Approaches to characterizing oscillatory burst detection algorithms for electrophysiological recordings.

BACKGROUND: Cognitive processes are associated with fast oscillations of the local field potential and electroencephalogram. There is a growing interest in targeting them because these are disrupted by aging and disease. This has proven challenging because they often occur as short-lasting bursts. Moreover, they are obscured by broad-band aperiodic activity reflecting other neural processes. These attributes have made it exceedingly difficult to develop analytical tools for estimating the reliability of detection methods. NEW METHOD: To address this challenge, we developed an open-source toolkit with four processing steps, that can be tailored to specific brain states and individuals. First, the power spectrum is decomposed into periodic and aperiodic components, each of whose properties are estimated. Second, the properties of the transient oscillatory bursts that contribute to the periodic component are derived and optimized to account for contamination from the aperiodic component. Third, using the burst properties and aperiodic power spectrum, surrogate neural signals are synthesized that match the observed signal's spectrotemporal properties. Lastly, oscillatory burst detection algorithms run on the surrogate signals are subjected to a receiver operating characteristic analysis, providing insight into their performance. RESULTS: The characterization algorithm extracted features of oscillatory bursts across multiple frequency bands and brain regions, allowing for recording-specific evaluation of detection performance. For our dataset, the optimal detection threshold for gamma bursts was found to be lower than the one commonly used. COMPARISON WITH EXISTING METHODS: Existing methods characterize the power spectrum, while ours evaluates the detection of oscillatory bursts. CONCLUSIONS: This pipeline facilitates the evaluation of thresholds for detection algorithms from individual recordings.

The amygdala mediates the facilitating influence of emotions on memory through multiple interacting mechanisms.

Emotionally arousing experiences are better remembered than neutral ones, highlighting that memory consolidation differentially promotes retention of experiences depending on their survival value. This paper reviews evidence indicating that the basolateral amygdala (BLA) mediates the facilitating influence of emotions on memory through multiple mechanisms. Emotionally arousing events, in part by triggering the release of stress hormones, cause a long-lasting enhancement in the firing rate and synchrony of BLA neurons. BLA oscillations, particularly gamma, play an important role in synchronizing the activity of BLA neurons. In addition, BLA synapses are endowed with a unique property, an elevated post-synaptic expression of NMDA receptors. As a result, the synchronized gamma-related recruitment of BLA neurons facilitates synaptic plasticity at other inputs converging on the same target neurons. Given that emotional experiences are spontaneously remembered during wake and sleep, and that REM sleep is favorable to the consolidation of emotional memories, we propose a synthesis for the various lines of evidence mentioned above: gamma-related synchronized firing of BLA cells potentiates synapses between cortical neurons that were recruited during an emotional experience, either by tagging these cells for subsequent reactivation or by enhancing the effects of reactivation itself.

The Basolateral Amygdala Sends a Mixed (GABAergic and Glutamatergic) Projection to the Mediodorsal Thalamic Nucleus.

The medial prefrontal cortex receives converging inputs from the mediodorsal thalamic nucleus (MD) and basolateral amygdala (BLA). Although many studies reported that the BLA also projects to MD, there is conflicting evidence regarding this projection, with some data suggesting that it originates from GABAergic or glutamatergic neurons. Therefore, the present study aimed to determine the neurotransmitter used by MD-projecting BLA cells in male and female rats. We first examined whether BLA cells retrogradely labeled by Fast Blue infusions in MD are immunopositive for multiple established markers of BLA interneurons. A minority of MD-projecting BLA cells expressed somatostatin (∼22%) or calretinin (∼11%) but not other interneuronal markers, suggesting that BLA neurons projecting to MD not only include glutamatergic cells, but also long-range GABAergic neurons. Second, we examined the responses of MD cells to optogenetic activation of BLA axons using whole-cell recordings in vitro Consistent with our immunohistochemical findings, among responsive MD cells, light stimuli typically elicited isolated EPSPs (73%) or IPSPs (27%) as well as coincident EPSPs and IPSPs (11%). Indicating that these IPSPs were monosynaptic, light-evoked EPSPs and IPSPs had the same latency and the IPSPs persisted in the presence of ionotropic glutamate receptor antagonists. Overall, our results indicate that the BLA sends a mixed, glutamatergic-GABAergic projection to MD, which likely influences coordination of activity between BLA, MD, and medial prefrontal cortex. An important challenge for future studies will be to examine the connections formed by MD-projecting glutamatergic and GABAergic BLA cells with each other and other populations of BLA cells.SIGNIFICANCE STATEMENT The mediodorsal thalamic nucleus (MD) and basolateral amygdala (BLA) send convergent projections to the medial prefrontal cortex. Although many studies reported that the BLA also projects to MD, there is conflicting evidence as to whether this projection is glutamatergic or GABAergic. By combining tract tracing, immunohistochemistry, optogenetics, and patch clamp recordings in vitro, we found that BLA neurons projecting to MD not only include glutamatergic cells, but also long-range GABAergic neurons. Differential recruitment of these two contingents of cells likely influences coordination of activity between the BLA, MD, and medial prefrontal cortex.

