Strain differences in the effects of chronic corticosterone exposure in the hippocampus.

Stress hormones are thought to be involved in the etiology of depression, in part, because animal models show they cause morphological damage to the brain, an effect that can be reversed by chronic antidepressant treatment. The current study examined two mouse strains selected for naturalistic variation of tissue regeneration after injury for resistance to the effects of chronic corticosterone (CORT) exposure on cell proliferation and neurotrophin mobilization. The wound healer MRL/MpJ and control C57BL/6J mice were implanted subcutaneously with pellets that released CORT for 7 days. MRL/MpJ mice were resistant to reductions of hippocampal cell proliferation by chronic exposure to CORT when compared to vulnerable C57BL/6J mice. Chronic CORT exposure also reduced protein levels of brain-derived neurotrophic factor (BDNF) in the hippocampus of C57BL/6J but not MRL/MpJ mice. CORT pellet exposure increased circulating levels of CORT in the plasma of both strains in a dose-dependent manner although MRL/MpJ mice may have larger changes from baseline. The strains did not differ in circulating levels of corticosterone binding globulin (CBG). There were also no strain differences in CORT levels in the hippocampus, nor did CORT exposure alter glucocorticoid receptor or mineralocorticoid receptor expression in a strain-dependent manner. Strain differences were found in the N-methyl-D-aspartate (NMDA) receptor, and BDNF I and IV promoters. Strain and CORT exposure interacted to alter tropomyosine-receptor-kinase B (TrkB) expression and this may be a potential mechanism protecting MRL/MpJ mice. In addition, differences in the inflammatory response of matrix metalloproteinases (MMPs) may also contribute to these strain differences in resistance to the deleterious effects of CORT to the brain.

Strain-dependent effects of diazepam and the 5-HT2B/2C receptor antagonist SB 206553 in spontaneously hypertensive and Lewis rats tested in the elevated plus-maze

The 5-HT2B/2C receptor antagonist SB 206553 exerts anxiolytic effects in rat models of anxiety. However, these effects have been reported for standard rat strains, thus raising the issue of SB 206553 effects in rat strains displaying different levels of anxiety. Herein, the effects of SB 206553 in a 5-min elevated plus-maze test of anxiety were compared to those of the reference anxiolytic, diazepam, in two rat strains respectively displaying high (Lewis rats) and low (spontaneously hypertensive rats, SHR) anxiety. Diazepam (0.37, 0.75, or 1.5 mg/kg; 30 min before testing) increased in a dose-dependent manner the behavioral measures in SHR, but not in Lewis rats. On the other hand, SB 206553 (1.25, 2.5, or 5 mg/kg; 30 min before testing) failed to alter the anxiety parameters in both strains, whereas it increased closed arm entries in Lewis rats, suggesting that it elicited hyperactivity in the latter strain. Accordingly, the hypolocomotor effect of the nonselective 5-HT2B/2C receptor agonist m-chlorophenylpiperazine (1.5 mg/kg ip 20 min before a 15-min exposure to an activity cage) was prevented by the 1.25 and 2.5 mg/kg doses of SB 206553 in Lewis rats and SHR, respectively. Compared with SHR, Lewis rats may display a lower response to benzodiazepine-mediated effects and a more efficient control of locomotor activity by 5-HT2B/2C receptors

Strain-dependent effects of diazepam and the 5-HT2B/2C receptor antagonist SB 206553 in spontaneously hypertensive and Lewis rats tested in the elevated plus-maze.

The 5-HT2B/2C receptor antagonist SB 206553 exerts anxiolytic effects in rat models of anxiety. However, these effects have been reported for standard rat strains, thus raising the issue of SB 206553 effects in rat strains displaying different levels of anxiety. Herein, the effects of SB 206553 in a 5-min elevated plus-maze test of anxiety were compared to those of the reference anxiolytic, diazepam, in two rat strains respectively displaying high (Lewis rats) and low (spontaneously hypertensive rats, SHR) anxiety. Diazepam (0.37, 0.75, or 1.5 mg/kg; 30 min before testing) increased in a dose-dependent manner the behavioral measures in SHR, but not in Lewis rats. On the other hand, SB 206553 (1.25, 2.5, or 5 mg/kg; 30 min before testing) failed to alter the anxiety parameters in both strains, whereas it increased closed arm entries in Lewis rats, suggesting that it elicited hyperactivity in the latter strain. Accordingly, the hypolocomotor effect of the nonselective 5-HT2B/2C receptor agonist m-chlorophenylpiperazine (1.5 mg/kg ip 20 min before a 15-min exposure to an activity cage) was prevented by the 1.25 and 2.5 mg/kg doses of SB 206553 in Lewis rats and SHR, respectively. Compared with SHR, Lewis rats may display a lower response to benzodiazepine-mediated effects and a more efficient control of locomotor activity by 5-HT2B/2C receptors.

