Eugenol as an anti-stress agent: modulation of hypothalamic-pituitary-adrenal axis and brain monoaminergic systems in a rat model of stress

Stress is the leading psychopathological cause for several mental disorders. Physiological and psychological responses to stress are mediated by the hypothalamic?pituitary?adrenal (HPA), sympathoadrenal system (SAS), and brain monoaminergic systems (BMS). Eugenol is reported to substantially modulate brain functions by regulating voltage-gated cation channels and release of neurotransmitters. This study was designed to evaluate the anti-stress effect of eugenol in the 4-h restraint model using rats. Ulcer index was measured as a parameter of the stress response. HPA axis and the SAS were monitored by estimating plasma corticosterone and norepinephrine (NE), respectively. Analysis of NE, serotonin (5-HT), dopamine, and their metabolites in discrete brain regions was performed to understand the role of BMS in the anti-stress effect of eugenol. Stress exposure increased the ulcer index as well as plasma corticosterone and NE levels. Eugenol pretreatment for 7 days decreased the stress-induced increase in ulcer index and plasma corticosterone but not NE levels, indicating a preferential effect on the HPA axis. Furthermore, eugenol showed a ?U?-shaped dose?response curve in decreasing ulcer index and plasma corticosterone levels. Eugenol also reversed the stress-induced changes in 5-HT levels in all brain regions, whereas NE levels were reversed in all brain regions except hippocampus. These results suggest that eugenol possesses significant anti-stress activity in the 4-h restraint model and the effect is due to modulation of HPA and BMS.     

Brain catecholamines and memory modulation: effects of footshock, amygdala implantation, and stimulation

The results of previous studies indicate that the extent of a transient decline in brain norepinephrine (NE) levels shortly after training and administration of any of several memory modulating treatments is correlated with later retention performance. The present experiment assessed such changes after one-trial inhibitory (passive) avoidance training and, in addition, measured concentration changes in 3-methoxy-4-hydroxyphenylglycol (MHPG), the major metabolite of brain NE, as well as dopamine (DA) and epinephrine (EPI) levels. The results indicate that the decreases in brain NE after footshock are accompanied by an increase in MHPG, thus providing additional evidence that brain NE is released after training. DA levels were unchanged after training; brainstem EPI levels increased after the training footshock, but forebrain EPI levels were unchanged. A second experiment examined brain catecholamine levels in animals which received post-training electrical stimulation of the amygdala. The findings of this experiment indicate that the amygdala damage which accompanies electrode implantation apparently results in a chronic change in whole brain NE levels and metabolism. After amygdala, NE concentrations in both brainstem and forebrain samples were reduced by 20% and MHPG was increased by 22-34%. Furthermore, NE levels were not responsive to training in implanted animals. Thus, brain NE levels after training were not predictive of retention performance in amygdala-implanted or -stimulated animals. However, the significance of such findings for understanding the possible role of central NE in memory storage is complicated by the severe modification of the dynamics of brain aminergic systems in animals bearing amygdala electrodes.     

Regional studies of brain biogenic amines in castrated muricidal and non-muricidal wistar rats

Castrated Wistar rats were isolated for 8 months and their muricidal behavior was investigated. Significantly fewer (35%) of such rats became muricidal (CM) compared to controls. The steady-state levels of 5-HT, 5-HIAA, DA, DOPAC, and NE, as well as the changes in synthesis or utilization rats of 5-HT and DA, were analyzed in 15 brain areas derived from CM rats and non-muricidal (CNM) control subjects. In CM rats, higher 5-HT levels were recorded in 5 areas considered to be involved in muricidal behavior: raphe, amygdala, olfactory tubercles, olfactory bulbs, and striatum. The alterations of serotonergic neurotransmission in castrated muricidal rats differ strikingly from those observed in non-castrated muricidal rats. An increase of 5-HT level and in the 5-HT synthesis index as well as a lower 5-HT utilization index were recorded in the raphe of CM rats. Our data suggest that the decrease of 5-HT levels generally said to be the main alteration in the muricidal rat's brain has to be reconsidered. Increased DA levels were observed in CM rats: raphe (50%), amygdala, olfactory tubercles, striatum, and septum (40%), while DA was decreased in cortical areas. There were slight increases of DA synthesis indices in the septum, olfactory tubercles and striatum with a decreased utilization index in the olfactory tubercles. Few alterations in NE levels were observed in CM rats: a decrease in the olfactory tubercles, superior colliculus, and striatum and an increase in temporal cortex. The monoaminergic alterations correlate with the modulation of muricidal behavior. Some areas (the olfactory tubercles, raphe, striatum, and temporal cortex) seem to be particularly involved. © 1992 Wiley-Liss, Inc.     

