Dissociation between mesocortical dopamine release and fear-related behaviours in two psychogenetically selected lines of rats that differ in coping strategies to aversive conditions

The mesocortical and mesolimbic dopaminergic (DAergic) pathways are activated by either aversive or rewarding stimuli. The functional tone of these DAergic neurons also increases during the execution of cognitive tasks. The present study was designed to examine the relationship between mesocortical and mesolimbic DAergic function and the expression of fear-related behaviours as compared with attention- and cognition-related mechanisms (e.g. coping strategies), in response to aversive conditions. To this aim, we used two psychogenetically selected rat lines, Roman high-avoidance (RHA/Verh) and Roman low-avoidance (RLA/Verh), which display drastically different emotion- and coping-related behaviours in response to stressors: RLA/Verh rats are 'reactive copers' and more fearful than RHA/Verh rats, which are 'proactive copers'. Brain dialysis experiments demonstrated that tail-pinch (TP) and the anxiogenic compounds pentylenetetrazol (PTZ) and ZK 93426 increased DA output in the medial prefrontal cortex (PFCX) of RHA/Verh but not RLA/Verh, rats. In contrast, in the shell compartment of the nucleus accumbens (NAC shell), TP caused a small increase in DA output only in RLA/Verh rats, whereas PTZ and ZK 93426 had no significant effect on either line. RHA/Verh rats displayed more robust and longer lasting coping activity and less frequent freezing and self-grooming episodes than did RLA/Verh rats after TP, PTZ or ZK 93426. This dissociation between fear-related behaviour and cortical DAergic activation argues against the view that the latter may be involved in the control of fear-like responses. We therefore propose that the activation of mesocortical DAergic projections by aversive stimuli underlies the cognitive mechanisms that are triggered in an attempt to gain control over the stressor.     

Differential effects of embryo transfer and maternal factors on anxiety-related behavior and numbers of neuropeptide Y (NPY) and parvalbumin (PARV) containing neurons in the amygdala of inbred C3H/HeN and DBA/2J mice

The effects of reciprocal embryo transfers were studied on anxiety-related behavior of inbred C3H/HeN and DBA/2J mice on the elevated plus maze (EPM), and related to amygdaloid neuropeptide Y (NPY)- and parvalbumin (PARV)-immunoreactive neurons. Embryo transfer significantly reduced closed arm entries in in-stain-transferred C3H mice, and maternal factors influenced open arm entries only in interaction with genetic background and sex. In DBA/2J-mice, embryo transfer resulted in a reduced number of NPY-immunoreactive (NPY-ir) neurons, while PARV-immunoreactive (PARV-ir) cells were not affected. In C3H/HeN mice, however, in-strain embryo transfer only resulted in a reduction of the number of PARV-immunoreactive neurons. Maternal factors mainly induced changes in the number of NPY-ir neurons in the basolateral complex of the amygdala either directly or in interaction with genetic factors. In summary, in-strain embryo transfer had a minor effect on the behavior of C3H/HeN mice, and a differential influence on the numbers of amygdaloid NPY-ir and PARV-ir neurons of inbred C3H/HeN and DBA/2J mice. Maternal factors had a stronger impact on the numbers of NPY-ir neurons than PARV-ir neurons. The present results indicate that alterations in behavior and amygdala morphology induced by embryo transfer or maternal factors depend on the genetic background of the mouse strains used.     

GABAergic and dopaminergic transmission in the brain of Roman high-avoidance and Roman low-avoidance rats

The GABAergic and dopaminergic pathways in the central nervous system (CNS) play a pivotal role in the control of emotions and in the adaptive responses to stressful stimuli. The present study was aimed at characterizing a range of biochemical markers of GABA- and dopamine-mediated neurotransmission in the CNS of Roman high-avoidance (RHA/Verh) and Roman low-avoidance (RLA/Verh) rats, two psychogenetically selected lines that differ in their level of emotionality. The stimulatory effect of GABA on ³⁶Cl⁻ uptake was less pronounced in the cerebral cortex of RLA/Verh rats as compared to RHA/Verh rats, whereas no line-related changes were detected in [³H]GABA and [³H]flunitrazepam binding. On the other hand, the density of D₁ dopamine receptors labeled with [³H]SCH 23390 was lower in the nucleus accumbens of RLA/Verh rats as compared to their RHA/Verh counterparts, whilst no line-dependent changes were observed in the binding parameters of D₁ dopamine receptors in the striatum, amygdala, and prefrontal cortex. These biochemical differences may contribute to the distinct emotionality and responsiveness to the effects of psychoactive drugs of RHA/Verh and RLA/Verh rats.     