In vivo actions of the selective 5-HT1A receptor agonist BAY x 3702 on serotonergic cell firing and release.

We investigated the effects of the novel 5-HT1A receptor agonist BAY x 3702 on the serotonergic function in rat brain using single unit recordings in the dorsal raphe nucleus (DR) of anesthetized rats and in vivo microdialysis in freely moving rats. The administration of BAY x 3702 (0.25-4 microg/kg i.v.) suppressed the firing activity of 5-HT neurones. This effect was antagonized by a low dose of the selective 5-HT1A receptor antagonist WAY 100635 (5 microg/kg i.v.). In microdialysis experiments, BAY x 3702 (10-100 microg/ kg s.c.) reduced dose-dependently the 5-HT output in the dorsal and median raphe (MnR) nucleus, dorsal hippocampus (DHPC) and medial prefrontal cortex (mPFC) in a regionally selective manner. Maximal effects were observed in the MnR and mPFC, with reductions to approximately 15% of baseline at a dose of 0.1 mg/kg s.c. The decrease in 5-HT output produced in the DR and DHPC was more moderate, to 45% of baseline at 0.1 mg/kg s.c. BAY x 3702. WAY 100635 (0.3 mg/kg s.c.) completely antagonized the effect of BAY x 3702 (30 microg/kg s.c.). The application of BAY x 3702 in the DR (1-100 microM) reduced the local 5-HT output to 25% of baseline. In rats implanted with two dialysis probes (in DR and mPFC) the application of BAY x 3702 (30 microM) in the DR reduced the 5-HT output in the DR and that in mPFC. These effects were significantly antagonized by the co-perfusion of WAY 100635 (100 microM) in the DR. Overall, these results indicate that the systemic administration of BAY x 3702 reduces the 5-HT release with high potency through the activation of midbrain 5-HT1A receptors.

Effects of repeated fluoxetine on anxiety-related behaviours, central serotonergic systems, and the corticotropic axis axis in SHR and WKY rats.

In keeping with the anxiolytic property of selective serotonin reuptake inhibitors (SSRIs) in humans, we have examined in the spontaneously hypertensive rat (SHR) and the Wistar-Kyoto (WKY) rat, which display low and high anxiety, respectively, some psychoneuroendocrine effects of a repeated treatment with the SSRI fluoxetine (5 or 10 mg/kg daily, for 3 weeks). Two days after the last injection, plasma levels of fluoxetine were not detectable whereas those of its metabolite, norfluoxetine, were present to similar extents in both strains. By means of the elevated plus-maze test (29-30 h after the 13th administration of fluoxetine) and an open field test (48 h after the last injection of fluoxetine), it was observed that fluoxetine pretreatment did not yield anxiolysis; hence, some, but not all, behaviours were indicative of anxiety and hypolocomotion (as assessed through principal component analyses and acute diazepam studies). In both strains, the 10 mg/kg dose of fluoxetine decreased hypothalamus 5-HT and 5-HIAA levels, and reduced midbrain and/or hippocampus [3H]citalopram binding at 5-HT transporters, but did not affect [3H]8-hydroxy-2-(di-N-propylamino)tetralin binding at hippocampal 5-HT1A receptors. However, the fluoxetine-elicited reduction in hippocampal 5-HT transporter binding was much more important in WKY than in SHR rats, this strain-dependent effect being associated in WKY rats with a reduction in cortical [3H]ketanserin binding at 5-HT2A receptors. Lastly, in WKY rats, repeated fluoxetine administration increased adrenal weights and the plasma corticosterone response to open field exposure, but did not affect the binding capacities of hippocampal mineralocorticoid and glucocorticoid receptors. These data show that key psychoneuroendocrine responses to repeated fluoxetine administration may be strain-dependent, and that repeated fluoxetine administration does not yield anxiolysis, as assessed by two standard tests of emotivity.