Human anterior insula signals salience and deviations from expectations via bursts of beta oscillations.

Functional imaging studies indicate that the insula encodes the salience of stimuli and deviations from expectations, signals that can mobilize cognitive resources and facilitate learning. However, there is no information about the physiological underpinnings of these phenomena beyond changing BOLD signals. To shed light on this question, we analyzed intracerebral local field potentials (LFPs) in five patients with epilepsy of both genders performing a virtual reality task that featured varying odds of monetary rewards and losses. Upon outcome disclosure, the anterior (but not the posterior) insula generated bursts of beta oscillations whose amplitudes were lower for neutral than positive and negative outcomes, consistent with a salience signal. Moreover, beta burst power was higher when outcomes deviated from expectations, whether the outcome was better or worse than expected, indicating that the insula provides an unsigned prediction error signal. Last, in relation to insular beta bursts, many higher-order cortical areas exhibited robust changes in LFP activity that ranged from spectrally nonspecific or differentiated increases in gamma power to bursts of beta activity that closely resembled the insular beta bursts themselves. Critically, the activity of these other cortical regions was more closely tied in time to insular bursts than task events, suggesting that they are associated with particularly significant cognitive phenomena. Overall, our findings suggest that the insula signals salience and prediction errors via amplitude modulations of beta bursts, which coincide with the near simultaneous recruitment of vast cortical territories.NEW & NOTEWORTHY Functional imaging studies indicate that the anterior insula encodes salience and deviations from expectations. Beyond changing BOLD signals, however, the physiological underpinnings of these signals are unknown. By recording local field potentials in patients with epilepsy, we found that the anterior insula generates large bursts of beta oscillations whose amplitude is modulated by the salience of outcomes and deviations from expectations. Moreover, insular beta bursts coincide with the activation of many high-order cortical areas.

INFLUENCE OF RAT CENTRAL THALAMIC NEURONS ON FORAGING BEHAVIOR IN A HAZARDOUS ENVIRONMENT.

Foraging entails a complex balance between approach and avoidance alongside sensorimotor and homeostatic processes under the control of multiple cortical and subcortical areas. Recently, it has become clear that several thalamic nuclei located near the midline regulate motivated behaviors. However, one midline thalamic nucleus that project to key nodes in the foraging network, the central medial (CMT) nucleus, has received little attention so far. Therefore, the present study examined CMT contributions to foraging behavior using inactivation and unit recording techniques in male rats. Inactivation of CMT or the basolateral amygdala (BLA) with muscimol abolished the rats' normally cautious behavior in the foraging task. Moreover, CMT neurons showed large but heterogeneous activity changes during the foraging task, with many neurons decreasing or increasing their discharge rates, with a modest bias for the latter. A generalized linear model revealed that the nature (inhibitory vs. excitatory) and relative magnitude of the activity modulations seen in CMT neurons differed markedly from those of principal BLA cells but were very similar to those of fast-spiking BLA interneurons. Together, these findings suggest that CMT is an important regulator of foraging behavior. In the Discussion, we consider how CMT is integrated in the network of structures that regulate foraging.SIGNIFICANCE STATEMENTForaging entails a complex balance between approach and avoidance alongside sensorimotor and homeostatic processes under the control of multiple cortical and subcortical areas. Although the central medial thalamic (CMT) nucleus is connected to many nodes in this network, its role in the regulation of foraging behavior has not been investigated so far. Here, we examined CMT contributions to foraging behavior using inactivation and unit recording techniques. We found that CMT inactivation abolishes the rats' normally cautious foraging behavior and that CMT neurons show large but heterogeneous changes in firing rates during the foraging task. Together, these results suggest that CMT is an important regulator of foraging behavior.