Vasopressinergic modulation of stress responses in the central amygdala of the Roman high-avoidance and low-avoidance rat.

The central nucleus of the amygdala (CEA) is selectively involved in the passive component of the behavioral (immobility) and the accompanying parasympathetic response during conditioned, stressful environmental challenges. Vasopressinergic mechanisms in the brain seem to play a role in these stress responses. The effects of the neuropeptides arginine-8-vasopressin (AVP) and oxytocin (OXT) on modulating CEA activity during conditioned stress of inescapable footshock were studied in male Roman high-avoidance (RHA/Verh) and low-avoidance (RLA/Verh) rats, psychogenetically selected on the basis of shuttle-box acquisition behavior. In RLA/Verh rats, the cardiac and behavioral responses to the conditioned emotional stressor were bradycardia and immobility, suggesting an important role for the CEA in these rats. The RHA/Verh rats, however, failed to show any change in heart rate or immobility in response to a conditioned stress situation. The low dose of AVP (20 pg) in the CEA of conscious RLA/Verh rats caused an enhancement of the stress-induced bradycardiac and immobility response. However, the high dose of AVP (2 ng) and OXT (200 pg) attenuated the bradycardiac and immobility responses in the RLA/Verh rats. Infusion of AVP and OXT in the RHA/Verh rats failed to induce any change in heart rate or immobility. Binding studies revealed that the AVP receptor selectively binds AVP with high affinity. In contrast, the OXT receptor recognizes both AVP and OXT with a similar (but lower) affinity. This suggests that the behavioral and autonomic responses of the high dose of AVP may be caused by OXT receptor stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)     

The pineal complex in Roman high avoidance and Roman low avoidance rats

Summary Previous studies have shown that the pineal gland of Roman high avoidance (RHA/Verh) rats is larger than that of Roman low avoidance rats (RLA/Verh). In the present study measurement of enzyme activities (serotonin-N-acetyl-transferase, hydroxyindole-O-methyltransferase) revealed that pineals of RHA/Verh rats are twice as active in melatonin production than pineals of RLA/Verh rats. Indoleamine content was also higher in RHA/Verh rats, whereas noradrenaline content was the same in both lines. When values were expressed per mg protein, these differences disappeared except for N-acetyl-serotonin and noradrenaline which were higher or lower in RHA/Verh rats, respectively. Both lines had higher serum levels of melatonin during the dark phase than during the light phase. However, RHA/Verh rats had increased serum levels as compared to RLA/Verh rats during both day and night. Morphometric analysis of the deep and superficial part of the pineal complex revealed, that the volumes of both parts are enlarged in RHA/Verh rats. Electron microscopic studies of pineals collected during day- and nighttime showed higher numbers of synaptic ribbons per unit area in pineals of RHA/Verh rats. In pineals collected during June synaptic ribbons displayed a day/night rhythm in RHA/Verh rats only, whereas in glands of both lines collected during November no daily changes were found. These results show that closely related but divergently selected rat lines may differ in pineal ultrastructure and pineal function.     

