Eszopiclone and Zolpidem Produce Opposite Effects on Hippocampal Ripple Density

Clinical populations have memory deficits linked to sleep oscillations that can potentially be treated with sleep medications. Eszopiclone and zolpidem (two non-benzodiazepine hypnotics) both enhance sleep spindles. Zolpidem improved sleep-dependent memory consolidation in humans, but eszopiclone did not. These divergent results may reflect that the two drugs have different effects on hippocampal ripple oscillations, which correspond to the reactivation of neuronal ensembles that represent previous waking activity and contribute to memory consolidation. We used extracellular recordings in the CA1 region of rats and systemic dosing of eszopiclone and zolpidem to test the hypothesis that these two drugs differentially affect hippocampal ripples and spike activity. We report evidence that eszopiclone makes ripples sparser, while zolpidem increases ripple density. In addition, eszopiclone led to a drastic decrease in spike firing, both in putative pyramidal cells and interneurons, while zolpidem did not substantially alter spiking. These results provide an explanation of the different effects of eszopiclone and zolpidem on memory in human studies and suggest that sleep medications can be used to regulate hippocampal ripple oscillations, which are causally linked to sleep-dependent memory consolidation.

A luciferase prosubstrate and a red bioluminescent calcium indicator to image neuronal activity

Genetically encoded fluorescent indicators have been broadly used to monitor neuronal activity in live animals, but invasive surgical procedures are required. This study presents a functional bioluminescence imaging (fBLI) method for recording the activity of neuronal ensembles in the brain in awake mice. We developed a luciferase prosubstrate activatable in vivo by nonspecific esterase to enhance the brain delivery of the luciferin. We further engineered a bright, bioluminescent indicator with robust responsiveness to calcium ions (Ca2+) and appreciable emission above 600 nm. Integration of these advantageous components enabled the imaging of Ca2+ dynamics in awake mice minimally invasively with excellent signal- to-background and subsecond temporal resolution. This study thus establishes a new paradigm for studying brain functions in health and disease.

Lost in time: rethinking duration estimation outside the brain.

Our perception of the passage of time can suffer from significant distortions as time flies when we are busy and drags when we are bored. A prominent mechanistic model proposes that this perceptual volatility reflects changes in the activity dynamics of distributed neuronal ensembles referred to as population clocks because they encode time. In this framework, time is understood similarly to space (both can be segmented in seconds or centimeters) and duration estimation is primarily internal (the brain tells time). Here, I challenge this framework from the angle of Bergson’s proposal that the inner experience of time is unlike space because it is ever-changing and indivisible (2 successive seconds are not experienced equivalently). Quantifying and communicating this inner experience requires its externalization and immobilization through distance measurements derived from stereotyped movements and spatial metaphors (“short/long” durations; time “flies/drags”), which explains the habit of thinking time like space. In support of Bergson’s proposal, humans and animals heavily rely on movements in a variety of duration estimation tasks and the neural underpinnings of duration estimates overlap those of motor control and spatial navigation. Thus, philosophical and empirical arguments question whether duration estimation is fundamentally internal. Rather than being explained by ad hoc changes in the speed of population clocks, the puzzle of the volatility of time perception might resolve itself by considering that living beings lack the ability to internally measure time, which they compensate through interactions with regularities afforded by the world and symbolic representation drawn from space.

Hippocampal-Prefrontal cortex network dynamics predict performance during retrieval in a context-guided object memory task

Remembering life episodes is a complex process that requires the interaction between multiple brain areas. It is thought that contextual information provided by the hippocampus (HPC) can trigger the recall of a past event through the activation of medial prefrontal cortex (mPFC) neuronal ensembles, but the underlying mechanisms remain poorly understood. Indeed, little is known about how the vHPC and mPFC are coordinated during a contextual-guided recall of an object recognition memory. To address this, we performed electrophysiological recordings in behaving rats during the retrieval phase of the object-in-context memory task (OIC). Coherence, phase locking and theta amplitude correlation analysis showed an increase in vHPC-mPFC LFP synchronization in the theta range when animals explore contextually mismatched objects. Moreover, we identified ensembles of putative pyramidal cells in the mPFC that encode specific object-context associations. Interestingly, the increase of vHPC-mPFC synchronization during exploration of the contextually mismatched object and the preference of mPFC incongruent object neurons predicts the animal's performance during the resolution of the OIC task. Altogether, these results identify changes in vHPC-mPFC synchronization and mPFC ensembles encoding specific object-context associations likely involved in the recall of past events.

