RESUMEN
Memory encoding and retrieval rely on specific interactions across multiple brain areas. Although connections between individual brain areas have been extensively studied, the anatomical and functional specificity of neuronal circuit organization underlying information transfer across multiple brain areas remains unclear. Here, we combine transsynaptic viral tracing, optogenetic manipulations, and calcium dynamics recordings to dissect the multisynaptic functional connectivity of the amygdala. We identify a distinct basolateral amygdala (BLA) subpopulation that connects disynaptically to the periaqueductal gray (PAG) via the central amygdala (CeA). This disynaptic pathway serves as a core circuit element necessary for the learning and expression of conditioned fear and exhibits learning-related plasticity. Together, our findings demonstrate the utility of multisynaptic approaches for functional circuit analysis and indicate that disynaptic specificity may be a general feature of neuronal circuit organization.
RESUMEN
Coping with threatening situations requires both identifying stimuli that predict danger and selecting adaptive behavioural responses to survive1. The dorsomedial prefrontal cortex (dmPFC) is a critical structure that is involved in the regulation of threat-related behaviour2-4. However, it is unclear how threat-predicting stimuli and defensive behaviours are associated within prefrontal networks to successfully drive adaptive responses. Here we used a combination of extracellular recordings, neuronal decoding approaches, pharmacological and optogenetic manipulations to show that, in mice, threat representations and the initiation of avoidance behaviour are dynamically encoded in the overall population activity of dmPFC neurons. Our data indicate that although dmPFC population activity at stimulus onset encodes sustained threat representations driven by the amygdala, it does not predict action outcome. By contrast, transient dmPFC population activity before the initiation of action reliably predicts avoided from non-avoided trials. Accordingly, optogenetic inhibition of prefrontal activity constrained the selection of adaptive defensive responses in a time-dependent manner. These results reveal that the adaptive selection of defensive responses relies on a dynamic process of information linking threats with defensive actions, unfolding within prefrontal networks.
Asunto(s)
Reacción de Prevención , Mecanismos de Defensa , Neuronas/fisiología , Corteza Prefrontal/fisiología , Amígdala del Cerebelo/fisiología , Animales , Miedo , Masculino , Ratones , Ratones Endogámicos C57BL , OptogenéticaRESUMEN
Learning drives behavioral adaptations necessary for survival. While plasticity of excitatory projection neurons during associative learning has been extensively studied, little is known about the contributions of local interneurons. Using fear conditioning as a model for associative learning, we found that behaviorally relevant, salient stimuli cause learning by tapping into a local microcircuit consisting of precisely connected subtypes of inhibitory interneurons. By employing deep-brain calcium imaging and optogenetics, we demonstrate that vasoactive intestinal peptide (VIP)-expressing interneurons in the basolateral amygdala are activated by aversive events and provide a mandatory disinhibitory signal for associative learning. Notably, VIP interneuron responses during learning are strongly modulated by expectations. Our findings indicate that VIP interneurons are a central component of a dynamic circuit motif that mediates adaptive disinhibitory gating to specifically learn about unexpected, salient events, thereby ensuring appropriate behavioral adaptations.
Asunto(s)
Aprendizaje por Asociación/fisiología , Interneuronas/fisiología , Inhibición Neural/fisiología , Filtrado Sensorial/fisiología , Péptido Intestinal Vasoactivo/fisiología , Amígdala del Cerebelo/fisiología , Animales , Condicionamiento Psicológico/fisiología , Miedo/psicología , Femenino , Masculino , Ratones , Ratones Transgénicos , OptogenéticaRESUMEN
When faced with threat, the survival of an organism is contingent upon the selection of appropriate active or passive behavioural responses. Freezing is an evolutionarily conserved passive fear response that has been used extensively to study the neuronal mechanisms of fear and fear conditioning in rodents. However, rodents also exhibit active responses such as flight under natural conditions. The central amygdala (CEA) is a forebrain structure vital for the acquisition and expression of conditioned fear responses, and the role of specific neuronal sub-populations of the CEA in freezing behaviour is well-established. Whether the CEA is also involved in flight behaviour, and how neuronal circuits for active and passive fear behaviour interact within the CEA, are not yet understood. Here, using in vivo optogenetics and extracellular recordings of identified cell types in a behavioural model in which mice switch between conditioned freezing and flight, we show that active and passive fear responses are mediated by distinct and mutually inhibitory CEA neurons. Cells expressing corticotropin-releasing factor (CRF+) mediate conditioned flight, and activation of somatostatin-positive (SOM+) neurons initiates passive freezing behaviour. Moreover, we find that the balance between conditioned flight and freezing behaviour is regulated by means of local inhibitory connections between CRF+ and SOM+ neurons, indicating that the selection of appropriate behavioural responses to threat is based on competitive interactions between two defined populations of inhibitory neurons, a circuit motif allowing for rapid and flexible action selection.