Stressful environmental stimuli increase extracellular DOPAC levels in the prefrontal cortex of hypoemotional (Roman high-avoidance) but not hyperemotional (Roman low-avoidance) rats. An in vivo voltammetric study

The effects of a variety of stressful environmental situations on dopamine metabolism in the prefrontal cortex (as assessed by in vivo voltammetry with carbon fiber electrodes) have been compared in two genetically selected lines of rat (Roman high (RHA/Verh) and low (RLA/Verh) avoidance) which differ drastically in their level of emotionality. Heart rate was continuously monitored in these animals (via chronically implanted subcutaneous electrodes) so as to index the emotional reaction to the stressors. An electrochemical signal corresponding to the oxidation of dihydroxyphenylacetic acid (DOPAC) was recorded in the deeper laminae of the anteromedial prefrontal cortex in both lines of rats. Under normal conditions, this signal was stable for at least 4 h and its amplitude was similar in both lines. Introduction of the animals into an unfamiliar environment (30 min), application of a mild tail pinch (10 min) or of a high-intensity loud noise (30 min) or immobilization (20 min) were all associated with an increase in extracellular cortical DOPAC levels in the hypoemotional RHA/Verh line but not in the hyperemotional RLA/Verh line. Similarly, forced locomotion on a rotarod (40 min) provoked a dramatic increase in the amplitude of the cortical DOPAC oxidation peak in RHA/Verh rats and only a mild increase in this parameter in RLA/Verh rats. In RHA/Verh rats, tolerance to this increase was observed when animals were subjected to forced locomotion every day for 5 days. All of the stressful situations investigated provoked an immediate augmentation of heart rate which resumed gradually after cessation of the stressful stimulus; the magnitude and duration of this increase were much greater in RLA/Verh than in RHA/Verh rats. Moreover, in all stress situations, RLA/Verh but not RHA/Verh rats showed behavioral signs of emotional response e.g. defecation, freezing and self-grooming. It is concluded that the increase in cortical dopamine metabolism induced by stress is not connected to the emotional reaction caused by the aversive nature of the stressor but may rather reflect a heightened attention of the animal or activation of cognitive processes in an attempt to cope with the stressor.     

Differential effects of cocaine on extracellular signal‐regulated kinase phosphorylation in nuclei of the extended amygdala and prefrontal cortex of psychogenetically selected roman high‐ and low‐avoidance rats

Roman high (RHA)‐ and low (RLA)‐avoidance rats are selectively bred for rapid vs. poor acquisition of active avoidance, respectively, and differ markedly in emotional reactivity, coping style, and behavioral and neurochemical responses to morphine and psychostimulants. Accordingly, acute cocaine induces more robust increments in locomotion and dopamine output in the nucleus accumbens shell (AcbSh) of RHA than of RLA rats. Cocaine induces short‐ and long‐term neuronal plasticity via activation of the extracellular signal‐regulated kinase (ERK) pathway. This study compares the effects of acute cocaine on ERK phosphorylation (pERK) in limbic brain areas of Roman rats. In RHA but not RLA rats, cocaine (5 mg/kg) increased pERK in the infralimbic prefrontal cortex and AcbSh, two areas involved in its acute effects, but did not modify pERK in the prelimbic prefrontal cortex and Acb core, which mediate the chronic effects of cocaine. Moreover, cocaine failed to affect pERK immunolabeling in the bed nucleus of stria terminalis pars lateralis and central amygdala of either line but increased it in the basolateral amygdala of RLA rats. These results extend to pERK expression previous findings on the greater sensitivity to acute cocaine of RHA vs. RLA rats and confirm the notion that genetic factors influence the differential responses of the Roman lines to addictive drugs. Moreover, they support the view that the Roman lines are a useful tool to investigate the molecular underpinnings of individual vulnerability to drug addiction. © 2014 Wiley Periodicals, Inc.     

Differential hypothalamic self-stimulation behaviour in roman high-avoidance and low-avoidance rats

Monopolar electrodes were implanted bilaterally and symmetrically into a small well defined region of the posterior lateral hypothalamus in 11 rats of the selected strains Roman high-avoidance (RHA/Verh, 6 animals) and Roman low-avoidance (RLA/Verh, 5 animals). Intracranial electrical stimulation (ICS) was used to study possible strain difference with respect to self-stimulation behaviour. Using a shuttle-box, the experimental set-up allowed measurements of the animal's preference, aversion, neutrality or ambivalence towards the stimulus. For both rat strains 10 electrode sites were tested with identical stimulation currents (100–600 μA in increments of 100 μA). Significant differences between the selected strains were found for preference (self-stimulation) and aversion: RLA/Verhrats showed self-stimulation at lower stimulation currents than RHA/Verh-rats. At higher currents, both strains showed similar self-stimulation performance. RLA/Verh-rats escaped more often from ICS yielding self-stimulation at five of the six current levels. It is concluded that, in comparison with the RHA strain, RLA/Verh-rats are more sensitive to aversive effects of lateral hypothalamic stimulation. This seems to be associated with an increased sensitivity to the rewarding effects of such stimulation.     

