18,890 research outputs found
Two-photon imaging and analysis of neural network dynamics
The glow of a starry night sky, the smell of a freshly brewed cup of coffee
or the sound of ocean waves breaking on the beach are representations of the
physical world that have been created by the dynamic interactions of thousands
of neurons in our brains. How the brain mediates perceptions, creates thoughts,
stores memories and initiates actions remains one of the most profound puzzles
in biology, if not all of science. A key to a mechanistic understanding of how
the nervous system works is the ability to analyze the dynamics of neuronal
networks in the living organism in the context of sensory stimulation and
behaviour. Dynamic brain properties have been fairly well characterized on the
microscopic level of individual neurons and on the macroscopic level of whole
brain areas largely with the help of various electrophysiological techniques.
However, our understanding of the mesoscopic level comprising local populations
of hundreds to thousands of neurons (so called 'microcircuits') remains
comparably poor. In large parts, this has been due to the technical
difficulties involved in recording from large networks of neurons with
single-cell spatial resolution and near- millisecond temporal resolution in the
brain of living animals. In recent years, two-photon microscopy has emerged as
a technique which meets many of these requirements and thus has become the
method of choice for the interrogation of local neural circuits. Here, we
review the state-of-research in the field of two-photon imaging of neuronal
populations, covering the topics of microscope technology, suitable fluorescent
indicator dyes, staining techniques, and in particular analysis techniques for
extracting relevant information from the fluorescence data. We expect that
functional analysis of neural networks using two-photon imaging will help to
decipher fundamental operational principles of neural microcircuits.Comment: 36 pages, 4 figures, accepted for publication in Reports on Progress
in Physic
A Bayesian approach for inferring neuronal connectivity from calcium fluorescent imaging data
Deducing the structure of neural circuits is one of the central problems of
modern neuroscience. Recently-introduced calcium fluorescent imaging methods
permit experimentalists to observe network activity in large populations of
neurons, but these techniques provide only indirect observations of neural
spike trains, with limited time resolution and signal quality. In this work we
present a Bayesian approach for inferring neural circuitry given this type of
imaging data. We model the network activity in terms of a collection of coupled
hidden Markov chains, with each chain corresponding to a single neuron in the
network and the coupling between the chains reflecting the network's
connectivity matrix. We derive a Monte Carlo Expectation--Maximization
algorithm for fitting the model parameters; to obtain the sufficient statistics
in a computationally-efficient manner, we introduce a specialized
blockwise-Gibbs algorithm for sampling from the joint activity of all observed
neurons given the observed fluorescence data. We perform large-scale
simulations of randomly connected neuronal networks with biophysically
realistic parameters and find that the proposed methods can accurately infer
the connectivity in these networks given reasonable experimental and
computational constraints. In addition, the estimation accuracy may be improved
significantly by incorporating prior knowledge about the sparseness of
connectivity in the network, via standard L penalization methods.Comment: Published in at http://dx.doi.org/10.1214/09-AOAS303 the Annals of
Applied Statistics (http://www.imstat.org/aoas/) by the Institute of
Mathematical Statistics (http://www.imstat.org
A roadmap to integrate astrocytes into Systems Neuroscience.
Systems neuroscience is still mainly a neuronal field, despite the plethora of evidence supporting the fact that astrocytes modulate local neural circuits, networks, and complex behaviors. In this article, we sought to identify which types of studies are necessary to establish whether astrocytes, beyond their well-documented homeostatic and metabolic functions, perform computations implementing mathematical algorithms that sub-serve coding and higher-brain functions. First, we reviewed Systems-like studies that include astrocytes in order to identify computational operations that these cells may perform, using Ca2+ transients as their encoding language. The analysis suggests that astrocytes may carry out canonical computations in a time scale of subseconds to seconds in sensory processing, neuromodulation, brain state, memory formation, fear, and complex homeostatic reflexes. Next, we propose a list of actions to gain insight into the outstanding question of which variables are encoded by such computations. The application of statistical analyses based on machine learning, such as dimensionality reduction and decoding in the context of complex behaviors, combined with connectomics of astrocyte-neuronal circuits, is, in our view, fundamental undertakings. We also discuss technical and analytical approaches to study neuronal and astrocytic populations simultaneously, and the inclusion of astrocytes in advanced modeling of neural circuits, as well as in theories currently under exploration such as predictive coding and energy-efficient coding. Clarifying the relationship between astrocytic Ca2+ and brain coding may represent a leap forward toward novel approaches in the study of astrocytes in health and disease
Real-time optical manipulation of cardiac conduction in intact hearts
