Differences In Corticosterone Levels Research Articles

Nicotine exposure through breastfeeding affects brain-derived neurotrophic factor and synaptic proteins levels in the brain of stressed adult female mice.

Nicotine has been used during pregnancy and lactation as a tobacco harm reduction strategy. However, it is unclear whether nicotine exposure during a critical development period negatively impacts stress responses in adulthood. This study investigated how nicotine, administered via breastfeeding, affects the brain-derived neurotrophic factor (BDNF), synaptic proteins levels, and anxiety-like behavior in adult female mice subjected to stress. Female Swiss mice were exposed to saline or nicotine (8mg/kg/day) through breastfeeding between their fourth and 17th postnatal days (P) via implanted osmotic mini pumps. The unpredictable chronic mild stress (UCMS) protocol was performed during their adulthood (P65) for 10 consecutive days, followed by the elevated plus maze (EPM) test 1 day after the protocol. Animals were euthanized and their blood, collected for plasma corticosterone measurements and their brain structures, dissected for BDNF and synaptic proteins analyses. We found no significant differences in corticosterone levels between groups (Saline/Non-stress, Nicotine/Non-stress, Saline/Stress, and Nicotine/Stress). The UCMS protocol hindered weight gain. Mice exposed to nicotine through breastfeeding with or without the UCMS protocol in adulthood showed higher grooming and head dipping frequency; decreased BDNF levels in cerebellum and striatum; increased postsynaptic density protein 95 (PSD-95), synapsin I, and synaptophysin levels in cerebellum; and decreased PSD-95 and synapsin I levels in brainstem. Our results indicate that nicotine exposure through breastfeeding leads to long-lasting behavioral effects and synaptic protein changes, most of which were independent of the UCMS protocol, even after a long nicotine-free period, highlighting the importance of further studies on nicotine exposure during development.

Impact of Refinements to Handling and Restraint Methods in Mice.

Simple SummaryFor laboratory mice, handling methods used for routine husbandry and procedures can cause stress, particularly if tail handling methods are used. We first sought to investigate if there were any measurable impacts of different handling methods when tested in a controlled real-world study. Animals were handled at our breeding site using different methods, and their impacts on handling post-transport to the research site were analysed by animal care staff. We found a clear effect of handling method on technician scores relating to overt signs of stress and how easy they were to handle. Based on this study, the institution implemented a refined mouse handling policy. However, one barrier to the use of non-tail handling methods has been arguments that tail handling is necessary for restraint in mice. With a relatively minor adaptation to the conventional restraint method, we have been able to combine cupping with physical restraint for procedures avoiding any use of the tail. Here we provide data showing that our modified restraint method is associated with reduced signs of aversion in the mice. Together, our findings support the implementation of refined handling policies for mice, and this can include handling for both husbandry and restraint for procedures.There is increasing evidence that, compared to non-aversive handling methods (i.e., tunnel and cupping), tail handling has a negative impact on mouse welfare. Despite this evidence, there are still research organisations that continue to use tail handling. Here, we investigated handling for routine husbandry by three different methods: tail, cupping and tube in a relevant real-world scenario involving mice bred off-site. After transfer to the destination unit, mice were assessed for overt behaviours associated with anxiety and fear. Mice that experienced tail handling were less easy to handle, were more responsive to the box opening, and scored lower in a hand approach test. One barrier to non-tail handling methods is the current practice of restraining mice by the tail for procedures. We therefore next assessed whether a modified method for restraint that takes the animal from cupping to restraint without the use of the tail was associated with better welfare. This refined restraint method reduced overt signs of distress although we did not find any differences in corticosterone levels or anxiety-related behaviours. These findings suggest that avoiding tail handling throughout the animal’s laboratory experience, including during restraint, benefits their welfare.