Infusion of flesinoxan into the amygdala blocks the fear-potentiated startle

In a previous study it was demonstrated that flesinoxan, a selective serotonin (5-HT)1A receptor agonist, had anxiolytic properties in the fear-potentiated startle paradigm. The present study investigated the putative site of action of flesinoxan in this paradigm. Flesinoxan infused either into the dorsal raphe nucleus or the median raphe nucleus did not affect startle potentiation. Bilateral infusion of flesinoxan into the central nucleus of the amygdala on the other hand, dose-dependently blocked the fear-potentiated startle response. These data indicate that flesinoxan exerts it anxiolytic effects in the fear-potentiated startle paradigm via the central nucleus of the amygdala, whereas the dorsal and median raphe nuclei are not directly involved in this process.     

Brain Metabolism of Progesterone, Coping Behaviour and Emotional Reactivity in Male Rats from Two Psychogenetically Selected Lines

Brain metabolites of progesterone such as tetrahydroprogesterone (THP) act on GABA_A receptors and have anxiolytic properties. The formation of THP and its 5α-reduced precursor, dihydroprogestrone (DHP) was measured in vitro in various microdissected brain areas obtained from males of two psychogenetically selected rat lines, i.e. the Roman High-(RHA/Verh) and Low-(RLA/Verh) Avoidance rats, which are known to differ in emotional reactivity and/or anxiety. The behavioural and neuroendocrine responses of these rats were also measured following exposure to a novel environment in two different test situations. The formation of DHP and THP was found to be significantly higher in the frontal cortex (FCX), and DHP in the bed nucleus of the stria terminalis (BST), of the hypoemotional RHA/Verh rats. In addition, enzymatic activity in the FCX was found to be inversely correlated with behavioural measures of anxiety. These results suggest that individual, possibly genetically-determined differences in brain production of endogenous anxiolytics derived from progesterone may account at least in part for the behavioural differences characterizing these two lines, and provide further evidence that neurosteroids acting on the GABAergic system may play an important role in modulating physiological and/or behavioural responses to environmental stressors.     

Anxiolytic effects of kindling role of anatomical location of the kindling electrode in response to kindling of the right basolateral amygdala

Study of effects of kindling on affect has been complicated by the fact that anxiogenic, anxiolytic or no effects may be observed following kindling of the amygdala. Factors affecting behavioral outcome include strain of rat, hemisphere kindled, amygdala nucleus kindled and location of the kindling electrodes within particular AP planes of a given nucleus. Previous work has suggested that kindling of the right basolateral amygdala (BLA) is predominantly anxiogenic. This conclusion was based on kindling of anterior or posterior parts of the BLA. The present study sought to clarify this conclusion by examining behavioral effects of right BLA kindling in a mid-range of AP planes not yet studied. A variety of measures of rodent anxiety-like behavior were examined, including behavior in the hole board, elevated plus maze, light/dark box, social interaction test and unconditioned acoustic startle. Anhedonic effects of kindling were assessed by a sucrose preference test with controls for fluid consumption and taste sensitivities. All effects were assessed shortly after kindling (1-2 days) and at a longer time interval (7-8 days). Kindling to four stage 5 seizures in the mid-right BLA altered behavior at all time points after kindling in all tests except the hole board and light/dark box tests. The effect of kindling was anxiolytic like in the plus maze, social interaction and startle tests. Kindling in mid-BLA also increased sucrose consumption. Effects on sucrose consumption are consistent with previous studies showing no depressive-like effects of amygdala kindling in rodents. It is hypothesized that the focal nature of the behavioral consequences of amygdala kindling are best understood in the context of the circuitry in which the cells stimulated are imbedded and the impact of kindling on functioning of those circuits.     

