Arnaud Malvache

Internally Recurring Hippocampal Sequences as a Population Template of Spatiotemporal Information

Summary The hippocampus is essential for spatiotemporal cognition. Sequences of neuronal activation provide a substrate for this fundamental function. At the behavioral timescale, these sequences have been shown to occur either in the presence of successive external landmarks or through internal mechanisms within an episodic memory task. In both cases, activity is externally constrained by the organization of the task and by the size of the environment explored. Therefore, it remains unknown whether hippocampal activity can self-organize into a default mode in the absence of any external memory demand or spatiotemporal boundary. Here we show that, in the presence of self-motion cues, a population code integrating distance naturally emerges in the hippocampus in the form of recurring sequences. These internal dynamics clamp spontaneous travel since run distance distributes into integer multiples of the span of these sequences. These sequences may thus guide navigation when external landmarks are reduced.

Awake hippocampal reactivations project onto orthogonal neuronal assemblies

The chained activation of neuronal assemblies is thought to support major cognitive processes, including memory. In the hippocampus, this is observed during population bursts often associated with sharp-wave ripples, in the form of an ordered reactivation of neurons. However, the organization and lifetime of these assemblies remain unknown. We used calcium imaging to map patterns of synchronous neuronal activation in the CA1 region of awake mice during runs on a treadmill. The patterns were composed of the recurring activation of anatomically intermingled, but functionally orthogonal, assemblies. These assemblies reactivated discrete temporal segments of neuronal sequences observed during runs and could be stable across consecutive days. A binding of these assemblies into longer chains revealed temporally ordered replay. These modules may represent the default building blocks for encoding or retrieving experience.

GABAergic inhibition shapes interictal dynamics in awake epileptic mice

Epilepsy is characterized by recurrent seizures and brief, synchronous bursts called interictal spikes that are present in-between seizures and observed as transient events in EEG signals. While GABAergic transmission is known to play an important role in shaping healthy brain activity, the role of inhibition in these pathological epileptic dynamics remains unclear. Examining the microcircuits that participate in interictal spikes is thus an important first step towards addressing this issue, as the function of these transient synchronizations in either promoting or prohibiting seizures is currently under debate. To identify the microcircuits recruited in spontaneous interictal spikes in the absence of any proconvulsive drug or anaesthetic agent, we combine a chronic model of epilepsy with in vivo two-photon calcium imaging and multiunit extracellular recordings to map cellular recruitment within large populations of CA1 neurons in mice free to run on a self-paced treadmill. We show that GABAergic neurons, as opposed to their glutamatergic counterparts, are preferentially recruited during spontaneous interictal activity in the CA1 region of the epileptic mouse hippocampus. Although the specific cellular dynamics of interictal spikes are found to be highly variable, they are consistently associated with the activation of GABAergic neurons, resulting in a perisomatic inhibitory restraint that reduces neuronal spiking in the principal cell layer. Given the role of GABAergic neurons in shaping brain activity during normal cognitive function, their aberrant unbalanced recruitment during these transient events could have important downstream effects with clinical implications.

Normotopic cortex is the major contributor to epilepsy in experimental double cortex.

Subcortical band heterotopia (SBH) is a cortical malformation formed when neocortical neurons prematurely stop their migration in the white matter, forming a heterotopic band below the normotopic cortex, and is generally associated with intractable epilepsy. Although it is clear that the band heterotopia and the overlying cortex both contribute to creating an abnormal circuit prone to generate epileptic discharges, it is less understood which part of this circuitry is the most critical. Here, we sought to identify the origin of epileptiform activity in a targeted genetic model of SBH in rats.

Development of early-born γ-Aminobutyric acid hub neurons in mouse hippocampus from embryogenesis to adulthood