Sex-specific effects of hatching order on nestling baseline corticosterone in a wild songbird

Variation in nestling growth and survival is often influenced by hatching order, with first-hatched offspring having an advantage over later-hatched younger siblings. In house wrens (Troglodytes aedon), this effect of hatching order is especially evident in asynchronously hatched broods and can lead to sex-specific differences in the size and condition of nestlings. Females appear to allocate the sex of their offspring across the laying order to capitalize on these differences. We hypothesized that levels of circulating corticosterone, the primary metabolic hormone in birds, mediates these sex-specific effects in nestlings. We predicted that: i) baseline levels of corticosterone in nestlings should vary along the hatching order, ii) effects of hatching order on baseline corticosterone should be sex specific, and iii) any sex-specificity of hatching order on baseline corticosterone could be contingent on the degree of hatching synchrony. We tested these predictions in a study in which we measured baseline corticosterone in first- and last-hatched nestlings in synchronously and asynchronously hatching broods. To assess whether any differences in nestling baseline corticosterone levels could be attributed to pre-natal maternal effects, the post-natal environment, or both, we conducted two additional studies in which we measured i) yolk corticosterone in first- and last-laid eggs and ii) baseline corticosterone in nestlings that were cross-fostered to create simulated ‘asynchronously’ hatched broods. There was a significant interaction between sex and relative hatching order in their effects on nestling baseline corticosterone, but no effect of hatching synchrony. Corticosterone levels remained relatively constant across the hatching order in males but decreased in females. There was a significant effect of laying order on yolk corticosterone, with first-laid eggs containing significantly higher levels of yolk corticosterone than last-laid eggs. Cross-fostering of nestlings at different points of development had no significant effect on nestling corticosterone levels. These results indicate that sex-dependent differences in corticosterone levels across the hatching order may arise, at least in part, from embryonic exposure to maternally derived corticosterone, whereas the post-natal rearing environment plays, at best, a minimal role in determining nestling baseline corticosterone levels.

Does a Red House Affect Rhythms in Mice with a Corrupted Circadian System?

The circadian rhythms of body functions in mammals are controlled by the circadian system. The suprachiasmatic nucleus (SCN) in the hypothalamus orchestrates subordinate oscillators. Time information is conveyed from the retina to the SCN to coordinate an organism’s physiology and behavior with the light/dark cycle. At the cellular level, molecular clockwork composed of interlocked transcriptional/translational feedback loops of clock genes drives rhythmic gene expression. Mice with targeted deletion of the essential clock gene Bmal1 (Bmal1−/−) have an impaired light input pathway into the circadian system and show a loss of circadian rhythms. The red house (RH) is an animal welfare measure widely used for rodents as a hiding place. Red plastic provides light at a low irradiance and long wavelength—conditions which affect the circadian system. It is not known yet whether the RH affects rhythmic behavior in mice with a corrupted circadian system. Here, we analyzed whether the RH affects spontaneous locomotor activity in Bmal1−/− mice under standard laboratory light conditions. In addition, mPER1- and p-ERK-immunoreactions, as markers for rhythmic SCN neuronal activity, and day/night plasma corticosterone levels were evaluated. Our findings indicate that application of the RH to Bmal1−/− abolishes rhythmic locomotor behavior and dampens rhythmic SCN neuronal activity. However, RH had no effect on the day/night difference in corticosterone levels.

Vendor differences in anxiety-like behaviors in female and male Sprague Dawley rats

Although Sprague Dawley outbred rats are commonly used in behavioral, physiological, and pharmacological studies, dramatic differences in responses may emerge from rats obtained from different suppliers even when sex, age, and environmental conditions are maintained constant. In the present study, we compared behavioral responses on three tests related to anxiety of Sprague Dawley female and male rats obtained from three different vendors in the United States: Charles River, Envigo, and Taconic. All rats were tested in the open field, light-dark box, and elevated zero maze. We found reduced time spent in the center area of the open field and decreased light compartment duration in the light-dark box test in female and male rats from Taconic compared to Charles River and Envigo rats, suggesting anxiety-like behaviors differ between the three vendors. No vendor differences were found on performance in the elevated zero maze. Furthermore, the contribution of stress hormones to vendor differences was examined by measuring serum corticosterone levels in rats 30 min after exposure to the elevated zero maze. There were no vendor differences in corticosterone levels, suggesting that endogenous levels of stress hormones most likely did not contribute to vendor differences in anxiety-like behaviors. Collectively, these findings highlight the importance of vendor selection of the Sprague Dawley stock for research involving behavioral tests related to anxiety.