Optogenetic study of central medial and paraventricular thalamic projections to the basolateral amygdala.

The central medial (CMT) and paraventricular (PVT) thalamic nuclei project strongly to the basolateral amygdala (BL). Similarities between the responsiveness of CMT, PVT, and BL neurons suggest that these nuclei strongly influence BL activity. Supporting this possibility, an electron microscopic study reported that, in contrast with other extrinsic afferents, CMT and PVT axon terminals form very few synapses with BL interneurons. However, since limited sampling is a concern in electron microscopic studies, the present investigation was undertaken to compare the impact of CMT and PVT thalamic inputs on principal and local-circuit BL neurons with optogenetic methods and whole cell recordings in vitro. Optogenetic stimulation of CMT and PVT axons elicited glutamatergic excitatory postsynaptic potentials (EPSPs) or excitatory postsynaptic currents (EPSCs) in principal cells and interneurons, but they generally had a longer latency in interneurons. Moreover, after blockade of polysynaptic interactions with tetrodotoxin (TTX), a lower proportion of interneurons (50%) than principal cells (90%) remained responsive to CMT and PVT inputs. Although the presence of TTX-resistant responses in some interneurons indicates that CMT and PVT inputs directly contact some local-circuit cells, their lower incidence and amplitude after TTX suggest that CMT and PVT inputs form fewer synapses with them than with principal BL cells. Together, these results indicate that CMT and PVT inputs mainly contact principal BL neurons such that when CMT or PVT neurons fire, limited feedforward inhibition counters their excitatory influence over principal BL cells. However, CMT and PVT axons can also recruit interneurons indirectly, via the activation of principal cells, thereby generating feedback inhibition.NEW & NOTEWORTHY Midline thalamic (MTh) nuclei contribute major projections to the basolateral amygdala (BL). Similarities between the responsiveness of MTh and BL neurons suggest that MTh neurons exert a significant influence over BL activity. Using optogenetic techniques, we show that MTh inputs mainly contact principal BL neurons such that when MTh neurons fire, little feedforward inhibition counters their excitatory influence over principal cells. Thus, MTh inputs may be major determinants of BL activity.

Gamma oscillations in the basolateral amygdala: localization, microcircuitry, and behavioral correlates.

The lateral (LA) and basolateral (BL) nuclei of the amygdala regulate emotional behaviors. Despite their dissimilar extrinsic connectivity, they are often combined, perhaps because their cellular composition is similar to that of the cerebral cortex, including excitatory principal cells reciprocally connected with fast-spiking interneurons (FSIs). In the cortex, this microcircuitry produces gamma oscillations that support information processing and behavior. We tested whether this was similarly the case in the rat (males) LA and BL using extracellular recordings, biophysical modeling, and behavioral conditioning. During periods of environmental assessment, both nuclei exhibited gamma oscillations that stopped upon initiation of active behaviors. Yet, BL exhibited more robust spontaneous gamma oscillations than LA. The greater propensity of BL to generate gamma resulted from several microcircuit differences, especially the proportion of FSIs and their interconnections with principal cells. Furthermore, gamma in BL but not LA regulated the efficacy of excitatory synaptic transmission between connected neurons. Together, these results suggest fundamental differences in how LA and BL operate. Most likely, gamma in LA is externally driven whereas in BL, it can also arise spontaneously to support ruminative processing and the evaluation of complex situations.SIGNIFICANCE STATEMENT:The basolateral amygdala (BLA) participates in the production and regulation of emotional behaviors. It is thought to perform this using feedforward circuits that enhance stimuli that gain emotional significance and directs them to valence-appropriate downstream effectors. This perspective overlooks the fact that its microcircuitry is recurrent and potentially capable of generating oscillations in the gamma band (50-80 Hz), which synchronize spiking activity and modulate communication between neurons. This study found that BLA gamma supports both these processes, is associated with periods of action selection and environmental assessment irrespective of valence, and differs between BLA subnuclei in a manner consistent with their heretofore unknown microcircuit differences. Thus, it provides new mechanisms for BLA to support emotional behaviors.