Regional brain catecholamines and memory: effects of footshock, amygdala implantation, and stimulation

Previous findings have revealed a correlation between post-training release of whole brain norepinephrine (NE) and later retention performance. The present experiment examined changes after a training footshock in NE levels, as well as the levels of the major central NE metabolite, 3-methoxy-4-hydroxyphenylglycol (MHPG), dopamine (DA), and epinephrine (EPI) in eight brain regions. Brain levels of these amines and the metabolite were assessed 10 min after training in a one-trial inhibitory (passive) avoidance task. The results indicate that NE levels decreased significantly in neocortex, neostriatum, hypothalamus, frontal pole, septum, and brainstem, but not in hippocampus or thalamus. The decreases in NE levels were generally accompanied by increases in MHPG; the MHPG/NE ratio increased significantly in all areas in which decreases in NE were observed. DA levels decreased in neostriatum and increased in neocortex and brainstem. Epinephrine levels decreased only in the brainstem sample. Thus, the effects of training on NE are widespread, probably reflecting the release of the amine in most brain regions. Such findings are consistent with the view that posttraining release of brain NE may modulate the storage of new information in many brain regions. One especially potent treatment for modulating memory storage is electrical stimulation of the amygdala. Therefore, we also examined the effects of amygdala implantation and stimulation on brain catecholamine levels to determine whether such changes might be correlated with the effects of amygdala stimulation on memory. The results indicate that electrode implantation into the amygdala results in pervasive changes in NE levels in most brain regions tested. Against this modified baseline, the results of training and electrical stimulation were region specific and very difficult to interpret. The major conclusion which can be derived from this portion of the experiment is that the amygdala damage produced by electrode implantation produces a brain which is substantially different from that of intact animals.     

高唤醒对创伤后应激障碍形成发展的影响及其神经机制

高唤醒是创伤后应激障碍(PTSD)的主要症状之一, 对创伤后应激障碍的形成与发展起核心作用。急性应激期产生的高唤醒可以预测其后PTSD的回避与麻木、再体验等症状的形成, 在创伤后早期, 降低唤醒程度可以减轻PTSD相关的症状表现。下丘脑-垂体-肾上腺轴异常变化会导致去甲肾上腺素(NE)、促肾上腺皮质激素释放因子(CRF)过度释放, 同时皮质醇(酮)水平下降, 这二者是高唤醒产生与维持的主要原因。另外, 5-羟色胺(5-HT)系统的高度激活也影响了高唤醒的形成。食欲素神经肽与NE、CRF与5-HT系统有密切的神经联系, 可能参与高唤醒的调节, 是近年来研究的一个热点。     

Glucocorticoid-induced impairment of long-term memory retrieval in rats: an interaction with dopamine D2 receptors

This study investigated glucocorticoid-dopaminergic interactions in modulating retrieval of long-term memory in an inhibitory avoidance task. Young adult male rats were trained in one trial inhibitory avoidance task (0.5 mA, 3 s footshock). On the retention test given 48 h after training, the latency to re-enter the dark compartment of the apparatus was recorded. Systemically administered corticosterone (1 or 3 mg/kg) given to rats 30 min before retention testing impaired their memory retrieval, but the lower dose was more effective than the higher one. Administration of the dopamine (DA) D2 receptor antagonist sulpiride (6 or 20 mg/kg) 30 min before corticosterone attenuated the impairing effects of corticosterone (1 mg/kg) on memory retrieval. Administration of the DA D1 receptor antagonist SCH23390 (25 or 50 microg/kg) had no effect on corticosterone-induced impairment of memory retrieval. Further, applied doses of sulpiride or SCH23390 alone were ineffective in modulating memory retrieval. These findings provide evidence for the existence of an interaction between glucocorticoids and DA D2 receptor on memory retrieval process.     

Effects of repetitive hypoglycemia on neuroendocrine response and brain tyrosine hydroxylase activity in the rat

Hypoglycemia-associated autonomic failure (HAAF) is a syndrome of acute adaptation to a metabolic stressor, in which neuroendocrine responses to repetitive hypoglycemic bouts are blunted. The CNS mechanisms that contribute to HAAF are unknown. In the present study, we modeled HAAF in the rat and measured the activity of tyrosine hydroxylase (TH) as an index of acute noradrenergic activation, to test the hypothesis that noradrenergic activation of the hypothalamus might be impaired. In association with a significant counter-regulatory response to a single bout of hypoglycemia (elevated corticosterone, catecholamines, and glucagon), TH activity was elevated overall in brainstem NE cell body areas and hypothalamus. With multiple hypoglycemic episodes in a 24 h period, the counter-regulatory response was blunted, and hypothalamic TH activity was comparable to that of saline-infused controls. In a similar paradigm, multiple bouts of CNS neuroglucopenia did not blunt the hyperglycemic or corticosterone responses, and were required for elevation of TH activity. This alternate response pattern suggests that insulin-induced hypoglycemia and cerebral neuroglucopenia represent somewhat different metabolic stressors at the CNS.     