Habitat Characteristics in Created Vernal Pools Impact Spotted Salamander Water-Borne Corticosterone Levels

Spotted salamanders (Ambystoma maculatum) require vernal pools for breeding habitat. Limited protection and preservation of vernal pools makes suitable habitat creation important. Differences in corticosterone levels, a hormone associated with growth, development, and stress in amphibians, could indicate population health and habitat quality. Our objective was to determine if habitat characteristics in created vernal pools influence corticosterone levels of spotted salamander larvae. In May and June of 2015 and 2016, we sampled water-borne corticosterone levels of larval spotted salamanders in 34 created vernal pools constructed 1–5 years earlier. Using multiple regression, we determined the best model predicting corticosterone levels included larval total length, pool-water temperature, year sampled, and pool diameter. Pool-water pH, depth, and age; percent cover; and predator presence were not significant predictors. Annual variation in corticosterone levels and habitat characteristics, and positive associations with water temperature and salamander body size highlighted the importance of controlling for external influences. The negative association between pool diameter and corticosterone indicated that larvae in larger pools (up to 12.75-m maximum diameter) were less stressed and potentially healthier. These results indicate that pool diameter contributes to habitat quality and may be important when constructing vernal pools for spotted salamanders.

Caloric restriction in grouped rats: aggregate influence on behavioural and hormonal data.

The majority of studies in short- and middle-term caloric restriction (CR) have been primarily focused on physiological parameters, improvements in aging, modulation of oxidative stress, and long-term negative effects on cognitive functions. However, single-housing associated with CR may pose many logistical problems. Thus, it is necessary to study the effects of CR under conditions in which animals are group-housed. The aims of this study were to (i) observe the possible differences in the proportion of the weights and social behaviour under ad libitum and CR (at 70%) conditions; (ii) examine the eventual inequalities in the proportion of the weights and social behaviour (the time spent eating under the feeder as an indicator of dominance and empathy, and the number of 'pushes' as an indicator of aggressiveness) in sibling and non-sibling rats under CR conditions; and (iii) compare the concentrations of corticosterone (stress biomarker) in serum under ad libitum and CR conditions. The results indicated the effectiveness of CR in different groups independent of the relationship between the rats. No extreme changes in weight were observed in the CR rats. Behavioural observations also indicated the differences in the total time spent under the feeder and in the number of pushes (higher in both cases for the sibling rats). However, no significant differences in corticosterone levels were observed. Our results suggest the viability of group-housing rats during long periods of CR maintenance.

Altered corticosterone levels and social play behavior after prolonged maternal separation in adolescent male but not female Wistar rats

Early-life socio-environmental factors are crucial for normal developmental processes; adverse experiences early in life can therefore lead to detrimental effects in several physiological systems. The aim of this study was to examine short-term effects of early adverse experiences in a maternal separation (MS) rodent model. In this study two separation conditions were used: daily 15- (MS15) or 360-min (MS360) separation of the litter from the dam during postnatal day 1–21. In early adolescence, male and female offspring were subjected to a single-isolation procedure with analysis of corticosterone levels prior to and after isolation. In addition, social play behavior was assessed during mid-adolescence. There was a clear difference between male and female offspring in both tests performed. There was no difference in corticosterone levels between the female MS groups, whereas MS360 males showed higher baseline and recovery corticosterone levels than MS15 males. The amount of pinning, a specific social play behavior, was affected by rearing with MS360 males having a higher frequency than MS15 males, while there was no difference between the female MS groups. The observation that males but not females are affected by MS360 has previously been reported for adult animals, and herein we show that this difference is present already in adolescence. Changes in corticosterone levels and social behavior following early-life adversity have been associated with adult behavioral alterations, and our results confirm that these changes emerge already within adolescence.

The effect of light level, CO2 flow rate, and anesthesia on the stress response of mice during CO2 euthanasia.