Different Multidimensional Representations across the Amygdalo-Prefrontal Network during an Approach-Avoidance Task.

The prelimbic (PL) area and basolateral amygdala (lateral [LA] and basolateral [BL] nuclei) have closely related functions and similar extrinsic connectivity. Reasoning that the computational advantage of such redundancy should be reflected in differences in how these structures represent information, we compared the coding properties of PL and amygdala neurons during a task that requires rats to produce different conditioned defensive or appetitive behaviors. Rather than unambiguous regional differences in the identities of the variables encoded, we found gradients in how the same variables are represented. Whereas PL and BL neurons represented many different parameters through minor variations in firing rates, LA cells coded fewer task features with stronger changes in activity. At the population level, whereas valence could be easily distinguished from amygdala activity, PL neurons could distinguish both valence and trial identity as well as or better than amygdala neurons. Thus, PL has greater representational capacity.

Serotonergic control of GABAergic inhibition in the lateral amygdala.

Much evidence implicates the serotonergic regulation of the amygdala in anxiety. Thus the present study was undertaken to characterize the influence of serotonin (5-HT) on principal neurons (PNs) of the rat lateral amygdala (LA), using whole cell recordings in vitro. Because inhibition is a major determinant of PN activity, we focused on the control of GABAergic transmission by 5-HT. IPSCs were elicited by local electrical stimulation of LA in the presence of glutamate receptor antagonists. We found that 5-HT reduces GABAA inhibitory postsynaptic currents (IPSCs) via presynaptic 5-HT1B receptors. While the presynaptic inhibition of GABA release also attenuated GABAB currents, this effect was less pronounced than for GABAA currents because 5-HT also induced a competing postsynaptic enhancement of GABAB currents. That is, GABAB currents elicited by pressure application of GABA or baclofen were enhanced by 5-HT. In addition, we obtained evidence suggesting that 5-HT differentially regulates distinct subsets of GABAergic synapses. Indeed, GABAA IPSCs were comprised of two components: a relatively 5-HT-insensitive IPSC that had a fast time course and a 5-HT-sensitive component that had a slower time course. Because the relative contribution of these two components varied depending on whether neurons were recorded at proximity versus at a distance from the stimulating electrodes, we speculate that distinct subtypes of local-circuit cells contribute the two contingents of GABAergic synapses. Overall, our results indicate that 5-HT is a potent regulator of synaptic inhibition in LA.NEW & NOTEWORTHY We report that 5-HT, acting via presynaptic 5-HT1B receptors, attenuates GABAA IPSCs by reducing GABA release in the lateral amygdala (LA). In parallel, 5-HT enhances GABAB currents postsynaptically, such that GABAB inhibitory postsynaptic currents (IPSCs) are relatively preserved from the presynaptic inhibition of GABA release. We also found that the time course of 5-HT-sensitive and -insensitive GABAA IPSCs differ. Together, these results indicate that 5-HT is a potent regulator of synaptic inhibition in LA.

Closed-loop control of gamma oscillations in the amygdala demonstrates their role in spatial memory consolidation.

Gamma is a ubiquitous brain rhythm hypothesized to support cognitive, perceptual, and mnemonic functions by coordinating neuronal interactions. While much correlational evidence supports this hypothesis, direct experimental tests have been lacking. Since gamma occurs as brief bursts of varying frequencies and durations, most existing approaches to manipulate gamma are either too slow, delivered irrespective of the rhythm's presence, not spectrally specific, or unsuitable for bidirectional modulation. Here, we overcome these limitations with an approach that accurately detects and modulates endogenous gamma oscillations, using closed-loop signal processing and optogenetic stimulation. We first show that the rat basolateral amygdala (BLA) exhibits prominent gamma oscillations during the consolidation of contextual memories. We then boost or diminish gamma during consolidation, in turn enhancing or impairing subsequent memory strength. Overall, our study establishes the role of gamma oscillations in memory consolidation and introduces a versatile method for studying fast network rhythms in vivo.

