Controlling Bursting in Cortical Cultures with Closed-Loop Multi-Electrode Stimulation

Wagenaar, Daniel A.; Madhavan, Radhika; Pine, J.; Potter, Steve M.

doi:10.1523/jneurosci.4209-04.2005

Cited by 395 publications

(391 citation statements)

References 81 publications

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“…The observation that spike activity increased indicates that the added astroglia did not significantly occlude electrodes from their contact with neurons. Another issue of cell density of 500 cells mm −2 might be considered high for many studies using intracellular recording, but is much lower than in the brain and lower than the 1000-2500 cells mm −2 used by Potter's group (Wagenaar et al, 2005;Wagenaar et al, 2006).…”

Section: Discussionmentioning

confidence: 91%

Added astroglia promote greater synapse density and higher activity in neuronal networks

2007

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Astroglia are known to potentiate individual synapses, but their contribution to networks is unclear.Here we examined the effect of adding either astroglia or media conditioned by astroglia on entire networks of rat hippocampal neurons cultured on microelectrode arrays. Added astroglia increased spontaneous spike rates nearly two-fold and glutamate-stimulated spiking by six-fold, with desensitization eliminated for bath addition of 25 μM glutamate. Astrocyte-conditioned medium partly mimicked the effects of added astroglia. Bursting behavior was largely unaffected by added astroglia except with added glutamate. Addition of the GABA A receptor antagonist bicuculline also increased spike rates but with more subtle differences between networks without or with added astroglia. This indicates that networks without added astroglia were inhibited greatly. In all conditions, the log-log distribution of spike rates fit well to linear distributions over three orders of magnitude. Networks with added astroglia shifted consistently toward higher spike rates. Immunostaining for GFAP revealed a linear increase with added astroglia, which also increased neuronal survival. The increased spike rates with added astroglia correlated with a 1.7-fold increase in immunoreactive synaptophysin puncta, and increases of six-fold for GABA Aβ , two-fold for NMDA-R1 and two-fold for Glu-R1 puncta, with receptor clustering that indicated synaptic scaling. Together, these results indicate that added astroglia increase the density of synapses and receptors, and facilitate higher spike rates for many elements in the network. These effects are reproduced by glia-conditioned media, with the exception of glutamate-mediated transmission.

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Section: Discussionmentioning

confidence: 91%

Added astroglia promote greater synapse density and higher activity in neuronal networks

2007

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“…Of relevance to this work, simulations of central nervous system neurons in the context of the cortex as well as the spinal cord have also been performed Grill 1999, McIntyre andGrill 2002), and indicate along with experimental studies that the primary location of action potential (AP) generation induced by applied electric fields is at the axon initial segment Bullier 1998a, Nowak andBullier 1998b). Of interest, recent reduced experimental models examining planar patterns of cultured neurons grown over a microelectrode array also exhibit reductions and cessations in bursting behaviour for a variety of imposed stimulation fields (Wagenaar et al 2005). …”

Section: Introductionmentioning

confidence: 99%

Studies of stimulus parameters for seizure disruption using neural network simulations

et al. 2007

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A large scale neural network simulation with realistic cortical architecture has been undertaken to investigate the effects of external electrical stimulation on the propagation and evolution of ongoing seizure activity. This is an effort to explore the parameter space of stimulation variables to uncover promising avenues of research for this therapeutic modality. The model consists of an approximately 800 μm × 800 μm region of simulated cortex, and includes seven neuron classes organized by cortical layer, inhibitory or excitatory properties, and electrophysiological characteristics. The cell dynamics are governed by a modified version of the Hodgkin-Huxley equations in single compartment format. Axonal connections are patterned after histological data and published models of local cortical wiring. Stimulation induced action potentials take place at the axon initial segments, according to threshold requirements on the applied electric field distribution. Stimulation induced action potentials in horizontal axonal branches are also separately simulated. The calculations are performed on a 16 node distributed 32-bit processor system. Clear differences in seizure evolution are presented for stimulated versus the undisturbed rhythmic activity. Data is provided for frequency dependent stimulation effects demonstrating a plateau effect of stimulation efficacy as the applied frequency is increased from 60 Hz to 200 Hz. Timing of the stimulation with respect to the underlying rhythmic activity demonstrates a phase dependent sensitivity. Electrode height and position effects are also presented. Using a dipole stimulation electrode arrangement, clear orientation effects of the dipole with respect to the model connectivity is also demonstrated. A sensitivity analysis of these results as a function of the stimulation threshold is also provided.

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“…Multi-electrode arrays (MEAs) have also been extensively used to probe cultured neural networks (Corner et al, 2002;Pasquale et al, 2008;Wagenaar et al, 2005), namely to characterize the bursting activity in developing cultured neural networks and control activity in these networks by stimulating at different electrode sites (Madhavan et al, 2006;Massobrio et al, 2007;Wagenaar et al, 2005). However, the extracellular stimulation from the array suffers from poor spatial localization (Heuschkel et al, 2002) and stimulation artifacts (Wagenaar and Potter, 2004).…”

Section: Discussionmentioning

confidence: 99%

Laser-evoked synaptic transmission in cultured hippocampal neurons expressing channelrhodopsin-2 delivered by adeno-associated virus

Wang¹,

Hasan²,

Seung³

2009

Journal of Neuroscience Methods

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We present a method for studying synaptic transmission in mass cultures of dissociated hippocampal neurons based on patch clamp recording combined with laser stimulation of neurons expressing Channelrhodopsin-2 (ChR2). Our goal was to use the high spatial resolution of laser illumination to come as close as possible to the ideal of identifying monosynaptically coupled pairs of neurons, which is conventionally done using microisland rather than mass cultures. Using recombinant adenoassociated virus (rAAV) to deliver the ChR2 gene, we focused on the time period between 14 and 20 days in vitro, during which expression levels are high, and spontaneous bursting activity has not yet started. Stimulation by wide-field illumination is sufficient to make the majority of ChR2-expressing neurons spike. Stimulation with a laser spot at least 10 μm in diameter also produces action potentials, but in a reduced fraction of neurons. We studied synaptic transmission by voltageclamping a neuron with low expression of ChR2 and scanning a 40 μm laser spot at surrounding locations. Responses were observed to stimulation at a subset of locations in the culture, indicating spatial localization of stimulation. Pharmacological means were used to identify responses that were synaptic. Many responses were of smaller amplitude than those typically found in microisland cultures. We were unable to find an entirely reliable criterion for distinguishing between monosynaptic and polysynaptic responses. However, we propose that postsynaptic currents with Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. HHMI Author ManuscriptHHMI Author Manuscript HHMI Author Manuscript small amplitudes, simple shapes, and latencies not much greater than 8 msec are reasonable candidates for monosynaptic interactions.

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Controlling Bursting in Cortical Cultures with Closed-Loop Multi-Electrode Stimulation

Cited by 395 publications

References 81 publications

Added astroglia promote greater synapse density and higher activity in neuronal networks

Added astroglia promote greater synapse density and higher activity in neuronal networks

Studies of stimulus parameters for seizure disruption using neural network simulations

Laser-evoked synaptic transmission in cultured hippocampal neurons expressing channelrhodopsin-2 delivered by adeno-associated virus

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