Euthanasia protocols are designed to mitigate the stress experienced by animals, and an environment that induces minimal stress helps achieve that goal. A protocol that is efficient and practical in a typical animal research facility is also important. Light intensity, isoflurane, and CO2 flow rate were studied for their impact on the stress response of mice during CO2 euthanasia. Behavior was observed and scored during euthanasia and serum corticosterone was measured immediately after death. Unsurprisingly, animals euthanized with a high-flow rate of CO2 became unconscious in the least amount of time, while animals euthanized with a low-flow rate required the most time to reach unconsciousness. There was a significant increase in anxious behaviors in animals in the isoflurane group (F1,12 = 6.67, P = 0.024), the high-flow rate CO2 group (F1,12 = 10.24, P = 0.007), and bright chamber group (F1,12 = 7.27, P = 0.019). Serum corticosterone was highest in the isoflurane group (124.72 ± 83.98 ng/ml), however there was no significant difference in corticosterone levels observed for the other study variables of light and flow-rate. A darkened chamber and low CO2 flow rates help to decrease stress experienced during CO2 euthanasia, while the use of isoflurane was observed to increase the stress response during euthanasia.

Effect of Repeated 1-h Episodes of Immobilization Stress on Activity of Glucocorticoid Metabolism Enzymes in the Liver.

Differences in corticosterone level associated with different activity of glucocorticoid-oxidizing enzymes in the liver were revealed in rats exposed to stress. Pro-inflammatory changes in the liver were associated with enhanced CYP3A-dependent monooxygenation.

Intermittent fasting favored the resolution of Salmonella typhimurium infection in middle-aged BALB/c mice.

Intermittent fasting (IF) reportedly increases resistance and intestinal IgA response to Salmonella typhimurium infection in mature mice. The aim of this study was to explore the effect of aging on the aforementioned improved immune response found with IF. Middle-aged male BALB/c mice were submitted to IF or ad libitum (AL) feeding for 40weeks and then orally infected with S. typhimurium. Thereafter, infected animals were all fed AL (to maximize their viability) until sacrifice on day 7 or 14 post-infection. We evaluated body weight, bacterial load (in feces, Peyer's patches, spleen and liver), total and specific intestinal IgA, lamina propria IgA+ plasma cells, plasma corticosterone, and messenger RNA (mRNA) expression of α-chain, J-chain, and the polymeric immunoglobulin receptor (pIgR) in liver and intestinal mucosa. In comparison with the infected AL counterpart, the infected IF group (long-term IF followed by post-infection AL feeding) generally had lower intestinal and systemic bacterial loads as well as higher total IgA on both post-infection days. Both infected groups showed no differences in corticosterone levels, body weight, or food and caloric intake. The increase in intestinal IgA was associated with enhanced pIgR mRNA expression in the intestine (day 7) and liver. Thus, to maintain body weight and caloric intake, IF elicited metabolic signals that possibly induced the increased hepatic and intestinal pIgR mRNA expression found. The increase in IgA probably resulted from intestinal IgA transcytosis via pIgR. This IgA response along with phagocyte-induced killing of bacteria in systemic organs (not measured) may explain the resolution of the S. typhimurium infection.

Early rearing experience is associated with vasopressin immunoreactivity but not reactivity to an acute non-social stressor in the prairie vole

The early life experiences of an organism have the potential to alter its developmental trajectories. Perhaps one of the most powerful influences during this period is the parent–offspring relationship. Previous work in several mammalian species has demonstrated that parental care in early life and specifically maternal behavior can influence several adult outcomes in offspring, including affiliative and aggressive behavior, parental behavior, hypothalamic–pituitary–adrenal (HPA) functioning and risk of psychopathology. We have previously demonstrated that naturally occurring variation in the type and amount of care given to offspring in a biparental species, the prairie vole (Microtus ochrogaster), is related to social, anxiety-like, aggressive behaviors as well as HPA response to chronic and acute social stressors. Here we aim to determine the effects of early biparental care on HPA functioning and the interaction between early care and later reactivity to a forced swim test, an acute non-social stressor. Behavior during the swim test as well as several indicators of HPA activity, including plasma corticosterone (CORT), corticotropin releasing hormone immunoreactivity (CRH-ir), and vasopressin immunoreactivity (AVP-ir) were measured. Results here indicate an effect of early experience on AVP-ir but not CRH-ir or plasma CORT. There were no differences in CORT levels between high-contact (HC) and low-contact (LC) males or females for either control animals or after a swim stressor. CRH-ir was higher in the central amygdala following a swim test but was not influenced by early care. However, AVP-ir was not influenced by exposure to a swim stressor but was affected by early parental care in a sex-dependent manner. Female HC offspring had increased AVP-ir in the SON while HC male offspring had decreased AVP-ir in the PVN compared to their LC counterparts. The differential response of CRH and AVP to early experience and later stress, and the lack of an interaction between early care rearing and later adult stress, suggest an independence in response of some components of the HPA system. In addition, these findings expand our understanding of the relationship between naturally occurring variation in early biparental care and sexual dimorphisms in adult outcomes.