Asunto(s)
Reacción de Fuga/fisiología , Miedo/fisiología , Miedo/psicología , Reacción Cataléptica de Congelación/fisiología , Inhibición Neural , Neuronas/fisiología , Animales , Núcleo Amigdalino Central/citología , Núcleo Amigdalino Central/fisiología , Hormona Liberadora de Corticotropina/metabolismo , Locomoción/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Neurológicos , Vías Nerviosas , Optogenética , Somatostatina/metabolismoRESUMEN
Precise spike timing through the coordination and synchronization of neuronal assemblies is an efficient and flexible coding mechanism for sensory and cognitive processing. In cortical and subcortical areas, the formation of cell assemblies critically depends on neuronal oscillations, which can precisely control the timing of spiking activity. Whereas this form of coding has been described for sensory processing and spatial learning, its role in encoding emotional behaviour remains unknown. Fear behaviour relies on the activation of distributed structures, among which the dorsal medial prefrontal cortex (dmPFC) is known to be critical for fear memory expression. In the dmPFC, the phasic activation of neurons to threat-predicting cues, a spike-rate coding mechanism, correlates with conditioned fear responses and supports the discrimination between aversive and neutral stimuli. However, this mechanism does not account for freezing observed outside stimuli presentations, and the contribution of a general spike-time coding mechanism for freezing in the dmPFC remains to be established. Here we use a combination of single-unit and local field potential recordings along with optogenetic manipulations to show that, in the dmPFC, expression of conditioned fear is causally related to the organization of neurons into functional assemblies. During fear behaviour, the development of 4 Hz oscillations coincides with the activation of assemblies nested in the ascending phase of the oscillation. The selective optogenetic inhibition of dmPFC neurons during the ascending or descending phases of this oscillation blocks and promotes conditioned fear responses, respectively. These results identify a novel phase-specific coding mechanism, which dynamically regulates the development of dmPFC assemblies to control the precise timing of fear responses.
Asunto(s)
Miedo/fisiología , Vías Nerviosas , Corteza Prefrontal/citología , Corteza Prefrontal/fisiología , Animales , Condicionamiento Clásico , Reacción Cataléptica de Congelación , Masculino , Memoria/fisiología , Ratones , Ratones Endogámicos C57BL , Neuronas/fisiología , Optogenética , Factores de TiempoRESUMEN
Fear expression relies on the coordinated activity of prefrontal and amygdala circuits, yet the mechanisms allowing long-range network synchronization during fear remain unknown. Using a combination of extracellular recordings, pharmacological and optogenetic manipulations, we found that freezing, a behavioral expression of fear, temporally coincided with the development of sustained, internally generated 4-Hz oscillations in prefrontal-amygdala circuits. 4-Hz oscillations predict freezing onset and offset and synchronize prefrontal-amygdala circuits. Optogenetic induction of prefrontal 4-Hz oscillations coordinates prefrontal-amygdala activity and elicits fear behavior. These results unravel a sustained oscillatory mechanism mediating prefrontal-amygdala coupling during fear behavior.
Asunto(s)
Amígdala del Cerebelo/fisiología , Relojes Biológicos/fisiología , Miedo/fisiología , Miedo/psicología , Optogenética/métodos , Corteza Prefrontal/fisiología , Estimulación Acústica/efectos adversos , Animales , Condicionamiento Psicológico/fisiología , Extinción Psicológica/fisiología , Masculino , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Vías Nerviosas/fisiologíaRESUMEN
Recent technological developments, such as single unit recordings coupled to optogenetic approaches, have provided unprecedented knowledge about the precise neuronal circuits contributing to the expression and recovery of conditioned fear behavior. These data have provided an understanding of the contributions of distinct brain regions such as the amygdala, prefrontal cortex, hippocampus, and periaqueductal gray matter to the control of conditioned fear behavior. Notably, the precise manipulation and identification of specific cell types by optogenetic techniques have provided novel avenues to establish causal links between changes in neuronal activity that develop in dedicated neuronal structures and the short and long-lasting expression of conditioned fear memories. In this review, we provide an update on the key neuronal circuits and cell types mediating conditioned fear expression and recovery and how these new discoveries might refine therapeutic approaches for psychiatric conditions such as anxiety disorders and posttraumatic stress disorder.