Early‐born γ‐aminobutyric acid (GABA) neurons (EBGNs) are major components of the hippocampal circuit because at early postnatal stages they form a subpopulation of “hub cells” transiently supporting CA3 network synchronization (Picardo et al. [2011] Neuron 71:695–709). It is therefore essential to determine when these cells acquire the remarkable morphofunctional attributes supporting their network function and whether they develop into a specific subtype of interneuron into adulthood. Inducible genetic fate mapping conveniently allows for the labeling of EBGNs throughout their life. EBGNs were first analyzed during the perinatal week. We observed that EBGNs acquired mature characteristics at the time when the first synapse‐driven synchronous activities appeared in the form of giant depolarizing potentials. The fate of EBGNs was next analyzed in the adult hippocampus by using anatomical characterization. Adult EBGNs included a significant proportion of cells projecting selectively to the septum; in turn, EBGNs were targeted by septal and entorhinal inputs. In addition, most EBGNs were strongly targeted by cholinergic and monoaminergic terminals, suggesting significant subcortical innervation. Finally, we found that some EBGNs located in the septum or the entorhinal cortex also displayed a long‐range projection that we traced to the hippocampus. Therefore, this study shows that the maturation of the morphophysiological properties of EBGNs mirrors the evolution of early network dynamics, suggesting that both phenomena may be causally linked. We propose that a subpopulation of EBGNs forms into adulthood a scaffold of GABAergic projection neurons linking the hippocampus to distant structures. J. Comp. Neurol. 524:2440–2461, 2016.

Image-based adaptive optics for in vivo imaging in the hippocampus

Adaptive optics is a promising technique for the improvement of microscopy in tissues. A large palette of indirect and direct wavefront sensing methods has been proposed for in vivo imaging in experimental animal models. Application of most of these methods to complex samples suffers from either intrinsic and/or practical difficulties. Here we show a theoretically optimized wavefront correction method for inhomogeneously labeled biological samples. We demonstrate its performance at a depth of 200 μm in brain tissue within a sparsely labeled region such as the pyramidal cell layer of the hippocampus, with cells expressing GCamP6. This method is designed to be sample-independent thanks to an automatic axial locking on objects of interest through the use of an image-based metric that we designed. Using this method, we show an increase of in vivo imaging quality in the hippocampus.

Modeling driver cells in developing neuronal networks

Spontaneous emergence of synchronized population activity is a characteristic feature of developing brain circuits. Recent experiments in the developing neo-cortex showed the existence of driver cells able to impact the synchronization dynamics when single-handedly stimulated. We have developed a spiking network model capable to reproduce the experimental results, thus identifying two classes of driver cells: functional hubs and low functionally connected (LC) neurons. The functional hubs arranged in a clique orchestrated the synchronization build-up, while the LC drivers were lately or not at all recruited in the synchronization process. Notwithstanding, they were able to alter the network state when stimulated by modifying the temporal activation of the functional clique or even its composition. LC drivers can lead either to higher population synchrony or even to the arrest of population dynamics, upon stimulation. Noticeably, some LC driver can display both effects depending on the received stimulus. We show that in the model the presence of inhibitory neurons together with the assumption that younger cells are more excitable and less connected is crucial for the emergence of LC drivers. These results provide a further understanding of the structural-functional mechanisms underlying synchronized firings in developing circuits possibly related to the coordinated activity of cell assemblies in the adult brain. Author Summary There is timely interest on the impact of peculiar neurons (driver cells) and of small neuronal sub-networks (cliques) on operational brain dynamics. We first provide experimental data concerning the effect of stimulated driver cells on the bursting activity observable in the developing entorhinal cortex. Secondly, we develop a network model able to fully reproduce the experimental observations. Analogously to the experiments two types of driver cells can be identified: functional hubs and low functionally connected (LC) drivers. We explain the role of hub neurons, arranged in a clique, for the orchestration of the bursting activity in control conditions. Furthermore, we report a new mechanism, which can explain why and how LC drivers emerge in the structural-functional organization of the enthorinal cortex.

Adaptive optics for in vivo two-photon calcium imaging of neuronal networks

The landscape of biomedical research in neuroscience has changed dramatically in recent years as a result of spectacular progress in dynamic microscopy. However, the optical accessibility of deep brain structures or deeper regions of the surgically exposed hippocampus (a few 100 microns typically) remains limited, due to volumic aberrations created by the sample inhomogeneities. Adaptive optics can correct for these aberrations. Our goal is to realize a novel adaptive optics module dedicated to in vivo two-photon calcium imaging of the hippocampus. The key issue in adaptive optics is the ability to perform an accurate and reliable wavefront sensing. In two- photon microscopy indirect methods are required. Two families of approaches have been proposed so far, the modal sensorless technique and a method based on pupil segmentation. We present here a formal comparison of these approaches, in particular as a function of the amount of aberrations.