Experimental food restriction reveals individual differences in corticosterone reaction norms with no oxidative costs.

Highly plastic endocrine traits are thought to play a central role in allowing organisms to respond rapidly to environmental change. Yet, not all individuals display the same degree of plasticity in these traits, and the costs of this individual variation in plasticity are unknown. We studied individual differences in corticosterone levels under varying conditions to test whether there are consistent individual differences in (1) baseline corticosterone levels; (2) plasticity in the hormonal response to an ecologically relevant stressor (food restriction); and (3) whether individual differences in plasticity are related to fitness costs, as estimated by oxidative stress levels. We took 25 wild-caught house sparrows into captivity and assigned them to repeated food restricted and control treatments (60% and 110% of their daily food intake), such that each individual experienced both food restricted and control diets twice. We found significant individual variation in baseline corticosterone levels and stress responsiveness, even after controlling for changes in body mass. However, these individual differences in hormonal responsiveness were not related to measures of oxidative stress. These results have implications for how corticosterone levels may evolve in natural populations and raise questions about what we can conclude from phenotypic correlations between hormone levels and fitness measures.

Bifidobacteria exert strain-specific effects on stress-related behavior and physiology in BALB/c mice.

Accumulating evidence suggests that commensal bacteria consumption has the potential to have a positive impact on stress-related psychiatric disorders. However, the specific bacteria influencing behaviors related to anxiety and depression remain unclear. To this end, we compared the effects of two different Bifidobacteria on anxiety and depression-like behavior; an antidepressant was also used as a comparator. Innately anxious BALB/c mice received daily Bifidobacterium longum (B.) 1714, B. breve 1205, the antidepressant escitalopram or vehicle treatment for 6 weeks. Behavior was assessed in stress-induced hyperthermia test, marble burying, elevated plus maze, open field, tail suspension test, and forced swim test. Physiological responses to acute stress were also assessed. Both Bifidobacteria and escitalopram reduced anxiety in the marble burying test; however, only B. longum 1714 decreased stress-induced hyperthermia. B. breve 1205 induced lower anxiety in the elevated plus maze whereas B. longum 1714 induced antidepressant-like behavior in the tail suspension test. However, there was no difference in corticosterone levels between groups. These data show that these two Bifidobacteria strains reduced anxiety in an anxious mouse strain. These results also suggest that each bacterial strain has intrinsic effects and may be beneficially specific for a given disorder. These findings strengthen the role of gut microbiota supplementation as psychobiotic-based strategies for stress-related brain-gut axis disorders, opening new avenues in the field of neurogastroenterology.

The effect of the Life Enhancement Treatment for Substance Use (LETS ACT) on cocaine relapse: Results from a randomized control trial

The effect of the Life Enhancement Treatment for Substance Use (LETS ACT) on cocaine relapse: Results from a randomized control trial

Sex-specific phenotypic integration: endocrine profiles, coloration, and behavior in fledgling boobies

