Differential effects of basolateral amygdala lesions on behavior, corticosterone, and prolactin responses

The present experiment examines hormone-behavior relationships following manipulation of the amygdala. Affective behavior and levels of corticosterone and prolactin were compared in rats with lesions of the basolateral amygdala and in nonlesioned and sham-operated controls. Animals with lesions of the basolateral amygdala were found to be hyperreactive and to have normal resting levels of corticosterone and prolactin but potentiated corticosterone responses to stress. Normal prolactin stress responses were unaltered by the lesion. The results are discussed in relation to behavioral and endocrine changes seen following other limbic system lesions.     

Input-specific LTP and depotentiation in the basolateral amygdala

The amygdala plays a central role in emotional memory. The cellular mechanisms by which the amygdala participates in emotional learning are believed to be changes in efficacy of synaptic transmission, similar to long-term potentiation (LTP) and long-term depression (LTD). Although different forms of LTP have been shown in the amygdala, many of their features are still unknown. Here, we use both field potential and intracellular recordings in rat amygdala slices, and show that LTP in the basolateral nucleus, induced by high-frequency stimulation (HFS) of the external capsule is input-specific, can be reversed by low-frequency stimulation (LFS), and can be reinstated by HFS. These synapse-specific, reversible changes in synaptic strength in the basolateral nucleus of the amygdala may be important to amygdala's role in emotional memory.     

Excitatory amino acid receptors in the basolateral amygdala regulate anxiety responses in the social interaction test

Blocking GABA_A receptors in the basolateral amygdala (BLA) elicits increases in heart rate (HR), blood pressure (BP) and anxiety responses by enhancing a glutamate mediated excitation. The present study was conducted to determine the role of the ionotropic glutamate receptors within the BLA in regulating HR, BP and experimental anxiety. Blocking basal glutamate excitation had no significant effect on HR or BP, but did elicit a significant anxiolytic-like effect.     

The basolateral amygdala in reward learning and addiction

Sophisticated behavioral paradigms partnered with the emergence of increasingly selective techniques to target the basolateral amygdala (BLA) have resulted in an enhanced understanding of the role of this nucleus in learning and using reward information. Due to the wide variety of behavioral approaches many questions remain on the circumscribed role of BLA in appetitive behavior. In this review, we integrate conclusions of BLA function in reward-related behavior using traditional interference techniques (lesion, pharmacological inactivation) with those using newer methodological approaches in experimental animals that allow in vivo manipulation of cell type-specific populations and neural recordings. Secondly, from a review of appetitive behavioral tasks in rodents and monkeys and recent computational models of reward procurement, we derive evidence for BLA as a neural integrator of reward value, history, and cost parameters. Taken together, BLA codes specific and temporally dynamic outcome representations in a distributed network to orchestrate adaptive responses. We provide evidence that experiences with opiates and psychostimulants alter these outcome representations in BLA, resulting in long-term modified action.     

Cholecystokinin enhances GABAergic inhibitory transmission in basolateral amygdala

The neuropeptide cholecystokinin (CCK) is anxiogenic in studies of human and animal behavior. As the amygdala formation has been implicated in generation of emotional states such as anxiety, we tested the effect of CCK on spontaneous synaptic events in the basolateral amygdala (BLA) using whole cell patch recordings in rat brain slice preparation. We found that CCK increased the frequency of spontaneous inhibitory postsynaptic potentials (sIPSPs) and currents (sIPSCs). This effect was blocked by the fast sodium channel blocker tetrodotoxin (TTX), indicating that the CCK effect is likely mediated by direct excitation of GABAergic interneurons. The CCK(B) receptor subtype antagonist, CR2945, blocked the CCK effect, while CCK4, a specific CCK(B) agonist, increased sIPSC frequency. We hypothesize that these actions may underlie the anxiogenic effects of CCK observed in behavioral studies.     

Role of corticotropin-releasing factor and urocortin within the basolateral amygdala of rats in anxiety and panic responses

The amygdala is a critical temporal lobe structure involved in the expression of anxiety and stress responses. The basolateral nucleus (BLA) of the amygdala in particular, may play a key role in anxiety. Furthermore, corticotropin-releasing factor (CRF), a 41 amino acid peptide, has been strongly implicated in the regulation of stress and anxiety responses. Centrally administered CRF has been shown to increase the anxiety-like behaviors of rodents in several animal models. A recently cloned related peptide, Urocortin (Ucn), appears to have similar affinity for the CRF₁ receptor, but higher affinity at the CRF₂ receptor. When microinjected into the BLA, we found Ucn was substantially more potent than CRF in producing anxiogenic-like behavior as assessed in the social interaction test. Furthermore, repetitive administration of subthreshold doses of Ucn and CRF resulted in ‘priming'. Once primed, these animals exhibited behavioral and cardiovascular responses to intravenous sodium lactate, a panicogenic agent in susceptible human patients. These results suggest central CRF and Ucn play a role in generating anxiety which may be similar to that seen in pathological conditions such as panic disorder.     