Corticotropin-releasing Factor in the Caudal Pontine Reticular Nucleus Mediates the Expression of Fear-potentiated Startle in the Rat

The fear-potentiated startle paradigm is a valuable model for the investigation of the neuronal basis of fear. Previous studies have demonstrated that the neuropeptide corticotropin-releasing factor (CRF) plays an important role in fear-related processes, notably in the potentiation of the acoustic startle response. The present study investigated the role in fear-potentiated startle of CRF in the caudal pontine reticular nucleus, a brain nucleus that mediates the acoustic startle response. First, we showed that the central nucleus of the amygdala gives rise to a CRFergic projection to the caudal pontine reticular nucleus. In the second experiment, we iontophoretically applied CRF to caudal pontine reticular nucleus neurons and extracellularly recorded the activity of these neurons. CRF had a mainly excitatory effect on the tone-evoked activity of the neurons. In our third experiment, we injected the CRF antagonist α-helical CRF into the caudal pontine reticular nucleus of awake rats. Here, α-helical CRF dose-dependently blocked fear-potentiated startle, but had no effect on the baseline startle amplitude. The present results show that CRF-containing neurons which project from the central nucleus to the caudal pontine reticular nucleus are important for the enhancement of startle by fear, and further characterize the hypothetical neuronal circuitry underlying the expression of fear-potentiated startle.     

Anxiolytic and anxiogenic effects of kindling--role of baseline anxiety and anatomical location of the kindling electrode in response to kindling of the right and left basolateral amygdala

Effects of kindling of right and left basolateral amygdala (BLA) on plus maze anxiety was studied. Using a validated retest paradigm, it was possible to retest rats in the plus maze without increasing anxiety on retest. This permitted determining prekindling baseline levels of plus maze anxiety. Right BLA kindling of high baseline anxiety rats was anxiolytic one week after kindling. Right BLA kindling of low baseline anxiety rats was anxiogenic. In addition, left BLA kindling was either anxiogenic or without effect on plus maze anxiety, depending on baseline anxiety. Effects in left BLA differ from previous work showing anxiolytic effects of left BLA kindling. The discrepancy could be explained in part by prekindling baseline anxiety. These findings require modification of the previous conclusion that left hemisphere (left BLA) kindling is anxiolytic and right BLA kindling is anxiogenic in the plus maze. Rather the hemisphere difference may be due to an interaction between baseline anxiety level and kindling. If true, anxious disposition in rodents may interact with amygdala kindling to change amygdala function differently. Kindling and baseline anxiety effects on other behaviors (such as risk assessment and resistance to capture) are also described. Present data in the light of past studies suggest both premorbid anxiety state and location of the kindling electrode contribute to the effects of kindling on behavior.     

Hippocampal and septal injections of nicotine and 8-OH-DPAT distinguish among different animal tests of anxiety

1. Different animal tests model different anxiety disorders. Thus, the social interaction test is a model of generalised anxiety disorder, plus-maze Trial 1 models elements of panic disorder and Trial 2 in the elevated plus-maze is a model of specific phobia. 2. Studies of the neuroanatomical and neurochemical pathways controlling behaviour in these different tests provides information on the neurobiological mechanisms modulating anxiety disorders. 3. In the social interaction test, nicotine and 8-OH-DPAT had anxiogenic effects when injected into the dorsal hippocampus or the lateral septum. 4. These ligands were without effect on Trial 1 in the plus-maze when injected into the dorsal hippocampus, but had anxiogenic effects when injected into the lateral septum. 5. On Trial 2 in the elevated plus-maze, nicotine had an anxiolytic effect, but 8-OH-DPAT had an anixiogenic effect when injected into the dorsal hippocampus.     