Catecholamine depletion in mice upon reexposure to stress: mediation of the escape deficits produced by inescapable shock

Immediately following exposure to 60 inescapable shocks, Swiss-Webster mice had significantly reduced hypothalamic norepinephrine (NE). Within 24 hr NE levels returned to control values. Reexposure to as few as 10 shocks 24 hr after initial stress exposure resulted in a significant decline of hypothalamic NE. Moreover, at this interval after inescapable shock, escape performance was severely disrupted, with a large proportion of mice exhibiting numerous failures to escape shock. Increasing brain dopamine (DA) and NE by L-dopa treatment prior to inescapable shock prevented the escape deficits. Conversely, pairing five inescapable shocks with NE depletion by FLA-63, or both DA and NE depletion by alpha-methyl-p-tyrosine, disrupted escape performance 24 hr later. Residual drug effects, state dependence, or sustained amine turnover could not account for the behavioral changes observed. Data are discussed in terms of catecholamine mediation of escape performance through variations in response maintenance abilities. Furthermore, it is suggested that the long-term effects of inescapable shock may be due to sensitization effects or conditioned amine depletion.     

Post-training brain catecholamine levels: lack of response to water-motivated training

Rats were trained in a one-trial appetitive task using water motivation. Brain catecholamine and metabolite levels were assessed in samples collected 10 min after training. There was no evidence that brain NE levels were modified by training, although catecholamine levels increased when the animals were placed in a novel environment. These results differ from those obtained after avoidance training where the extent of a post-training decrease in brain norepinephrine predicts later retention performance.     

Requirement of dopamine signaling in the amygdala and striatum for learning and maintenance of a conditioned avoidance response

Two-way active avoidance (2WAA) involves learning Pavlovian (association of a sound cue with a foot shock) and instrumental (shock avoidance) contingencies. To identify regions where dopamine (DA) is involved in mediating 2WAA, we restored DA signaling in specific brain areas of dopamine-deficient (DD) mice by local reactivation of conditionally inactivated Th genes using viral gene therapy. Among all targeted areas--prefrontal cortex (PFC), amygdala, ventral striatum, dorsal striatum, and whole striatum--only restoration of DA signaling to both the whole striatum together with the amygdala enabled DD mice to acquire 2WAA. However, after prolonged overtraining during which DD mice had DA synthesis systemically reconstituted pharmacologically with L-3,4-dihydroxyphenylalanine (L-Dopa), DA signaling in the striatum alone was sufficient to maintain 2WAA, whereas DA signaling in the PFC together with the amygdala was insufficient to maintain 2WAA. Our results indicate that learning 2WAA requires DA signaling in both the amygdala and the entire striatum; however, after sufficient training, DA signaling in the striatum alone can maintain the learned avoidance behavior.     

Microinjections of the dopamine D2 receptor antagonist sulpiride into the medial prefrontal cortex attenuate glucocorticoid-induced impairment of long-term memory retrieval in rats

We recently reported that blockade of dopamine (DA) D2 receptors attenuated deficits in long-term memory retrieval induced by a systemic injection of corticosterone, but the anatomical sites of such interaction were not known. In this study, we investigated whether the DA D2 receptors located in the medial prefrontal cortex (mPFC) may play a role in the impairing effects of glucocorticoids on the memory retrieval process. Young adult male rats were trained in a one trial inhibitory avoidance task (0.5 mA, 3s footshock). On the retention test given 48 h after training, the latency to re-enter the dark compartment and the time spent in light compartment of the apparatus were recorded. Systemically administered corticosterone (1mg/kg) given to rats 30 min before retention testing impaired their memory retrieval. Bilateral microinjections of the DA D2 receptor antagonist sulpiride (10 or 100 ng/0.5 microl per side) into the mPFC 30 min before corticosterone administration attenuated the glucocorticoid-induced impairment of memory retrieval. Furthermore, applied doses of sulpiride alone were ineffective in modulating memory retrieval. These findings indicate that D2 receptors located in the mPFC play an important role in mediating the impairing effects of glucocorticoids on memory retrieval.     

