A Micro-Electrode Array device coupled to a laser-based system for the local stimulation of neurons by optical release of glutamate

Ghezzi, Diego; Menegon, Andrea; Pedrocchi, Alessandra; Valtorta, Flavia; Ferrigno, Giancarlo

doi:10.1016/j.jneumeth.2008.08.003

Cited by 28 publications

(25 citation statements)

References 24 publications

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“…This is particularly problematic when functionally different cells lie in close spatial proximity. For these reasons we seek reliable alternatives to electrode stimulation for artifact-free, controllable, and specific cell stimulation [27,28]. Promising results were achieved using 20 light triggerable ion channels that depolarize a cell and subsequently cause action potentials [29,30,31].…”

Section: Introductionmentioning

confidence: 99%

Light induced stimulation and delay of cardiac activity

et al. 2010

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Section: Introductionmentioning

confidence: 99%

Light induced stimulation and delay of cardiac activity

et al. 2010

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“…However, the extracellular stimulation from the array suffers from poor spatial localization (Heuschkel et al, 2002) and stimulation artifacts (Wagenaar and Potter, 2004). These stimulation issues have been circumvented by using optical stimulation of caged glutamate to stimulate neurons, while recording exracellular signals via the MEA (Ghezzi et al, 2008). Laser stimulation of neurons expressing ChR2 could potentially be used in a similar fashion, with the advantage that there would be no accumulation of glutamate in the bath.…”

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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“…Moreover, also external devices such as optical fibres (Bernardinelli et al, 2005) or semiconductor UV light-emitting diodes (Venkataramani et al, 2007) have been used. The idea of coupling MEAs and optical uncaging has been explored (Ghezzi et al, 2008) using a micro-actuated optical fibre that is able to activate a single site of a cultured neuronal network. In that work, the evaluation of the compound diffusion and of the uncaging efficiency confirmed the applicability of this approach to the local excitation of a selected region of a neuronal network cultured on a MEA device.…”

Section: Scientific Backgroundmentioning

confidence: 99%

“…In comparison to electrical stimulation, where the electrical stimulus spreads over the whole biological preparation with an amplitude decreasing with the square of the distance from the stimulation site (Heuschkel et al, 2002), the optical stimulation scheme of caged compounds is limited to areas that are exposed to light pulses with sufficient energy to uncage the compounds and diffusion of the compounds in the medium, i.e. uncaging takes place only in a well defined volume (Ghezzi et al, 2008). In order to allow local chemical stimulation, the spatial control of light, i.e.…”

Section: Evaluation Of Photostimulation With Photomeamentioning

confidence: 99%