Functional multineuron calcium imaging for systems pharmacology

Takahara, Yukio; Ishikawa, Daisuke; Matsuki, Norio; Ikegaya, Yuji

doi:10.1007/s00216-010-3740-6

Cited by 11 publications

(9 citation statements)

References 58 publications

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“…Our experiments fall within the context of functional multineuron calcium imaging (fMCI), a technique based in the ability to examine network activity in large neuronal populations and with single-cell resolution (Stosiek et al, 2003; Ohki et al, 2005; Bonifazi et al, 2009; Takahashi et al, 2010a,b). fMCI has received substantial attention in the last years driven by the spectacular development of optogenetic tools and genetically encoded calcium indicators, which allow to monitor and probe neuronal circuits in vivo without the need of electrodes or other invasive measuring techniques (Yizhar et al, 2011a).…”

Section: Discussionmentioning

confidence: 99%

Identification of neuronal network properties from the spectral analysis of calcium imaging signals in neuronal cultures

Tibau

Valencia

Soriano

2013

Front. Neural Circuits

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Neuronal networks in vitro are prominent systems to study the development of connections in living neuronal networks and the interplay between connectivity, activity and function. These cultured networks show a rich spontaneous activity that evolves concurrently with the connectivity of the underlying network. In this work we monitor the development of neuronal cultures, and record their activity using calcium fluorescence imaging. We use spectral analysis to characterize global dynamical and structural traits of the neuronal cultures. We first observe that the power spectrum can be used as a signature of the state of the network, for instance when inhibition is active or silent, as well as a measure of the network's connectivity strength. Second, the power spectrum identifies prominent developmental changes in the network such as GABAA switch. And third, the analysis of the spatial distribution of the spectral density, in experiments with a controlled disintegration of the network through CNQX, an AMPA-glutamate receptor antagonist in excitatory neurons, reveals the existence of communities of strongly connected, highly active neurons that display synchronous oscillations. Our work illustrates the interest of spectral analysis for the study of in vitro networks, and its potential use as a network-state indicator, for instance to compare healthy and diseased neuronal networks.

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Section: Discussionmentioning

confidence: 99%

Identification of neuronal network properties from the spectral analysis of calcium imaging signals in neuronal cultures

Tibau

Valencia

Soriano

2013

Front. Neural Circuits

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“…Calcium imaging is an established technique for monitoring neuronal activity 6,18,21 and can be effectively used to increase the throughput of simultaneously acquired signals. 22,23 Some studies have even shown that it is possible to detect single action potentials from captured Ca 2+ signals. 23 Similar calcium imaging techniques have previously been used within microfluidic devices to examine how a cell responds to locally applied chemical insults, 14 as well as in cortico-striatal co-cultures to construct orientated synaptic networks.…”

Section: Calcium Imaging and Synaptic Communicationmentioning

confidence: 99%

“…22,23 Some studies have even shown that it is possible to detect single action potentials from captured Ca 2+ signals. 23 Similar calcium imaging techniques have previously been used within microfluidic devices to examine how a cell responds to locally applied chemical insults, 14 as well as in cortico-striatal co-cultures to construct orientated synaptic networks. 21 However, while these studies demonstrate the use of calcium imaging as a method for monitoring cell activity within microfluidic devices, the technique was not used in a dynamic assay to show how one stimulated culture can synaptically influence the activity of the connected culture.…”

Section: Calcium Imaging and Synaptic Communicationmentioning

confidence: 99%

Chemically induced synaptic activity between mixed primary hippocampal co-cultures in a microfluidic system

Robertson

Bushell

Zagnoni

2014

Integrative Biology

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Primary neuronal cultures are an invaluable in vitro tool for examining the fundamental physiological changes that occur in diseases of the central nervous system. In this work, we have used a microfluidic device to grow twin cultures of primary hippocampal neuronal/glia cells which are synaptically connected but environmentally isolated. Immunocytochemical staining, for β-III-Tubulin and synaptophysin, indicated that the two neuronal populations were physically connected and that synapses were present. By dispensing predefined volumes of fluids into the device inlets, one culture was chemically stimulated and the consequent increase in neuronal activity in the opposing culture was monitored using calcium imaging. To optimise the experimental procedures, we validated a numerical model that estimates the concentration distribution of substances under dynamic fluidic conditions, proposing that no cross contamination of chemical stimuli occurred during the experiments. Calcium imaging and local chemical stimulation were used to confirm synaptic connectivity between the cultures. Chemical stimulation of one population, using KCl or glutamate, resulted in a significant increase of calcium events in both neurons and astrocytes of the connected population. The integration of the system and techniques described here presents a novel methodology for probing the functional synaptic connectivity between mixed primary hippocampal co-cultures, creating an in vitro testing platform for the high-throughput investigation of synaptic activity modulation either by novel compounds or in in vitro disease models.

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“…To address this issue, functional multineuron Ca 2+ imaging (fMCI) has been developed as a technique for neural network imaging at single‐cell resolution in recent decades (Smetters et al ., ; Cossart et al ., ; Takahashi et al ., , ). Many studies have made use of fMCI to probe the dynamics of neuronal circuits (Cossart et al ., ; Ikegaya et al ., ; Sasaki et al ., ; Bonifazi et al ., ).…”

Section: Introductionmentioning

confidence: 99%

Heterogeneous effects of antiepileptic drugs in an in vitro epilepsy model – a functional multineuron calcium imaging study

Hongo

Takasu

Ikegaya

et al. 2015

Eur J of Neuroscience

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Epilepsy is a chronic brain disease characterised by recurrent seizures. Many studies of this disease have focused on local neuronal activity, such as local field potentials in the brain. In addition, several recent studies have elucidated the collective behavior of individual neurons in a neuronal network that emits epileptic activity. However, little is known about the effects of antiepileptic drugs on neuronal networks during seizure-like events (SLEs) at single-cell resolution. Using functional multineuron Ca(2+) imaging (fMCI), we monitored the activities of multiple neurons in the rat hippocampal CA1 region on treatment with the proconvulsant bicuculline under Mg(2+) -free conditions. Bicuculline induced recurrent synchronous Ca(2+) influx, and the events were correlated with SLEs. Other proconvulsants, such as 4-aminopyridine, pentetrazol, and pilocarpine, also induced synchronous Ca(2+) influx. We found that the antiepileptic drugs phenytoin, flupirtine, and ethosuximide, which have different mechanisms of action, exerted heterogeneous effects on bicuculline-induced synchronous Ca(2+) influx. Phenytoin and flupirtine significantly decreased the peak, the amount of Ca(2+) influx and the duration of synchronous events in parallel with the duration of SLEs, whereas they did not abolish the synchronous events themselves. Ethosuximide increased the duration of synchronous Ca(2+) influx and SLEs. Furthermore, the magnitude of the inhibitory effect of phenytoin on the peak synchronous Ca(2+) influx level differed according to the peak amplitude of the synchronous event in each individual cell. Evaluation of the collective behavior of individual neurons by fMCI seems to be a powerful tool for elucidating the profiles of antiepileptic drugs.

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Functional multineuron calcium imaging for systems pharmacology

Cited by 11 publications

References 58 publications

Identification of neuronal network properties from the spectral analysis of calcium imaging signals in neuronal cultures

Identification of neuronal network properties from the spectral analysis of calcium imaging signals in neuronal cultures

Chemically induced synaptic activity between mixed primary hippocampal co-cultures in a microfluidic system

Heterogeneous effects of antiepileptic drugs in an in vitro epilepsy model – a functional multineuron calcium imaging study

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