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Encéfalo/patología , Encéfalo/fisiología , Miedo , Vías Nerviosas/fisiología , Recuperación de la Función/fisiología , Animales , Ansiedad/patología , Ansiedad/terapia , Humanos , OptogenéticaAsunto(s)
Conducta Animal/fisiología , Miedo/fisiología , Neuronas GABAérgicas/fisiología , Interneuronas/fisiología , Proteínas del Tejido Nervioso/análisis , Parvalbúminas/análisis , Corteza Prefrontal/fisiología , Animales , Trastornos de Ansiedad/fisiopatología , Reacción de Prevención/fisiología , Mapeo Encefálico , Emociones/fisiología , Reacción Cataléptica de Congelación/fisiología , Neuronas GABAérgicas/química , Humanos , Interneuronas/química , Sistema Límbico/fisiología , Red Nerviosa/fisiología , Corteza Prefrontal/citología , Transmisión Sináptica/fisiología , Ritmo TetaRESUMEN
High-frequency intake and high drug-induced seeking are associated with cocaine addiction in both human and animals. However, their relationships and neurobiological underpinnings remain hypothetical. The medial prefrontal cortex (mPFC), basolateral amygdala (BLA), and nucleus accumbens (NAc) have been shown to have a role in cocaine seeking. However, their involvement in regulating high-frequency intake and high cocaine-induced seeking is unclear. We manipulated frequency of cocaine self-administration and investigated whether it influenced cocaine seeking. The contribution of the aforementioned structures was evaluated using changes in expression of the immediate early gene c-Fos and targeted optogenetic manipulations. Rats that self-administered at High frequency (short inter-infusion intervals allowed by short time-out) showed higher cocaine-induced seeking than low frequency rats (long inter-infusions intervals imposed by long time-out), as measured with cocaine-induced reinstatement. c-Fos was enhanced in High frequency rats in the prelimbic (PL) and infralimbic (IL) areas of the mPFC, the BLA, and the NAc core and shell. Correlational analysis of c-Fos revealed that the PL was a critical node strongly correlated with both the IL and NAc core in High frequency rats. Targeted optogenetic inactivation of the PL decreased cocaine-induced reinstatement, but increased cocaine self-administration, in High frequency rats. In contrast, optogenetic activation of the PL had no effect on Low frequency rats. Thus, high-frequency intake promotes a PL-dependent control of cocaine seeking, with the PL exerting a facilitatory or inhibitory effect, depending on operant contingencies. Individual differences in cocaine-induced PL activation might be a source of vulnerability for poorly controlled cocaine-induced seeking and/or cocaine intake.
Asunto(s)
Encéfalo/efectos de los fármacos , Trastornos Relacionados con Cocaína/fisiopatología , Cocaína/administración & dosificación , Inhibidores de Captación de Dopamina/administración & dosificación , Comportamiento de Búsqueda de Drogas/efectos de los fármacos , Animales , Encéfalo/fisiopatología , Comportamiento de Búsqueda de Drogas/fisiología , Extinción Psicológica/efectos de los fármacos , Extinción Psicológica/fisiología , Masculino , Neuronas/efectos de los fármacos , Neuronas/fisiología , Optogenética , Proteínas Proto-Oncogénicas c-fos/metabolismo , Ratas Sprague-Dawley , Autoadministración , Factores de TiempoRESUMEN
Synchronization of spiking activity in neuronal networks is a fundamental process that enables the precise transmission of information to drive behavioural responses. In cortical areas, synchronization of principal-neuron spiking activity is an effective mechanism for information coding that is regulated by GABA (γ-aminobutyric acid)-ergic interneurons through the generation of neuronal oscillations. Although neuronal synchrony has been demonstrated to be crucial for sensory, motor and cognitive processing, it has not been investigated at the level of defined circuits involved in the control of emotional behaviour. Converging evidence indicates that fear behaviour is regulated by the dorsomedial prefrontal cortex (dmPFC). This control over fear behaviour relies on the activation of specific prefrontal projections to the basolateral complex of the amygdala (BLA), a structure that encodes associative fear memories. However, it remains to be established how the precise temporal control of fear behaviour is achieved at the level of prefrontal circuits. Here we use single-unit recordings and optogenetic manipulations in behaving mice to show that fear expression is causally related to the phasic inhibition of prefrontal parvalbumin interneurons (PVINs). Inhibition of PVIN activity disinhibits prefrontal projection neurons and synchronizes their firing by resetting local theta oscillations, leading to fear expression. Our results identify two complementary neuronal mechanisms mediated by PVINs that precisely coordinate and enhance the neuronal activity of prefrontal projection neurons to drive fear expression.