The intensity of color expression in animals plays a key role in social environments as a mechanism to signal individual capacities in competitive contests. Selective pressures for resource competition differ at different stages of life and between sexes; therefore, coloration is expected to vary between juveniles and adults and between males and females. Exploring the covariance between coloration and other traits may help to understand the functional significance of color and the action of natural selection on multivariate phenotypes. Melanin-based plumage coloration was investigated in the masked booby Sula dactylatra in relation to melanin concentration, sex, hormone levels, and shy–bold behavior of chicks close to fledging. Darker brown boobies showed higher levels of both eumelanin and pheomelanin concentration and lower body mass. Males behaved bolder than females and showed on average 8% larger brown patches. Bolder females had smaller brown patches. Bolder individuals also had lower levels of circulating testosterone, but no differences in corticosterone levels were found. Stronger phenotypic integration was observed in females than males. Our study suggests that juvenile melanic coloration may reflect behavioral strategies by sex, endocrine profiles, and body mass indicating the convergence of different adaptive functions in a given phenotype, this being more evident in females. Direction of correlations differed from those predicted under the pleiotropic idea for color-related traits. These results suggest the possibility that juvenile plumage acts as a signaling system in a social context within the age class and suggest that plumage coloration may indicate different behavioral strategies.

A flexible Bayesian hierarchical approach for analyzing spatial and temporal variation in the fecal corticosterone levels in birds when there is imperfect knowledge of individual identity

Population cycles have long interested biologists. The ruffed grouse, Bonasa umbellus, is one such species whose populations cycle over most of their range. Thus, much effort has been expended to understand the mechanisms that might control cycles in this and other species. Corticosterone metabolites are widely used in studies of animals to measure physiological stress. We evaluated corticosterone metabolites in feces of territorial male grouse as a potential tool to study mechanisms governing grouse cycles. However, like most studies of corticosterone in wild animals, we did not know the identity of all individuals for which we had fecal samples. This presented an analytical problem that resulted in either pseudoreplication or confounding. Therefore, we derived an analytical approach that accommodated for uncertainty in individual identification. Because we had relatively low success capturing birds, we estimated turnover probabilities of birds on territorial display sites based on capture histories of a limited number of birds we captured. Hence, we developed a study design and modeling approach to quantify variation in corticosterone levels among individuals and through time that would be applicable to any field study of corticosterone in wild animals. Specifically, we wanted a sampling design and model that was flexible enough to partition variation among individuals, spatial units, and years, while incorporating environmental covariates that would represent potential mechanisms. We conducted our study during the decline phase of the grouse cycle and found high variation among corticosterone samples (11.33–443.92ng/g [x¯=113.99ng/g, SD=69.08, median=99.03ng/g]). However, there were relatively small differences in corticosterone levels among years, but levels declined throughout each breeding season, which was opposite our predictions for stress hormones correlating with a declining population. We partitioned the residual variation into site, bird, and repetition (i.e., multiple samples collected from the same bird on the same day). After accounting for years and three general periods within breeding seasons, 42% of the residual variation among observations was attributable to differences among individual birds. Thus, we attribute little influence of site on stress level of birds in our study, but disentangling individual from site effects is difficult because site and bird are confounded. Our model structures provided analytical approaches for studying species having different ecologies. Our approach also demonstrates that even incomplete information on individual identity of birds within samples is useful for analyzing these types of data.

Effects of aging on hypothalamic-pituitary-adrenal (HPA) axis activity and reactivity in virgin male and female California mice (Peromyscus californicus)

Life history theory posits that organisms face a trade-off between current and future reproductive attempts. The physiological mechanisms mediating such trade-offs are still largely unknown, but glucocorticoid hormones are likely candidates as elevated, post-stress glucocorticoid levels have been shown to suppress both reproductive physiology and reproductive behavior. Aged individuals have a decreasing window in which to reproduce, and are thus predicted to invest more heavily in current as opposed to future reproduction. Therefore, if glucocorticoids are important in mediating the trade-off between current and future reproduction, aged animals are expected to show decreased hypothalamic-pituitary-adrenal (HPA) axis responses to stressors and to stimulation by corticotropin-releasing hormone (CRH), and enhanced responses to glucocorticoid negative feedback, as compared to younger animals. We tested this hypothesis in the monogamous, biparental California mouse by comparing baseline and post-stress corticosterone levels, as well as corticosterone responses to dexamethasone (DEX) and CRH injections, between old (∼18–20months) and young (∼4months) virgin adults of both sexes. We also measured gonadal and uterine masses as a proxy for investment in potential current reproductive effort. Adrenal glands were weighed to determine if older animal had decreased adrenal mass. Old male mice had lower plasma corticosterone levels 8h after DEX injection than did young male mice, suggesting that the anterior pituitary of older males is more sensitive to DEX-induced negative feedback. Old female mice had higher body-mass-corrected uterine mass than did young females. No other differences in corticosterone levels or organ masses were found between age groups within either sex. In conclusion, we did not find strong evidence for age-related change in HPA activity or reactivity in virgin adult male or female California mice; however, future studies investigating HPA activity and reproductive outcomes in young and old breeding adults would be illuminating.

