Fear and safety engage competing patterns of theta-gamma coupling in the basolateral amygdala.

Theta oscillations synchronize the basolateral amygdala (BLA) with the hippocampus (HPC) and medial prefrontal cortex (mPFC) during fear expression. The role of gamma-frequency oscillations in the BLA is less well characterized. We examined gamma- and theta-frequency activity in recordings of neural activity from the BLA-HPC-mPFC circuit during fear conditioning, extinction, and exposure to an open field. In the BLA, slow (40-70 Hz) and fast (70-120 Hz) gamma oscillations were coupled to distinct phases of the theta cycle and reflected synchronous high-frequency unit activity. During periods of fear, BLA theta-fast gamma coupling was enhanced, while fast gamma power was suppressed. Periods of relative safety were associated with enhanced BLA fast gamma power, mPFC-to-BLA directionality, and strong coupling of BLA gamma to mPFC theta. These findings suggest that switches between states of fear and safety are mediated by changes in BLA gamma coupling to competitive theta frequency inputs.

Prefrontal entrainment of amygdala activity signals safety in learned fear and innate anxiety.

Successfully differentiating safety from danger is an essential skill for survival. While decreased activity in the medial prefrontal cortex (mPFC) is associated with fear generalization in animals and humans, the circuit-level mechanisms used by the mPFC to discern safety are not clear. To answer this question, we recorded activity in the mPFC, basolateral amygdala (BLA) and dorsal and ventral hippocampus in mice during exposure to learned (differential fear conditioning) and innate (open field) anxiety. We found increased synchrony between the mPFC and BLA in the theta frequency range (4-12 Hz) only in animals that differentiated between averseness and safety. Moreover, during recognized safety across learned and innate protocols, BLA firing became entrained to theta input from the mPFC. These data suggest that selective tuning of BLA firing to mPFC input provides a safety-signaling mechanism whereby the mPFC taps into the microcircuitry of the amygdala to diminish fear.

Single units in the medial prefrontal cortex with anxiety-related firing patterns are preferentially influenced by ventral hippocampal activity.

The medial prefrontal cortex (mPFC) and ventral hippocampus (vHPC) functionally interact during innate anxiety tasks. To explore the consequences of this interaction, we examined task-related firing of single units from the mPFC of mice exploring standard and modified versions of the elevated plus maze (EPM), an innate anxiety paradigm. Hippocampal local field potentials (LFPs) were simultaneously monitored. The population of mPFC units distinguished between safe and aversive locations within the maze, regardless of the nature of the anxiogenic stimulus. Strikingly, mPFC units with stronger task-related activity were more strongly coupled to theta-frequency activity in the vHPC LFP. Lastly, task-related activity was inversely correlated with behavioral measures of anxiety. These results clarify the role of the vHPC-mPFC circuit in innate anxiety and underscore how specific inputs may be involved in the generation of behaviorally relevant neural activity within the mPFC.

Cross-correlation of instantaneous amplitudes of field potential oscillations: a straightforward method to estimate the directionality and lag between brain areas.

Researchers performing multi-site recordings are often interested in identifying the directionality of functional connectivity and estimating lags between sites. Current techniques for determining directionality require spike trains or involve multivariate autoregressive modeling. However, it is often difficult to sample large numbers of spikes from multiple areas simultaneously, and modeling can be sensitive to noise. A simple, model-independent method to estimate directionality and lag using local field potentials (LFPs) would be of general interest. Here we describe such a method using the cross-correlation of the instantaneous amplitudes of filtered LFPs. The method involves four steps. First, LFPs are band-pass filtered; second, the instantaneous amplitude of the filtered signals is calculated; third, these amplitudes are cross-correlated and the lag at which the cross-correlation peak occurs is determined; fourth, the distribution of lags obtained is tested to determine if it differs from zero. This method was applied to LFPs recorded from the ventral hippocampus and the medial prefrontal cortex in awake behaving mice. The results demonstrate that the hippocampus leads the mPFC, in good agreement with the time lag calculated from the phase locking of mPFC spikes to vHPC LFP oscillations in the same dataset. We also compare the amplitude cross-correlation method to partial directed coherence, a commonly used multivariate autoregressive model-dependent method, and find that the former is more robust to the effects of noise. These data suggest that the cross-correlation of instantaneous amplitude of filtered LFPs is a valid method to study the direction of flow of information across brain areas.

Synchronized activity between the ventral hippocampus and the medial prefrontal cortex during anxiety.

The ventral hippocampus, unlike its dorsal counterpart, is required for anxiety-like behavior. The means by which it acts are unknown. We hypothesized that the hippocampus synchronizes with downstream targets that influence anxiety, such as the medial prefrontal cortex (mPFC). To test this hypothesis, we recorded mPFC and hippocampal activity in mice exposed to two anxiogenic arenas. Theta-frequency activity in the mPFC and ventral, but not dorsal, hippocampus was highly correlated at baseline, and this correlation increased in both anxiogenic environments. Increases in mPFC theta power predicted avoidance of the aversive compartments of each arena and were larger in serotonin 1A receptor knockout mice, a genetic model of increased anxiety-like behavior. These results suggest a role for theta-frequency synchronization between the ventral hippocampus and the mPFC in anxiety. They are consistent with the notion that such synchronization is a general mechanism by which the hippocampus communicates with downstream structures of behavioral relevance.