Asunto(s)
Miedo/fisiología , Interneuronas/metabolismo , Inhibición Neural/fisiología , Parvalbúminas/metabolismo , Corteza Prefrontal/citología , Corteza Prefrontal/fisiología , Potenciales de Acción , Amígdala del Cerebelo/fisiología , Animales , Condicionamiento Psicológico , Extinción Psicológica , Miedo/psicología , Masculino , Memoria/fisiología , Ratones , Ratones Endogámicos C57BL , Modelos Neurológicos , Vías Nerviosas , Optogenética , Ritmo TetaRESUMEN
Learning causes a change in how information is processed by neuronal circuits. Whereas synaptic plasticity, an important cellular mechanism, has been studied in great detail, we know much less about how learning is implemented at the level of neuronal circuits and, in particular, how interactions between distinct types of neurons within local networks contribute to the process of learning. Here we show that acquisition of associative fear memories depends on the recruitment of a disinhibitory microcircuit in the mouse auditory cortex. Fear-conditioning-associated disinhibition in auditory cortex is driven by foot-shock-mediated cholinergic activation of layer 1 interneurons, in turn generating inhibition of layer 2/3 parvalbumin-positive interneurons. Importantly, pharmacological or optogenetic block of pyramidal neuron disinhibition abolishes fear learning. Together, these data demonstrate that stimulus convergence in the auditory cortex is necessary for associative fear learning to complex tones, define the circuit elements mediating this convergence and suggest that layer-1-mediated disinhibition is an important mechanism underlying learning and information processing in neocortical circuits.
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Corteza Auditiva/fisiología , Condicionamiento Clásico/fisiología , Miedo/fisiología , Miedo/psicología , Vías Nerviosas/fisiología , Animales , Corteza Auditiva/citología , Corteza Auditiva/efectos de los fármacos , Condicionamiento Clásico/efectos de los fármacos , Electrochoque , Extremidades/inervación , Extremidades/fisiología , Miedo/efectos de los fármacos , Interneuronas/citología , Interneuronas/efectos de los fármacos , Interneuronas/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Neurológicos , Red Nerviosa/citología , Red Nerviosa/efectos de los fármacos , Red Nerviosa/fisiología , Inhibición Neural/efectos de los fármacos , Inhibición Neural/fisiología , Vías Nerviosas/citología , Vías Nerviosas/efectos de los fármacos , Antagonistas Nicotínicos/farmacología , Células Piramidales/efectos de los fármacos , Células Piramidales/fisiología , Receptores Nicotínicos/metabolismoRESUMEN
Morphine is a highly potent analgesic with high addictive potential in specific contexts. Although dopamine neurons of the ventral tegmental area (VTA) are widely believed to play an essential role in the development of drug addiction, neuronal circuits underlying morphine action on dopamine neurons have not been fully elucidated. Here we combined in vivo electrophysiology, tract-tracing experiments, and targeted neuronal inactivation to dissect a neural circuit for acute morphine action on dopamine neurons in rats. We found that in vivo, morphine targets the GABAergic tail of the VTA, also called the rostromedial tegmental nucleus, to increase the firing of dopamine neurons through the activation of VTA µ opioid receptors expressed on tail of the VTA/rostromedial tegmental nucleus efferents. Our data also reveal that in the absence of VTA glutamatergic tone, there is no morphine-induced activation of dopamine neurons. These results define the anatomical organization and functional role of a neural circuit for acute morphine action on dopamine neurons.