Hypervigilance for fear after basolateral amygdala damage in humans

Recent rodent research has shown that the basolateral amygdala (BLA) inhibits unconditioned, or innate, fear. It is, however, unknown whether the BLA acts in similar ways in humans. In a group of five subjects with a rare genetic syndrome, that is, Urbach–Wiethe disease (UWD), we used a combination of structural and functional neuroimaging, and established focal, bilateral BLA damage, while other amygdala sub-regions are functionally intact. We tested the translational hypothesis that these BLA-damaged UWD-subjects are hypervigilant to facial expressions of fear, which are prototypical innate threat cues in humans. Our data indeed repeatedly confirm fear hypervigilance in these UWD subjects. They show hypervigilant responses to unconsciously presented fearful faces in a modified Stroop task. They attend longer to the eyes of dynamically displayed fearful faces in an eye-tracked emotion recognition task, and in that task recognize facial fear significantly better than control subjects. These findings provide the first direct evidence in humans in support of an inhibitory function of the BLA on the brain''s threat vigilance system, which has important implications for the understanding of the amygdala''s role in the disorders of fear and anxiety.     

Ipsilateral trigeminal sensory responses to cortical stimulation by subdural electrodes

Twelve patients with medically intractable epilepsy had plates of chronic subdural electrodes placed over the lateral and basal cortical hemispheres during evaluations for surgical therapy. During cortical stimulation, ipsilateral sensations involving any of the branches of the trigeminal nerve were noted in the eye, face, and mouth. Some responses could have been due to dural or direct trigeminal nerve trunk stimulation, but others were probably due to electrical stimulation of trigeminal fibers accompanying the pial-arachnoidal vessels. These fibers had been demonstrated in animals, but not in humans.     

Photoperiod alters fear responses and basolateral amygdala neuronal spine density in white-footed mice (Peromyscus leucopus)

Photoperiodism is a biological phenomenon in which environmental day length is monitored to ascertain time of year to engage in seasonally appropriate adaptations. This trait is common among organisms living outside of the tropics. White-footed mice (Peromyscus leucopus) are small photoperiodic rodents which display a suite of adaptive responses to short day lengths, including reduced hippocampal volume, impairments in hippocampal-mediated memory, and enhanced hypothalamic-pituitary-adrenal axis reactivity. Because these photoperiodic changes in brain and behavior mirror some of the etiology of post-traumatic stress disorder (PTSD), we hypothesized that photoperiod may also alter fear memory and neuronal morphology within the hippocampus-basolateral amygdala-prefrontal cortex fear circuit. Ten weeks of exposure to short days increased fear memory in an auditory-cued fear conditioning test. Short days also increased dendritic spine density of the neurons of the basolateral amygdala, without affecting morphology of pyramidal neurons within the infralimbic region of the medial prefrontal cortex. Taken together, photoperiodic phenotypic changes in brain morphology and physiology induced by a single environmental factor, exposure to short day lengths, affect responses to fearful stimuli in white-footed mice. These results have potential implications for understanding seasonal changes in fear responsiveness, as well as for expanding translational animal models for studying gene-environment interactions underlying psychiatric diseases, such as PTSD.     

Postsynaptic targets of somatostatin-containing interneurons in the rat basolateral amygdala

The basolateral amygdala contains several subpopulations of inhibitory interneurons that can be distinguished on the basis of their content of calcium-binding proteins or peptides. Although previous studies have shown that interneuronal subpopulations containing parvalbumin (PV) or vasoactive intestinal peptide (VIP) innervate distinct postsynaptic domains of pyramidal cells as well as other interneurons, very little is known about the synaptic outputs of the interneuronal subpopulation that expresses somatostatin (SOM). The present study utilized dual-labeling immunocytochemical techniques at the light and electron microscopic levels to analyze the innervation of pyramidal cells, PV+ interneurons, and VIP+ interneurons in the anterior basolateral amygdalar nucleus (BLa) by SOM+ axon terminals. Pyramidal cell somata and dendrites were selectively labeled with antibodies to calcium/calmodulin-dependent protein kinase II (CaMK); previous studies have shown that the vast majority of dendritic spines, whether CAMK+ or not, arise from pyramidal cells. Almost all SOM+ axon terminals formed symmetrical synapses. The main postsynaptic targets of SOM+ terminals were small-caliber CaMK+ dendrites and dendritic spines, some of which were CaMK+. These SOM+ synapses with dendrites were often in close proximity to asymmetrical (excitatory) synapses to these same structures formed by unlabeled terminals. Few SOM+ terminals formed synapses with CaMK+ pyramidal cell somata or large-caliber (proximal) dendrites. Likewise, only 15% of SOM+ terminals formed synapses with PV+, VIP+, or SOM+ interneurons. These findings suggest that inhibitory inputs from SOM+ interneurons may interact with excitatory inputs to pyramidal cell distal dendrites in the BLa. These interactions might affect synaptic plasticity related to emotional learning.     