Exploration and avoidance learning after ibotenic acid and radio frequency lesions in the rat amygdala

Open-field activity, avoidance behavior, and plasma corticosterone levels were studied after intraamygdala injections of 3.0 micrograms ibotenic acid (IBO) and radio-frequency (RF) lesions in the amygdala complex of male Wistar rats. The experiments were undertaken to evaluate the importance of amygdala neurons versus axons of passage in fear-motivated behavior. The IBO lesions led to increased open-field activity, but no impairments in active avoidance learning, nor changes in basal or experimental levels of plasma corticosterone. The RF lesions, on the other hand, led to an increase in experimental plasma corticosterone levels. In the one-way avoidance task the RF lesions, in contrast to the IBO lesions, led to significant impairments in the acquisition of the avoidance response. Although the long-term axon-sparing effect of IBO is questioned since cavities were detected in the affected areas 8 weeks after the injections, the differences in avoidance learning and in corticosterone levels between the RF and the IBO lesions indicate that the axons were functionally active at the time of testing (14-26 days postoperatively). The increase in open-field activity is attributed to the destruction of amygdala neurons and neurons in the overlying cortex, while an avoidance deficit seem to depend on the destruction of axons. On the basis of the behavioral results and the corticosterone data in these experiments, it is suggested that the behavioral changes are not attributable to a general reduction in the arousal of fear. However, since the IBO lesions did not affect the most medial parts of the amygdala complex including the central amygdala nucleus, the role of this nucleus in fear arousal has to be investigated further.     

Chronic sweet intake lowers pain thresholds without changing brain mu- or delta-opiate receptors

Sprague-Dawley rats drank sweetened (3% dextrose + 0.144% saccharin, w/v) or unflavored water for 18 days and subsequent pain reactivity was assessed using a hot plate. Compared to the rats that consumed unflavored water, the rats that consumed sweet water responded more quickly on the hot plate indicating that their threshold for pain was lowered. Another group of rats given identical exposure to the fluids had their brains prepared for measuring opiate receptor binding using the delta-receptor ligand [3H]D-Ala-D-Leu-enkephalin ([3H]DADLE) and the mu-receptor selective ligand [3H]Tyr-D-Ala-Gly-MePhe-Gly-ol ([ 3H]DAGO). Binding of these opiates to mu- and delta-receptors in the cerebral cortex, striatum, hippocampus, hypothalamus, brain stem, and remaining brain regions was the same for the rats that drank sweet fluids and those that drank unflavored water. These findings suggest that drinking sweet fluids lowers pain thresholds but does not alter mu- and delta-receptors.     

Glucocorticoid administration into the dorsal striatum [corrected] facilitates memory consolidation of inhibitory avoidance training but not of the context or footshock components

It is well established that glucocorticoid administration into a variety of brain regions facilitates memory consolidation of fear-conditioning tasks, including inhibitory avoidance. The present findings indicate that the natural glucocorticoid corticosterone administered into the dorsal striatum (i.e., caudate nucleus) of male Wistar rats produced dose- and time-dependent enhancement of inhibitory avoidance memory consolidation. However, as assessed with a modified inhibitory avoidance procedure that took place on two sequential days to separate context training from footshock training, corticosterone administration into the dorsal striatum did not enhance memory of either the contextual or aversively motivational aspects of the task.     

Central catecholamine and peripheral noradrenaline depletion by 6-hydroxydopamine and active avoidance learning in rats

Three experiments are reported on the relation among catecholamine (CA) depletion, plasma corticosterone (11-OHCS) levels, and active aversive learning. A baseline study showed no significant relation beteen resting levels of 11-OHCS and central and/or peripheral CA depletion. On a two-way avoidance learning task, depletion of brain CAs produced a widespread learning deficit. When both central and peripheral CAs were depleted, only escape learning emerged. Similar effects occurred on a one-way task, but differences were not so marked. The findings are discussed in terms of a theory of CA-dependent cues.     

Transient overexpression of the 5-HT1A receptor impairs water-maze but not hole-board performance

Previously, we showed that mice that overexpress the 5-HT(1A) receptor transiently from embryonic to perinatal stages show reduced anxiety and changes in brain serotonin turnover as adults. Here, we investigated the long-term effects of the temporary overexpression of the 5-HT(1A) receptor during early embryonic and perinatal development on the performance in two memory tasks. In the hole-board test mice that were homozygous for the transgene showed similar behavioral habituation but increased locomotion compared to heterozygous mice. In contrast water-maze performance of homozygous mice was impaired compared to heterozygous mice. These results suggest that a transient overexpression of 5-HT(1A) receptor during embryonic and perinatal development has detrimental effects on water-maze performance at adult stages.     