Serotonin and stress.

Forty-five years after its discovery, brain serotonin (5-HT) is still the subject of intense research aimed at understanding its role in stress adaptation. At the presynaptic level, numerous stressors increase nerve firing and extracellular 5-HT at the level of serotonergic cell bodies or nerve terminals. Different studies have reported stressor- and region-specific changes in extracellular 5-HT, a view challenged by electrophysiological and neurochemical evidence for a nonspecific response of serotonergic neurones to stressors when activity/arousal is taken into account. In addition, early studies indicate that stress-induced elevation in 5-HT synthesis, a key counter-regulatory process allowing serotonergic homeostasis, is mediated by specific neuroendocrine mechanisms. In addition to the multiplicity of postsynaptic 5-HT receptors and their specific regulation by corticoids, specificity to stressors is also underscored when considering one receptor type such as the 5-HT1A receptor. Stress studies should consider the past experience and the genetic status of the individual as key modulators of the serotonergic responses to stress.

Behavioral, neuroendocrine and serotonergic consequences of single social defeat and repeated fluoxetine pretreatment in the Lewis rat strain.

We have analysed some behavioral, neuroendocrine and serotonergic consequences of a single (30-min) social defeat followed by 14-18 h of sensory contact with the aggressor, in Lewis rats, an inbred strain highly sensitive to chronic social stressors [Berton O. et al. (1998) Neuroscience 82, 147-159]. In addition, we have investigated how the aforementioned consequences are affected by pretreatment with the selective serotonin reuptake inhibitor, fluoxetine (7.5 mg/kg/day for 21 days). A single social defeat triggered hypophagia and body weight loss, and increased anxiety in the elevated plus-maze. It did not affect baseline plasma adrenocorticotropic hormone levels and renin activity, but decreased plasma corticosterone levels. On the other hand, the responses of the latter variables to subsequent acute forced swim stress were blunted (corticosterone) or amplified (adrenocorticotropic hormone, renin activity) by prior defeat. The density of hippocampal serotonin transporters, but not that of hippocampal serotonin-1A and cortical serotonin-2A receptors, was decreased by a single social defeat; in addition, neither tryptophan availability and serotonin synthesis/metabolism, nor serotonin-1A autoreceptor-mediated functions (inhibition of serotonin synthesis, hyperphagia) were affected. Fluoxetine pretreatment diminished social defeat-induced hypophagia, body weight loss and anxiety without affecting these variables in control animals. This pretreatment increased plasma corticosterone levels in resting and acutely stressed rats, but abolished social defeat-elicited corticosterone hyporesponsiveness to acute forced swim stress. Except for a decrease in midbrain serotonin transporter density, fluoxetine did not affect the other serotonergic indices analysed herein, i.e. serotonin-1A and serotonin-2A receptor densities, serotonin synthesis/metabolism. A single social defeat in Lewis rats produces behavioral and endocrine alterations that may model some aspects of human anxiety disorders. In this paradigm, prior fluoxetine treatment is endowed with adaptive behavioral, and possibly neuroendocrine, effects without affecting the key elements of central serotonergic systems analysed herein.

Differential effects of social stress on central serotonergic activity and emotional reactivity in Lewis and spontaneously hypertensive rats.