Single neuron responses in amygdala of alert monkey during complex sensory stimulation with affective significance

Among other deficits, amygdalectomy impairs the ability of the animal to recognize the affective significance of a stimulus. In the present study, neuronal activity in the amygdala (AM) was recorded from alert monkeys while they performed tasks leading to the presentation of rewarding or aversive stimuli. Of 585 AM neurons tested, 312 (53.3%) responded to at least one stimulus in one or more of 5 major groups: 40 vision related, 26 audition related, 41 ingestion related, 117 multimodal, and 14 selective. Ingestion-related neurons were subdivided according to their responses to other stimuli: oral sensory, oral sensory plus vision, and oral sensory plus audition. Depending upon their responsiveness to the affective significance of the stimuli, neurons in the vision- and audition-related categories were divided into 2 subclasses: vis-I (26/40), vis-II (14/40), aud-I (8/26), and aud-II (18/26). All 4 subtypes usually responded to unfamiliar stimuli but seldom responded to neutral familiar stimuli. Types vis-I and aud-I responded to both positive and negative familiar stimuli. Types vis-II and aud-II responded to certain familiar negative stimuli but not to familiar positive stimuli. In vis-I neurons, responses were stronger for palatable foods than for less palatable foods. No neurons within vision-related, audition-related, and multimodal categories responded solely to positive or to negative stimuli. Of the 27 oral sensory neurons 9 were tested with saline or salted food, and 8 responded to normally aversive oral sensory stimuli in the same manner as they did to normal food or liquid (water or juice). In contrast to oral sensory neurons, all responses of 4 oral sensory-plus-vision and all of 4 selective neurons tested, as well as bar pressing behavior, were modulated by altering the affective significance of the food. These results suggest that the AM is one of the candidates for stimulus-affective association based on associative learning and memory.     

Increased amphetamine-induced locomotion during inactivation of the basolateral amygdala

At low doses, amphetamine has been shown to produce reliable increases in locomotor activity through its actions on the mesolimbic dopamine (DA) terminals in the nucleus accumbens (NAC). The basolateral amygdala (BLA) has recently been reported to have anatomical projections to the NAC, suggesting that it might serve to alter or modulate the function of the NAC. To test this hypothesis, the current experiment produced lidocaine-reversible lesions of the BLA and assessed changes in NAC function by examining alterations in locomotor activity in response to s.c. amphetamine (2mg/kg). While BLA inactivation alone was found to have no effect on spontaneous or basal locomotor activity, it produced a significant potentiation of amphetamine-induced hyperactivity. These results suggest that BLA inactivation removes a system that inhibits the locomotor response to amphetamine. The data are, therefore, consistent with the view that the BLA may serve to modulate NAC function.     

Differential roles of the prefrontal cortical subregions and basolateral amygdala in compulsive cocaine seeking and relapse after voluntary abstinence in rats

Compulsive drug use and a persistent vulnerability to relapse are key features of addiction. Imaging studies have suggested that these features may result from deficits in prefrontal cortical structure and function, and thereby impaired top‐down inhibitory control over limbic–striatal mechanisms of drug‐seeking behaviour. We tested the hypothesis that selective damage to distinct subregions of the prefrontal cortex, or to the amygdala, after a short history of cocaine taking would: (i) result in compulsive cocaine seeking at a time when it would not usually be displayed; or (ii) facilitate relapse to drug seeking after abstinence. Rats with selective, bilateral excitotoxic lesions of the basolateral amygdala or anterior cingulate, prelimbic, infralimbic, orbitofrontal or anterior insular cortices were trained to self‐administer cocaine under a seeking–taking chained schedule. Intermittent mild footshock punishment of the cocaine‐seeking response was then introduced. No prefrontal cortical lesion affected the ability of rats to withhold their seeking responses. However, rats with lesions to the basolateral amygdala increased their cocaine‐seeking responses under punishment and were impaired in their acquisition of conditioned fear. Following a 7‐day abstinence period, rats were re‐exposed to the drug‐seeking environment for assessment of relapse in the absence of punishment or cocaine. Rats with prelimbic cortex lesions showed decreased seeking responses during relapse, whereas those with anterior insular cortex lesions showed an increase. Combined, these results show that acute impairment of prefrontal cortical function does not result in compulsive cocaine seeking after a short history of self‐administering cocaine, but further implicates subregions of the prefrontal cortex in relapse.     