Embracing Complexity in Defensive Networks.

The neural basis of defensive behaviors continues to attract much interest, not only because they are important for survival but also because their dysregulation may be at the origin of anxiety disorders. Recently, a dominant approach in the field has been the optogenetic manipulation of specific circuits or cell types within these circuits to dissect their role in different defensive behaviors. While the usefulness of optogenetics is unquestionable, we argue that this method, as currently applied, fosters an atomistic conceptualization of defensive behaviors, which hinders progress in understanding the integrated responses of nervous systems to threats. Instead, we advocate for a holistic approach to the problem, including observational study of natural behaviors and their neuronal correlates at multiple sites, coupled to the use of optogenetics, not to globally turn on or off neurons of interest, but to manipulate specific activity patterns hypothesized to regulate defensive behaviors.

Basolateral amygdala neurons are activated during threat expectation.

Fear conditioning studies have led to the view that the amygdala contains neurons that signal threat and in turn elicit defensive behaviors through their brain stem and hypothalamic targets. In agreement with this model, a prior unit-recording study in rats performing a seminaturalistic foraging task revealed that many lateral amygdala (LA) neurons are predator responsive. In contrast, our previous study emphasized that most basolateral (BL) amygdala neurons are inhibited at proximity of the predator. However, the two studies used different methods to analyze unit activity, complicating comparisons between them. By applying the same method to the sample of BL neurons we recorded previously, the present study revealed that most principal cells are inhibited by the predator and only 4.5% are activated. Moreover, two-thirds of these cells were also activated by nonthreatening stimuli. In fact, fitting unit activity with a generalized linear model revealed that the only task variables associated with a prevalent positive modulation of BL activity were expectation of the predator's presence and whether the prior trial had been a failure or success. At odds with the threat-coding model of the amygdala, actual confrontation with the predator was usually associated with a widespread inhibition of principal BL neurons. NEW & NOTEWORTHY The basolateral amygdala (BL) is thought to contain neurons that signal threat, in turn eliciting defensive behaviors. In contrast, the present study reports that very few principal BL cells are responsive to threats and that most of them are also activated by nonthreatening stimuli. Yet, expectation of the threat's presence was associated with a prevalent positive modulation of BL activity; actual confrontation with the threat was associated with a widespread inhibition.

Gamma Oscillations in the Basolateral Amygdala: Biophysical Mechanisms and Computational Consequences.

The basolateral nucleus of the amygdala (BL) is thought to support numerous emotional behaviors through specific microcircuits. These are often thought to be comprised of feedforward networks of principal cells (PNs) and interneurons. Neither well-understood nor often considered are recurrent and feedback connections, which likely engender oscillatory dynamics within BL. Indeed, oscillations in the gamma frequency range (40 - 100 Hz) are known to occur in the BL, and yet their origin and effect on local circuits remains unknown. To address this, we constructed a biophysically and anatomically detailed model of the rat BL and its local field potential (LFP) based on the physiological and anatomical literature, along with in vivo and in vitro data we collected on the activities of neurons within the rat BL. Remarkably, the model produced intermittent gamma oscillations (∼50 - 70 Hz) whose properties matched those recorded in vivo, including their entrainment of spiking. BL gamma-band oscillations were generated by the intrinsic circuitry, depending upon reciprocal interactions between PNs and fast-spiking interneurons (FSIs), while connections within these cell types affected the rhythm's frequency. The model allowed us to conduct experimentally impossible tests to characterize the synaptic and spatial properties of gamma. The entrainment of individual neurons to gamma depended on the number of afferent connections they received, and gamma bursts were spatially restricted in the BL. Importantly, the gamma rhythm synchronized PNs and mediated competition between ensembles. Together, these results indicate that the recurrent connectivity of BL expands its computational and communication repertoire.

Detection of Multiway Gamma Coordination Reveals How Frequency Mixing Shapes Neural Dynamics.