2014 – The role of adenylyl cyclase 8 in stress response

Adelylyl cyclase 8(AC8) is a brain specific cyclase activated only by calcium. Therefore its activity has been associated with plasticity process underlying learning and memory. AC8 knockout(KO) mice show impaired stress-induced anxiety. Here we try to find whether AC8 is involved in hypothalamus-hypophysis-axis(HPA) regulation and if it influences intracellular signaling of gluccocorticoids in the hippocampus. First, serum samples were harvest from 8 AC8 KO and 8 wild type(WT) mice at basal, eve, 30 and 120 minutes after stress and at sacrifice with subsequent measuring of corticosterone levels. Second, glucocorticoid receptor(GR), vasopressin(AVP) and corticotrophin releasing hormone(CRH) mRNA expression levels were measured in accumbens nucleus(Acb), periventricular nucleus(PVN), supraoptic nucleus(SON), amygdala and hippocampus. Finally, GR and phosphorilated-GR levels were measured in hippocampus by western blot technique. There was no difference in corticosterone levels between the two groups, regardless of harvesting time (p > 0.05). Also, we found no difference between AVP mRNA levels in PVN or SON (p > 0.05). However, AC8KO mice tend to have lower CRH levels in amygdala and lower GR levels in the hippocampus (p < 0.05). More, AC8KO mice have higher phosphorilated-GR levels in hippocampus in comparison with WT (p = 0.01). Our data shoes that AC8 does not play a part in HPA regulation. However, it alters the expression levels and phosphorilation patterns of GR in the hippocampus, which can explain the impaired learned anxiety behavior. Since anxiety-impaired stress response is one of few outcomes of AC8 KO phenotype, we imply that this cyclase might represent a potential target for specific antianxiety drugs.

Chronic adrenocorticotrophic hormone treatment alters tricyclic antidepressant efficacy and prefrontal monoamine tissue levels

Several animal models are currently utilised in the investigation of major depressive disorder; however, each is validated by its response to antidepressant pharmacotherapy. Few animal models consider the notion of antidepressant treatment resistance. Chronic daily administration of adrenocorticotropic hormone (ACTH) or corticosterone can alter behavioural responses to antidepressants, effectively blocking antidepressant efficacy. Herein, we demonstrate that ACTH-(1-24) (100μg/day; 14 days) blocks the immobility-reducing ‘antidepressant’ effects of a single dose of imipramine (10mg/kg) in the forced swim test. This finding was accompanied by altered monoamine tissue levels in the prefrontal cortex (PFC) 1h after exposure to the acute stress of the forced swim test. PFC tissue from ACTH pre-treated animals contained significantly higher serotonin, noradrenaline and adrenaline concentrations relative to saline pre-treated controls. Conversely, dopamine levels were significantly decreased. Altered plasma corticosterone responses to ACTH injections were observed over the treatment course. Measures were taken on treatment days 1, 4, 8, 11, 14 and 15. ACTH administration initially enhanced plasma corticosterone levels, however, these normalised to levels consistent with control animals by day 14. No differences in corticosterone levels were observed across the treatment time course in saline-treated animals. Taken together these results indicate that pre-treatment with ACTH (100μg/day; 14 days) blocks the antidepressant effects of imipramine (10mg/kg), significantly alters key PFC monoamine responses to stress and downregulates glucocorticoid responses. These results suggest that chronic ACTH treatment is a promising paradigm for elucidation of mechanisms mediating antidepressant treatment resistance.