Changes in cortical sensory responses with a cryogenic blockade of nucleus ventralis posterolateralis

The contribution of nucleus ventralis posterolateralis (VPL) to sensory responses in the cortex of the cat was investigated. Averaged evoked potentials (AEP) were collected in response to paw, click, and light stimuli from primary sensory area S1 and the precruciate (PCA), anterior lateral (ALA), and posterior middle suprasylvian (PMSA) cortical association areas. Activity in VPL was reversibly blocked using a cryoprobe system to produce localized cooling. Reduction in amplitude of the short latency component of the AEP in S1 in response to the paw stimulus indicated that VPL was blocked. The short latency component of responses in the three association cortices was also blocked by cooling VPL, but the latency of this component was 20 msec longer than that of S1. Cross-correlation analyses of AEPs were performed to evaluate waveform similarity. Responses in a single cortical area to two different stimulus modalities or responses in two different cortical areas to one stimulus modality had about the same degree of waveform similarity. The effect of cooling VPL on waveform similarity was minimal and was not consistent for either stimulus pairs or cortical pairs. We concluded that VPL contributes to short latency responses in association cortices, but by a projection system differing from that to S1 since the latency to the changed response component was 20 msec longer than that observed for S1 cortex. Some similarity of sensory response waveform was found, but VPL makes little or no contribution to this similarity. No evidence was found for covariation of response excitability in cortical areas.     

Relaxin receptor activation in the basolateral amygdala impairs memory consolidation

The peptide-hormone relaxin has well-established actions in male and female reproductive tracts, and has functional effects in circumventricular regions of brain involved in neurohormonal secretion. In the current study, we initially mapped the distribution of mRNA encoding the relaxin receptor--leucine-rich repeat-containing G-protein-coupled receptor 7 (LGR7)- and [33P]-human relaxin-binding sites in extra-hypothalamic sites of male Sprague-Dawley rats. The basolateral amygdala (BLA) expressed high levels of LGR7 mRNA and relaxin-binding sites and, although relaxin peptide was not detected in the BLA, several brain regions that send projections to the BLA were found to contain relaxin-expressing neurons. As it is well established that the BLA is involved in regulating the consolidation of memory for emotionally arousing experiences, we investigated whether activation of LGR7 in the BLA modulated memory consolidation for aversively motivated inhibitory avoidance training. Bilateral infusions of human relaxin (10-200 ng in 0.2 microL) into the BLA immediately after inhibitory avoidance training impaired 48-h retention performance in a dose-dependent manner. Delayed infusions of relaxin into the BLA 3 h after training were ineffective, indicating that the retention impairment was due to influences on memory consolidation. Post-training infusions of relaxin into the adjacent central amygdala, which is devoid of LGR7, did not impair retention. These findings suggest a novel function for endogenous relaxin-LGR7 signalling in rat brain involving regulation of memory consolidation.     

Blockade of noradrenergic receptors in the basolateral amygdala impairs taste memory

In conditioned taste aversion (CTA), a subject learns to associate a novel taste (conditioned stimulus, CS) with visceral malaise (unconditioned stimulus, US). Considerable evidence indicates that the noradrenergic system in the amygdala plays an important role in memory consolidation for emotionally arousing experiences. The specific aim of the present set of experiments was to determine the involvement of noradrenergic activity in the basolateral amygdala (BLA) during the US presentation and consolidation of CTA as well as during the consolidation of a nonaversive/incidental gustatory memory. Selective bilateral microinfusions of the beta-adrenergic antagonist propranolol administered into the BLA immediately before intraperitoneal (i.p.) lithium chloride (LiCl) injections disrupted CTA memory. Additionally, propranolol infused into the BLA immediately after a pre-exposure to the saccharin (CS) significantly attenuated latent inhibition. The present findings indicating that alterations in noradrenergic function in the BLA affect taste memory formation, provide additional evidence that the BLA plays a critical role in modulating the consolidation of memory and that the influence is mediated by interactions with other brain regions that support memory for different kinds of experiences.