A principle of communication technology, frequency mixing, describes how novel oscillations are generated when rhythmic inputs converge on a nonlinearly activating target. As expected given that neurons are nonlinear integrators, it was demonstrated that neuronal networks exhibit mixing in response to imposed oscillations of known frequencies. However, determining when mixing occurs in spontaneous conditions, where weaker or more variable rhythms prevail, has remained impractical. Here, we show that, by exploiting the predicted phase (rather than frequency) relationships between oscillations, the contributions of mixing can be readily identified, even in small samples of noisy data. Assessment of extracellular data using this approach revealed that frequency mixing is widely expressed in a state- and region-dependent manner and that oscillations emerging from mixing entrain unit activity. Frequency mixing is thus intrinsic to the structure of neural activity and contributes importantly to neural dynamics.

Classification of Brainwaves Using Convolutional Neural Network.

Classification of brainwaves in recordings is of considerable interest to neuroscience and medical communities. Classification techniques used presently depend on the extraction of low-level features from the recordings, which in turn affects the classification performance. To alleviate this problem, this paper proposes an end-to-end approach using Convolutional Neural Network (CNN) which has been shown to detect complex patterns in a signal by exploiting its spatiotemporal nature. The present study uses time and frequency axes for the classification using synthesized Local Field Potential (LFP) data. The results are analyzed and compared with the FFT technique. In all the results, the CNN outperforms the FFT by a significant margin especially when the noise level is high. This study also sheds light on certain signal characteristics affecting network performance.

Midline thalamic inputs to the amygdala: Ultrastructure and synaptic targets.

One of the main subcortical inputs to the basolateral nucleus of the amygdala (BL) originates from a group of dorsal thalamic nuclei located at or near the midline, mainly from the central medial (CMT), and paraventricular (PVT) nuclei. Although similarities among the responsiveness of BL, CMT, and PVT neurons to emotionally arousing stimuli suggest that these thalamic inputs exert a significant influence over BL activity, little is known about the synaptic relationships that mediate these effects. Thus, the present study used Phaseolus vulgaris-leucoagglutinin (PHAL) anterograde tracing and electron microscopy to shed light on the ultrastructural properties and synaptic targets of CMT and PVT axon terminals in the rat BL. Virtually all PHAL-positive CMT and PVT axon terminals formed asymmetric synapses. Although CMT and PVT axon terminals generally contacted dendritic spines, a substantial number ended on dendritic shafts. To determine whether these dendritic shafts belonged to principal or local-circuit cells, calcium/calmodulin-dependent protein kinase II (CAMKIIα) immunoreactivity was used as a selective marker of principal BL neurons. In most cases, dendritic shafts postsynaptic to PHAL-labeled CMT and PVT terminals were immunopositive for CaMKIIα. Overall, these results suggest that CMT and PVT inputs mostly target principal BL neurons such that when CMT or PVT neurons fire, little feed-forward inhibition counters their excitatory influence over principal cells. These results are consistent with the possibility that CMT and PVT inputs constitute major determinants of BL activity.

Multi-dimensional Coding by Basolateral Amygdala Neurons.

Conditioned appetitive and aversive responses (CRs) are thought to result from the activation of specific subsets of valence-coding basolateral amygdala (BLA) neurons. Under this model, the responses of BLA cells to conditioned stimuli (CSs) and the activity that drives CRs are closely related. We tested the strength of this correlation using a task where rats could emit different CRs in response to the same CSs. At odds with this model, the CS responses and CR-related activity of individual BLA cells were separable. Moreover, while the incidence of valence-coding cells did not exceed chance, at the population level there was similarity between valence coding for CSs and CRs. In fact, both lateral and basolateral neurons concurrently encoded multiple task features and behaviors. Thus, conditioned emotional behaviors may not depend on the recruitment of single cells that explicitly encode individual task variables but from multiplexed representations distributed across the BLA.

Optogenetic Study of Anterior BNST and Basomedial Amygdala Projections to the Ventromedial Hypothalamus.

