor is also associated with glucose levels 56 mg/dl lower than those seen in mice with ad libitum access to food. Moreover, the time spent in torpor affects the response to a GTT. ConclusionOur results suggest fasting mice before glucose tolerance testing, and perhaps other experiments, can have the opposite of the intended effect where fasting can increase, rather than decrease, experimental variability.

Conservation implications of tourism and stress for Amazonian caimans

AbstractEcotourism is a strategy for biodiversity conservation, but it involves possible negative effects on animal health and welfare. Large predators such as crocodilians are one of the great public attractions sought after for tourist interactions. Interactions with wild animals and humans can hyperstimulate the hypothalamic‐pituitary‐adrenal axis in the short term, in the case of crocodilians showing an increased corticosterone level, which is indicative of physiological stress. Between September and December 2019, we simulated interactions between tourists and Amazonian crocodilians at Anavilhanas National Park in Central Amazonia, Brazil to evaluate the effects of handling and use of photographic flashes on black caiman (Melanosuchus niger) and spectacled caiman (Caiman crocodilus) on circulating corticosterone and lactate. Corticosterone levels increased 1.7‐fold during handling and 2.7‐fold when exposed to photographic flashes in black caiman but not in spectacled caiman. Increased corticosterone concentrations in black caiman were characterized by an increase caused by handling and were more intense after flashes than in controls, but the combination of handling and flash had no effect. During handling in simulated tourist interactions, anaerobic respiration increased lactate in black caiman but not in spectacled caiman. The effect of simulated tourist interactions with Amazonian crocodilians was dependent on the handling and especially on flash use in black caiman. The results can assist management, conservation programs, and public policies, especially in programs based on tourism interaction with Amazonian crocodilians.

Repeated neonatal isoflurane exposure facilitated stress-related fear extinction impairment in male mice and was associated with ΔFosB accumulation in the basolateral amygdala and the hippocampal dentate gyrus

Volatile anesthetics elicit neurodevelopmental toxicity in rodents and primates and lead to more exaggerated anxiety-like behavior in response to future stress. Anxiety and fear are closely correlated and maladaptive fear-associated learning is regarded as the core mechanism underlying anxiety-related disorders. However, little is known about the interaction between early-life anesthetic exposure and future stress and the accompanying effect on fear-associated learning. In the present study, we evaluated the alterations in fear-associated learning (fear acquisition and extinction) occurring in mice receiving repeated neonatal isoflurane exposure and chronic variable stress (CVS) successively through a series of fear conditioning, fear reinforcing, and fear extinction paradigms. The corticosterone (CORT) response during CVS and the immunohistochemical levels of ΔFosB and c-Fos expression in the basolateral amygdala (BLA) and the hippocampal dentate gyrus (DG) after the extinction retrieval test were also investigated. The results showed that neonatal isoflurane exposure could increase CORT levels following the first diurnal CVS procedure, but not after completion of the whole CVS paradigm. Neonatal isoflurane exposure exerted a repressive effect on fear acquisition, in contrast to that seen with CVS. Neonatal isoflurane exposure and CVS both exerted suppressive effects on fear extinction and there was a significant synergy between them. Furthermore, neonatal isoflurane exposure facilitated CVS-mediated ΔFosB accumulation in the BLA and the hippocampal DG, which may have been responsible for c-Fos expression deficits and fear extinction impairment. Collectively, these findings contribute to the understanding of the interaction between early-life anesthetic exposure and future stress, as well as the accompanying behavioral alterations.