Periaqueductal gray neurons encode the sequential motor program in hunting behavior of mice

Sequential encoding of motor programs is essential for behavior generation. However, whether it is critical for instinctive behavior is still largely unknown. Mouse hunting behavior typically contains a sequential motor program, including the prey search, chase, attack, and consumption. Here, we reveal that the neuronal activity in the lateral periaqueductal gray (LPAG) follows a sequential pattern and is time-locked to different hunting actions. Optrode recordings and photoinhibition demonstrate that LPAGVgat neurons are required for the prey detection, chase and attack, while LPAGVglut2 neurons are selectively required for the attack. Ablation of inputs that could trigger hunting, including the central amygdala, the lateral hypothalamus, and the zona incerta, interrupts the activity sequence pattern and substantially impairs hunting actions. Therefore, our findings reveal that periaqueductal gray neuronal ensembles encode the sequential hunting motor program, which might provide a framework for decoding complex instinctive behaviors.

Co-allocation to overlapping dendritic branches in the retrosplenial cortex integrates contextual memories across time

Events occurring close in time are often linked in memory, providing an episodic timeline and a framework for those memories. Recent studies suggest that memories acquired close in time are encoded by overlapping neuronal ensembles, and that this overlap is necessary for memory linking. Transient increases in neuronal excitability drive this ensemble overlap, but whether dendritic plasticity plays a role in linking memories is unknown. Here, we show that contextual memory linking is not only dependent on ensemble overlap in the retrosplenial cortex (RSC), but also on RSC branch-specific dendritic allocation mechanisms. Using longitudinal two-photon calcium imaging of RSC dendrites, we show that the same dendritic segments are preferentially activated by two linked (but not independent) contextual memories, and that spine clusters added after each of two linked (but not independent) contextual memories are allocated to the same dendritic segments. Importantly, with a novel optogenetic tool, selectively targeted to activated dendritic segments following learning, we show that reactivation of dendrites tagged during the first context exploration is sufficient to link two contextual memories. These results demonstrate a causal role for dendritic mechanisms in memory linking and reveal a novel set of rules that govern how linked, and independent memories are allocated to dendritic compartments.

Dual polarity voltage imaging reveals subthreshold dynamics and concurrent spiking patterns of multiple neuron-types

Genetically encoded fluorescent voltage indicators are ideally suited to reveal the millisecond-scale interactions among and between distinct, targeted cell populations. However, current indicator families lack the requisite sensitivity for in vivo multipopulation imaging. We describe high-performance green and red sensors, Ace-mNeon2 and VARNAM2, and their reverse response-polarity variants, pAce and pAceR. Our indicators enable 0.4-1 kHz voltage recordings from >50 neurons per field-of-view in awake mice and ~30-min continuous imaging in flies. Using dual-polarity multiplexed imaging, we uncovered behavioral state-dependent interactions between distinct neocortical subclasses, as well as contributions to hippocampal field potentials from non-overlapping projection neuronal ensembles. By combining three mutually compatible indicators, we demonstrate concurrent triple-population voltage imaging. Our approach will empower investigations of the dynamic interplay between neuronal subclasses at single-spike resolution.

CRHCeA→VTA inputs inhibit the positive ensembles to induce negative effect of opiate withdrawal

Plasticity of neurons in the ventral tegmental area (VTA) is critical for establishment of drug dependence. However, the remodeling of the circuits mediating the transition between positive and negative effect remains unclear. Here, we used neuronal activity-dependent labeling technique to characterize and temporarily control the VTA neuronal ensembles recruited by the initial morphine exposure (morphine-positive ensembles, Mor-Ens). Mor-Ens preferentially projected to NAc, and induced dopamine-dependent positive reinforcement. Electrophysiology and rabies viral tracing revealed the preferential connections between the VTA-projective corticotrophin-releasing hormone (CRH) neurons of central amygdala (CRHCeA→VTA) and Mor-Ens, which was enhanced after escalating morphine exposure and mediated the negative effect during opiate withdrawal. Pharmacologic intervention or CRISPR-mediated repression of CRHR1 in Mor-Ens weakened the inhibitory CRHCeA→VTA inputs, and alleviated the negative effect during opiate withdrawal. These data suggest that neurons encoding opioid reward experience are inhibited by enhanced CRHCeA→VTA inputs induced by chronic morphine exposure, leading to negative effect during opiate withdrawal, and provide new insight into the pathological changes in VTA plasticity after drug abuse and mechanism of opiate dependence.

Prefrontal neural ensembles encode an internal model of visual sequences and their violations

Theories of predictive coding hypothesize that cortical networks learn internal models of environmental regularities to generate expectations that are constantly compared with sensory inputs. The prefrontal cortex (PFC) is thought to be critical for predictive coding. Here, we show how prefrontal neuronal ensembles encode a detailed internal model of sequences of visual events and their violations. We recorded PFC ensembles in a visual local-global sequence paradigm probing low and higher-order predictions and mismatches. PFC ensembles formed distributed, overlapping representations for all aspects of the dynamically unfolding sequences, including information about image identity as well as abstract information about ordinal position, anticipated sequence pattern, mismatches to local and global structure, and model updates. Model and mismatch signals were mixed in the same ensembles, suggesting a revision of predictive processing models that consider segregated processing. We conclude that overlapping prefrontal ensembles may collectively encode all aspects of an ongoing visual experience, including anticipation, perception, and surprise.