Acoustic startle and fear-potentiated startle in rats selectively bred for fast and slow kindling rates: relation to monoamine activity

The acoustic startle response, prepulse inhibition, fear-potentiated startle and monoamine activity induced by either, a novel stimulus or a cue previously paired with foot-shock (fear-conditioning), were assessed in rats selectively bred for differences in amygdala excitability (Fast vs. Slow kindling epileptogenesis). Comorbid differences of anxiety, which were dependent both on the rats' behavioural style and the kind of stressor, also characterized these strains. In the present investigation, Slow rats exhibited a greater startle reflex to noise relative to Fast rats, suggesting differences in generalized anxiety, but similar rates of startle habituation and prepulse inhibition. The fear-potentiated startle, however, was greater in Fast rats. When movement of the rat was restricted in a new environment, presentation of a novel stimulus (light) increased norepinephrine, dopamine and/or serotonin activity in brain regions typically associated with stressors (e.g. locus coeruleus, paraventricular hypothalamic nucleus). Generally, these effects were more pronounced in Fast rats, and norepinephrine utilization in the central amygdala was particularly highlighted in response to a conditioned fear stimulus. Thus, while generalized anxiety appeared greater in Slow rats, behavioural and central neurochemical reactivity in response to novel stimuli and to fear-eliciting stimuli, was greater in Fast rats. Similarly, basal dopamine activity in the prefrontal cortex was greater in Fast rats, but dopamine utilization elicited by a novel stimulus was more pronounced in Slow rats. This suggested that relative to Slow rats, dopamine neurons in prefrontal cortex of Fast rats do not react normally to environmental stimuli, and this phenomenon could lead to disturbances of attention or impulsivity.     

Intra-amygdaloid microinjection of neuropeptide glutamic acid-isoleucine induces anxiety-like behavior

The amygdaloid complex is involved in anxiety or fear responses to stressful stimuli. In this study the effect of neuropeptide-EI on anxiety-like behavior and its influence on adrenocortical function was tested in male Wistar rats that were injected bilaterally in the basolateral amygdala with neuropeptide-EI (1 μg/1 μl) or artificial cerebrospinal fluid and placed on the plus maze. The plasma corticosterone levels were analyzed in controls and plus-maze exposed animals. Neuropeptide-EI in the basolateral amygdala significantly decreased the time spent in open arms but had no effect on locomotor activity, showing an anxiogenic effect. However, neuropeptide administration did not change serum corticosterone levels compared with vehicle controls. Our results suggest that the anxiogenic effect of neuropeptide-EI could be independent of the hypothalamic-pituitary-adrenocortical system response.     

Reduced number of CRF-containing neurons in the central amygdala correlated with enhanced locomotor activity following early postnatal corticosterone treatment in the Wistar rat

In the present study, newborn rats were implanted with corticosterone (CORT) containing polymers at postnatal day 0 (releasing rate 320-80 microg CORT/kg body weight and day). Controls received a CORT-free implant. All implants were removed at postnatal day 12. At the age of 16-20 weeks, these animals were tested for emotional behavior using an elevated plus-maze and fear-sensitized acoustic startle response. On the elevated plus-maze significant differences were found between hormone treated and control animals. The CORT-group demonstrated higher numbers of entries into closed arms and all arms, and the time spent in the center of the maze was significantly enhanced. Hormone-treated and control rats showed a significant fear sensitization of the acoustic startle response. However, no significant differences were observed between the two groups. The number of CRF-immunopositive neurons in the central nucleus of the amygdala was decreased after CORT treatment, whereas the number of NPY-immunopositive neurons and total number of neurons in the amygdala did not differ significantly between both groups. In conclusion, early postnatal stress induced by CORT administration in neonatal rats led to a higher locomotor activity correlated with changes in the number of CRF containing neurons in the central nucleus of the amygdala.     