Social stress by repeated defeat has been shown to be endowed with neuroendocrine and behavioural effects that render this stress model useful to identify adaptive mechanisms. Among these mechanisms, those related to central serotonergic systems (e.g., hippocampal 5-HT1A receptors, cortical 5-HT2A receptors) have been particularly underlined. Nonetheless, how (i) the neuroendocrine and behavioural effects of social stress are affected by the genetic status of the animal, and (ii) this status affects the relationships between central serotonergic systems and adaptive processes has not been studied so far. The present study has thus analysed the effects of repeated defeat (once a day for seven days) by Long-Evans resident rats upon the psychoneuroendocrine profile of Lewis rats and spontaneously hypertensive rats previously characterized for their contrasting social and anxiety-related behaviours. Repeated defeat decreased in a time-dependent manner, body weight growth and food intake in both strains, these decreases being, however, more severe and longer lasting in Lewis rats. This strain-dependent difference could not be accounted for by differences in physical contacts with the resident rats as the number of attacks and their latency throughout the stress period were similar between spontaneously hypertensive and Lewis rats. When exposed to an elevated plus-maze test of anxiety, the unstressed Lewis rats entered less the open arms than their spontaneously hypertensive counterparts, thus confirming that Lewis rats are more anxious than spontaneously hypertensive rats. This difference was amplified by stress as the latter increased anxiety-related behaviours in Lewis rats only. These strain- and stress-related differences were associated with differences in locomotor activity, this being increased in unstressed Lewis compared with spontaneously hypertensive rats; moreover, stress triggered hypolocomotion in the former but not the latter strain. Lastly, in the forced swimming test. Lewis rats spent more time immobile than spontaneously hypertensive rats with stress increasing immobility in a strain-independent manner. Beside the aforementioned metabolic changes, the activity of the hypothalamo-pituitary-adrenal axis was slightly stimulated in a strain-independent manner by the stressor, as assessed by increased corticosterone levels and adrenal weights, and decreased thymus weights. In Lewis, but not in spontaneously hypertensive rats, midbrain serotonin metabolism was increased by stress, a difference associated with an increased Bmax value of cortical [3H]ketanserin binding at 5-HT2A receptors. On the other hand, the Bmax value of hippocampal [3H]8-hydroxy-2-(di-n-propylamino)tetralin binding at 5-HT1A receptors was decreased by stress, this reduction being amplified in spontaneously hypertensive compared with Lewis rats. This study shows that the psychoneuroendocrine responses to social stress may have a genetic origin, and that the use of socially stressed Lewis and spontaneously hypertensive rats may provide an important paradigm to study adaptive processes. However, whether the aforementioned strain-dependent differences in central serotonergic systems (partly or totally) underlie the distinct profiles of emotivity measured in spontaneously hypertensive and Lewis rats, is discussed in the context of the relationships between serotonergic systems and behavioural responses to novel environments.

Central serotonergic systems in the spontaneously hypertensive and Lewis rat strains that differ in the elevated plus-maze test of anxiety.

The spontaneously hypertensive (SHR) and Lewis (LEW) strains differ in numerous behavioral tests, including the elevated plus-maze. In keeping with the crucial role of central serotonin (5-HT) in anxiety, we checked for strain differences regarding several determinants of 5-HT activity. In addition to confirming that LEW rats displayed anxious behaviors in the plus-maze compared with SHR, we found that in vitro, central tryptophan hydroxylase activity was higher in LEW rats than in SHR. However, ex vivo studies in midbrains and hippocampi revealed that neither 5-HT synthesis nor 5-HT and 5-hydroxyindoleacetic acid levels differed between strains. [3H]8-Hydroxy-2-(di-n-pro-pylamino)tetralin binding at midbrain 5-HT1A autoreceptors and hippocampal 5-HT1A postsynaptic receptors, [3H]ketanserin binding at cortical and striatal 5-HT2A receptors and [3H]citalopram binding at midbrain and hippocampal 5-HT transporters did not vary between strains. The inhibition of 5-HT synthesis by 5-HT1A autoreceptor stimulation was similar in both strains. Forepaw treading and flat body posture after 5-HT1A postsynaptic receptor stimulation were higher and lower, respectively, in SHR than in LEW rats. Last, 1-(4-iodo-2,5-dimethoxy-phenyl)-2-aminopropane- and quipazine-elicited head shakes, a 5-HT2A receptor-mediated response, were increased in the SHR strain compared with the LEW strain; on the other hand, 1-(3-chlorophenyl)piperazine triggered similar 5-HT2B/2C receptor-mediated decreases in motor activity in the two strains. This study shows that although the low-anxiety (SHR) and high-anxiety (LEW) strains vary in some aspects of 5-HT function, key components such as the 5-HT1A autoreceptors are not different.