Optogenetics has provided new insights in cardiovascular research, leading to new methods for cardiac pacing, resynchronization therapy and cardioversion. Although these interventions have clearly demonstrated the feasibility of cardiac manipulation, current optical stimulation strategies do not take into account cardiac wave dynamics in real time. Here, we developed an allâoptical platform complemented by integrated, newly developed software to monitor and control electrical activity in intact mouse hearts. The system combined a wideâfield mesoscope with a digital projector for optogenetic activation. Cardiac functionality could be manipulated either in freeârun mode with submillisecond temporal resolution or in a closedâloop fashion: a tailored hardware and software platform allowed realâtime intervention capable of reacting within 2 ms. The methodology was applied to restore normal electrical activity after atrioventricular block, by triggering the ventricle in response to optically mapped atrial activity with appropriate timing. Realâtime intraventricular manipulation of the propagating electrical wavefront was also demonstrated, opening the prospect for realâtime resynchronization therapy and cardiac defibrillation. Furthermore, the closedâloop approach was applied to simulate a reâentrant circuit across the ventricle demonstrating the capability of our system to manipulate heart conduction with high versatility even in arrhythmogenic conditions. The development of this innovative optical methodology provides the first proofâofâconcept that a realâtime optically based stimulation can control cardiac rhythm in normal and abnormal conditions, promising a new approach for the investigation of the (patho)physiology of the heart
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Network Properties Revealed during Multi-Scale Calcium Imaging of Seizure Activity in Zebrafish.
Seizures are characterized by hypersynchronization of neuronal networks. Understanding these networks could provide a critical window for therapeutic control of recurrent seizure activity, i.e., epilepsy. However, imaging seizure networks has largely been limited to microcircuits in vitro or small "windows" in vivo. Here, we combine fast confocal imaging of genetically encoded calcium indicator (GCaMP)-expressing larval zebrafish with local field potential (LFP) recordings to study epileptiform events at whole-brain and single-neuron levels in vivo. Using an acute seizure model (pentylenetetrazole, PTZ), we reliably observed recurrent electrographic ictal-like events associated with generalized activation of all major brain regions and uncovered a well-preserved anterior-to-posterior seizure propagation pattern. We also examined brain-wide network synchronization and spatiotemporal patterns of neuronal activity in the optic tectum microcircuit. Brain-wide and single-neuronal level analysis of PTZ-exposed and 4-aminopyridine (4-AP)-exposed zebrafish revealed distinct network dynamics associated with seizure and non-seizure hyperexcitable states, respectively. Neuronal ensembles, comprised of coactive neurons, were also uncovered during interictal-like periods. Taken together, these results demonstrate that macro- and micro-network calcium motifs in zebrafish may provide a greater understanding of epilepsy
Optical excitation and detection of neuronal activity
Optogenetics has emerged as an exciting tool for manipulating neural
activity, which in turn, can modulate behavior in live organisms. However,
detecting the response to the optical stimulation requires electrophysiology
with physical contact or fluorescent imaging at target locations, which is
often limited by photobleaching and phototoxicity. In this paper, we show that
phase imaging can report the intracellular transport induced by optogenetic
stimulation. We developed a multimodal instrument that can both stimulate cells
with high spatial resolution and detect optical pathlength changes with
nanometer scale sensitivity. We found that optical pathlength fluctuations
following stimulation are consistent with active organelle transport.
Furthermore, the results indicate a broadening in the transport velocity
distribution, which is significantly higher in stimulated cells compared to
optogenetically inactive cells. It is likely that this label-free, contactless
measurement of optogenetic response will provide an enabling approach to
neuroscience.Comment: 20 pages, 5 figure
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Simultaneous mesoscopic and two-photon imaging of neuronal activity in cortical circuits.
Spontaneous and sensory-evoked activity propagates across varying spatial scales in the mammalian cortex, but technical challenges have limited conceptual links between the function of local neuronal circuits and brain-wide network dynamics. We present a method for simultaneous cellular-resolution two-photon calcium imaging of a local microcircuit and mesoscopic widefield calcium imaging of the entire cortical mantle in awake mice. Our multi-scale approach involves a microscope with an orthogonal axis design where the mesoscopic objective is oriented above the brain and the two-photon objective is oriented horizontally, with imaging performed through a microprism. We also introduce a viral transduction method for robust and widespread gene delivery in the mouse brain. These approaches allow us to identify the behavioral state-dependent functional connectivity of pyramidal neurons and vasoactive intestinal peptide-expressing interneurons with long-range cortical networks. Our imaging system provides a powerful strategy for investigating cortical architecture across a wide range of spatial scales
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