Septal lesions and behavior: effects of presurgical rearing and strain of mouse.

Male mice (C57BL/10J or SJL/J strains) were reared in either enriched social cages or restricted individual cages from 25 days of age until they underwent septal or control surgery 1 mo later. Enrichment differentially altered septal or control behavior as measured by: fluid consumption of water, saccharin, and quinine; performance on a rotorod; and the acquisition of an active avoidance task. The interactions of presurgical history with brain damage were manifested differently in the two strains of mice. The importance of attending more to genetic and presurgical history in attempts to define the effects of brain damage on behavior and to determine the function of brain structures is discussed.     

Strain-dependent interactions between MK-801 and cocaine on retention of C57BL/6 and DBA/2 mice tested in a one-trial inhibitory avoidance task: involvement of dopaminergic mechanisms

Two sets of experiments were carried out with C57BL/6 (C57) and DBA/2 (DBA) mice tested in a one-trial inhibitory avoidance task. In the first set C57 and DBA mice were injected posttraining with saline or with the D1 DA receptor antagonist SCH 23390 and then with saline, cocaine (5 mg/kg), MK-801 (0.1 mg/kg), or with a combination of these two drugs. Cocaine enhanced retention in the C57 strain and impaired it in the DBA strain, and MK-801 potentiated the effects of cocaine in both strains. Furthermore, pretreatment with SCH 23390 completely antagonized the potentiation of the effects of cocaine exerted by MK-801. In the second set of experiments mice belonging to these same two strains were injected posttraining with vehicle or with the D2 DA receptor antagonist (-)-sulpiride and then with saline, cocaine (5 mg/kg), MK-801 (0.1 mg/kg), or with a combination of these two drugs. Pretreatment with the D2 DA receptor antagonist completely antagonized in both strains the potentiation of the effect of cocaine exerted by MK-801. The results of the present research show that the noncompetitive NMDA receptor antagonist MK-801 enhances the effect of cocaine on retention performance in C57 and DBA mice and that dopaminergic mechanisms are involved in this potentiation.     

Exposure to a retrieval cue in rats induces changes in regional brain glucose metabolism in the amygdala and other related brain structures

Pre-test exposure to training-related cues is known to improve subsequent retention performance. To identify brain regions engaged in processes promoted by retrieval cues, a brain imaging approach using the [6-14C]glucose autoradiographic technique was used. Sprague-Dawley rats trained in a brightness discrimination avoidance task were submitted to different cueing conditions after a 1- or a 21-day training-to-test interval (TTI). Animals were either non-cued, cued with a box, or cued with a box and the light that served as a discriminative stimulus. Effects of the different cueing conditions on retention performance or on metabolic activity in 58 different brain regions were investigated. Rats cued with the light exhibited a subsequent improvement of their retention performance relative to controls, when tested at the 1- but not 21-days TTI, confirming our previous results. At the 1-day retention interval, a comparison between rats cued with the box and rats cued with the box and the light showed that the light cue significantly increased glucose uptake in a neuronal network composed of the lateral, basal, and central nuclei of the amygdala, the anterior and suprachiasmatic hypothalamic nuclei, the nucleus accumbens, the medial septum, and the insular cortex. In contrast, at the 21-day retention interval, both groups demonstrated similar cerebral metabolic activity. The present results indicate that exposure to a light cue increased metabolic activity in the previously mentioned brain structures only when the light acted as an effective retrieval cue, suggesting an involvement of this network in the processes triggered by a retrieval cue. Arguments are provided supporting the notion that the amygdala may play a key role in these processes. Whether the amygdala is a part of a neural network involved in retrieval processes or in neuromodulating systems that favour the efficacy of retrieval processes is also discussed.     

The neural correlates of driving performance identified using positron emission tomography

Driving is a complex behavior involving multiple cognitive domains. To identify neural correlates of driving performance, [15O]H2O positron emission tomography was performed using a simulated driving task. Compared with the resting condition, simulated driving increased regional cerebral blood flow (rCBF) in the cerebellum, occipital, and parietal cortices. Correlations between rCBF and measurements of driving performance were evaluated during simulated driving. Interestingly, rCBF in the thalamus, midbrain, and cerebellum were positively correlated with time required to complete the course and rCBF in the posterior cingulate gyrus was positively correlated with number of crashes during the task. These brain regions may thus play roles in the maintenance of driving performance.