The basomedial amygdala (BM) influences the ventromedial nucleus of the hypothalamus (VMH) through direct glutamatergic projections as well as indirectly, through the anterior part of the bed nucleus of the stria terminalis (BNSTa). However, BM and BNSTa axons end in a segregated fashion in VMH. BM projects to the core of VMH, where VMH's projection cells are located, whereas BNSTa projects to the shell of VMH, where GABAergic cells that inhibit core neurons are concentrated. However, the consequences of this dual regulation of VMH by BM and BNSTa are unknown. To study this question, we recorded the responses of VMH's shell and core neurons to the optogenetic activation of BM or BNSTa inputs in transgenic mice that selectively express Cre-recombinase in glutamatergic or GABAergic neurons. Glutamatergic BM inputs fired most core neurons but elicited no response in GABAergic shell neurons. Following BM infusions of AAV-EF1α-DIO-hChR2-mCherry in Vgat-ires-Cre-Ai6 mice, no anterograde labeling was observed in the VMH, suggesting that GABAergic BM neurons do not project to the VMH. In contrast, BNSTa sent mostly GABAergic projections that inhibited both shell and core neurons. However, BNSTa-evoked IPSPs had a higher amplitude in shell neurons. Since we also found that activation of GABAergic shell neurons causes an inhibition of core neurons, these results suggest that depending on the firing rate of shell neurons, BNSTa inputs could elicit a net inhibition or disinhibition of core neurons. Thus, the dual regulation of VMH by BM and BNSTa imparts flexibility to this regulator of defensive and social behaviors.

Glutamatergic and gabaergic ventral BNST neurons differ in their physiological properties and responsiveness to noradrenaline.

The bed nucleus of the stria terminalis (BNST) regulates defensive responses to threats and its anteroventral portion (BNST-AV) is involved. BNST-AV contains a minority of glutamatergic neurons scattered among a dominant population of GABAergic cells. There is evidence that these two cell types might exert opposite influences, the former promoting and the latter reducing anxiety. Although GABAergic cells greatly outnumber glutamatergic neurons in BNST-AV, in some circumstances the influence of glutamatergic cells appears to predominate. Related to this, BNST-AV receives a very strong noradrenaline (NA) input and negative emotional states are associated with a marked rise of NA concentration in BNST-AV. However, it is currently unclear whether NA differentially alters the excitability of glutamatergic and GABAergic BNST-AV neurons. Thus, to shed light on how BNST-AV regulates negative emotional states, the present study compared the physiological properties and NA responsiveness of glutamatergic and GABAergic BNST-AV neurons using whole-cell recordings in transgenic mice that express a fluorescent reporter in either cell group. We found that glutamatergic cells had a slightly more complex morphology than the GABAergic cells, a higher intrinsic excitability, and a different responsiveness to NA. Indeed, while NA inhibited EPSPs in both cell types through α1 and α2 adrenoreceptors, the EPSP reduction seen in glutamatergic cells had a lower amplitude and a shorter duration than in GABAergic cells. These differences were due to the presence of a ß-receptor-mediated EPSP enhancement in the glutamatergic cells. Together, our results suggest that multiple properties contribute to the disproportionate influence of glutamatergic BNST-AV neurons.

Vigilance-Associated Gamma Oscillations Coordinate the Ensemble Activity of Basolateral Amygdala Neurons.

Principal basolateral amygdala (BL) neurons profoundly influence motivated behaviors, yet few of them are activated by emotionally valenced stimuli. Here, we show that a likely explanation for this paradox is the synchronizing influence of the high-gamma rhythm. High-gamma (75-95 Hz) entrained BL firing more strongly than all other rhythms. It was most pronounced during states of increased vigilance, when rats were apprehensive. Relative to behavioral states, high-gamma produced minor changes in firing rates yet dramatic increases in synchrony. Moreover, connected pairs of cells showed similarly high levels of entrainment and synchronization. Unexpectedly, prefrontal- and accumbens-projecting cells, respectively, showed high and low entrainment by high-gamma, indicating that this rhythm differentially synchronizes the activity of BL neurons projecting to specific sites. Overall, our findings suggest that individual BL neurons encode information not only by changing their firing rates, but also by synchronizing their collective activity, amplifying their impact on target structures.