Effects of food deprivation on midbrain 5-HT1A autoreceptors in Lewis and SHR rats.

Food deprivation stimulates the activity of the hypothalamo-pituitary-adrenal axis and brain serotonin (5-hydroxytryptamine, 5-HT) synthesis. Because midbrain somato-dendritic 5-HT1A autoreceptors may obey homologous and heterologous (e.g. by glucocorticoids) down-regulation, we have analyzed whether 24 hr of fasting affects midbrain 5-HT1A receptor binding and sensitivity in Lewis and SHR rats (i.e. strains that differ in behavioral/neuroendocrine responses to stressors). Fasting affected neither [3H]8-hydroxy-2-(di-N-propylamino)tetralin ([3H]8-OH-DPAT) binding at 5-HT1A autoreceptors nor 8-OH-DPAT-induced inhibition of midbrain 5-HT synthesis (an index of 5-HT1A autoreceptor sensitivity). Because fasting increased 5-HT precursor (tryptophan) levels to similar extents in the midbrains of saline- and 8-OH-DPAT-treated rats, we conclude that food deprivation does not affect 5-HT1A autoreceptors. In turn, our results suggest that the differential effects of 5-HT1A receptor agonists on food intake, in fed and fasted rats may be independent from 5-HT1A autoreceptors.

A multiple-test study of anxiety-related behaviours in six inbred rat strains.

Recent studies have underlined the impact of genetic factors in anxiety profiles. In this context, we have initiated a series of experiments aiming to select, among six inbred strains of rats, a pair of strains that contrasts the most in fear-related behaviours measured in the open field, the elevated plus-maze, the black and white box and the social interaction test. Significant interstrain differences were found for all behavioural measures. A factor analysis of all variables produced three independent factors explaining 85.1% of the total variance. Factor 1 had high loadings from variables related to the approach/avoidance towards aversive stimuli (e.g., center of the open field, open arms of the plus-maze and white compartment of the black/white box). Variables related to general activity in novel environments (e.g., total locomotion in the open field and closed-arm entries in the plus-maze) loaded highly on Factor 2. Defecation and time of social interaction loaded positively on Factor 3. To verify whether elevated plus-maze variables loading on Factor 1 were associated to anxiety, the effects of single doses of diazepam and pentylenetetrazole were examined in Lewis and SHR rats, i.e., the most contrasting strains regarding Factor 1. Variables with high loadings on this factor changed in opposite ways in response to diazepam and pentylenetetrazole treatments. This study suggests, thus, that Lewis and SHR strains may constitute a useful model for studying the neurobiological mechanisms underlying the interindividual differences in baseline levels of anxiety.

Behavioral reactivity to social and nonsocial stimulations: a multivariate analysis of six inbred rat strains.

Male rats from six inbred rat strains (Spontaneously Hypertensive Rat, Wistar Kyoto, Brown Norway, Wistar Furth, Fischer 344, and Lewis) have been compared for their behavioral reactivity when placed in several nonsocial (elevated plus-maze, open field) and social (social interaction in aversive and neutral environment, resident-intruder test, chronic social stress) settings. In addition, a factorial analysis was performed to assess how the variables measured in these different tests related to each other. Besides significant strain-related differences in all tests, the factorial analysis showed that, in nonsocial environments, the strains contrasted essentially along two independent behavioral traits, the propensity to approach or avoid an aversive stimulus and general motor activity in novel environments (two indices of emotionality). In the social settings, marked interstrain differences were observed regarding the expression of aggressive behaviors but these differences were not related to the respective levels on the two nonsocial components of reactivity. Furthermore, large genetic differences were observed in variations of body weight induced by a chronic social stressor paradigm. The factorial analysis suggested a lack of relationship between the effect of social stressors on body weight and the measures of emotionality and general activity obtained in the nonsocial tests. Conversely, these variations were influenced by the levels of aggressiveness and sociability. Taken together, these results show (i) that the behavioral variability observed in rats in social and nonsocial environments, is influenced by genetic factors and (ii) that the behavioral reactivity to social stimulations is a specific feature, dissociable from the levels of the different components of emotionality (approach/avoidance and general activity) as evaluated by the behavioral responses to nonsocial settings.