Profiles of the antipanic compounds, triazolobenzodiazepines and phenelzine, in two animal tests of anxiety

Two animal tests of anxiety (the elevated plus-maze and the social interaction test) were used to investigate the effects of several antipanic agents. In the elevated plus-maze, the triazolobenzodiazepines adinazolam (2 and 5 mg/kg) and alprazolam (1 mg/kg), tested after 5 days of pretreatment, demonstrated significant anxiolytic effects, while phenelzine (9 mg/kg), after 21 days of pretreatment, demonstrated nonsignificant anxiolytic effects. In the social interaction test, the triazolobenzodiazepines generally did not produce an anxiolytic profile, and phenelzine even revealed significant anxiogenic activity. The antipanic agents therefore distinguish between the two tests of anxiety. The lack of a strong anxiolytic profile with these agents in both tests lends support to the distinction between generalized anxiety and panic disorder.     

Involvement of noradrenergic and corticoid receptors in the consolidation of the lasting anxiogenic effects of predator stress

The roles of beta-NER (beta-noradrenergic receptor), GR (glucocorticoid) and mineral corticoid receptors (MR) in the consolidation of anxiogenic effects of predator stress were studied. One minute after predator stress, different groups of rats were injected (ip) with vehicle, propranolol (beta-NER blocker, 5 and 10 mg/kg), mifepristone (RU486, GR blocker, 20 mg/kg), spironolactone (MR blocker, 50 mg/kg), propranolol (5 mg/kg) plus RU486 (20 mg/kg) or the anxiolytic, chloradiazepoxide (CPZ, 10 mg/kg). One week later, rodent anxiety was assessed in elevated plus maze, hole board, light/dark box, social interaction and acoustic startle. Considering all tests except startle, propranolol dose dependently blocked consolidation of lasting anxiogenic effects of predator stress in all tests. GR receptor block alone was ineffective. However, GR block in combination with an ineffective dose of propranolol did blocked consolidation of predator stress effects in all tests, suggesting a synergism between beta-NER and GR. Surprisingly, MR block prevented consolidation of anxiogenic effects in all tests except the light/dark box. CPZ post stress was ineffective against the anxiogenic impact of predator stress. Study of startle was complicated by the fact that anxiogenic effects of stress on startle amplitude manifested as both an increase and a decrease in startle amplitude. Suppression of startle occurred in stressed plus vehicle injected groups handled three times prior to predator stress. In contrast, stressed plus vehicle rats handled five times prior to predator stress showed increases in startle, as did all predator stressed only groups. Mechanisms of consolidation of the different startle responses appear to differ. CPZ post stress blocked startle suppression but not enhancement of startle. Propranolol post stress had no effect on either suppression or enhancement of startle. GR block alone post stress prevented suppression of startle, but not enhancement. In contrast blocking GR and beta-NER together prevented startle enhancement. MR block also prevented startle enhancement. Effects of MR block on startle suppression were not tested. Delay of habituation to startle was found in all stressed rats. Consolidation of delay of habituation was blocked or attenuated by post stress MR block, GR plus beta-NER block and CPZ but not by post stress GR or beta-NER block alone. Taken together, present findings suggest consolidation of lasting anxiogenic effects of predator stress may share some of the same neurochemical mechanisms implicated in some forms of fear memory consolidation. Implications of these findings for the study of stress-induced changes in affect including posttraumatic stress disorder (PTSD) are discussed.     

Infusion of the dopamine D1 receptor antagonist SCH 23390 into the amygdala blocks fear expression in a potentiated startle paradigm

Dopamine (DA) D₁ receptors are distributed in the nucleus accumbens and the amygdala, two regions of the mesocorticolimbic DA system known to be activated by aversive environmental stimuli. The objective of the present study was to determine the contribution of D₁ receptors in these brain regions to the expression of a fear-motivated behavior, notably, potentiated startle in rats. Injection of the DA D₁ receptor antagonist SCH 23390 into the amygdala blocked the ability of a conditioned light stimulus previously paired with footshock to enhance acoustic startle amplitudes. Bilateral intracerebral administration of SCH 23390 into the nucleus accumbens had no effect on fear-potentiated startle. The observed opposing effects of amygdaloid DA D₁ receptor antagonism on fear expression, along with earlier research demonstrating the involvement of ventral tegmental area (VTA) DA neurons on fear-potentiated startle, suggest a role for mesoamygdaloid activity in conditioned excitatory fear reactions.