Fast Voltage-sensitive Dye Recording of Membrane Potential Changes at Multiple Sites on an Individual Nerve Cell in the Rat Cortical Slice

Antic, Srdjan D.; Major, Guy; Chen, W. R.; Wuskel, J.; Loew, Leslie M.; Zečević, Dejan

doi:10.1086/bblv193n2p261

Cited by 11 publications

(8 citation statements)

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“…Actually, bath application of JPW3027 (originally engineered for intracellular application) produced less background than RH795, a VSD engineered for extracellular application. VSDs bind indiscriminately to any lipid membrane [25] and we believe that the higher hydrophobicity of JPW3027 compared to RH795 causes a lesser affinity to lipids, producing a ‘cleaner’ labeling. While few studies so far have used VSDs to look at dissociated cells they commonly have been used in monitoring neuronal activity in slice preparations and in vivo [26].…”

Section: Discussionmentioning

confidence: 94%

“…Moreover, changes in [Ca 2+ ] are not linearly related to membrane potential variation and diverse Ca 2+ buffering properties of different sub-cellular compartments further complicate the relationship between [Ca 2+ ] and membrane potential. On the other hand, several VSDs are capable to resolve changes in membrane potential with a linear variation in fluorescence [25] and in the sub-millisecond range [6], [27].…”

Section: Discussionmentioning

confidence: 99%

“…Also, VSD changes in Δ F / F 0 in response to an AP are at least an order of magnitude smaller than changes reported by Ca 2+ indicators [20], [25]. In order to improve the signal-to-noise ratio, other groups have averaged several image acquisition sweeps [27].…”

Section: Discussionmentioning

confidence: 99%

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A Voltage-Sensitive Dye-Based Assay for the Identification of Differentiated Neurons Derived from Embryonic Neural Stem Cell Cultures

et al. 2010

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BackgroundPluripotent and multipotent stem cells hold great therapeutical promise for the replacement of degenerated tissue in neurological diseases. To fulfill that promise we have to understand the mechanisms underlying the differentiation of multipotent cells into specific types of neurons. Embryonic stem cell (ESC) and embryonic neural stem cell (NSC) cultures provide a valuable tool to study the processes of neural differentiation, which can be assessed using immunohistochemistry, gene expression, Ca2+-imaging or electrophysiology. However, indirect methods such as protein and gene analysis cannot provide direct evidence of neuronal functionality. In contrast, direct methods such as electrophysiological techniques are well suited to produce direct evidence of neural functionality but are limited to the study of a few cells on a culture plate.Methodology/Principal FindingsIn this study we describe a novel method for the detection of action potential-capable neurons differentiated from embryonic NSC cultures using fast voltage-sensitive dyes (VSD). We found that the use of extracellularly applied VSD resulted in a more detailed labeling of cellular processes compared to calcium indicators. In addition, VSD changes in fluorescence translated precisely to action potential kinetics as assessed by the injection of simulated slow and fast sodium currents using the dynamic clamp technique. We further demonstrate the use of a finite element model of the NSC culture cover slip for optimizing electrical stimulation parameters.Conclusions/SignificanceOur method allows for a repeatable fast and accurate stimulation of neurons derived from stem cell cultures to assess their differentiation state, which is capable of monitoring large amounts of cells without harming the overall culture.

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Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

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A Voltage-Sensitive Dye-Based Assay for the Identification of Differentiated Neurons Derived from Embryonic Neural Stem Cell Cultures

et al. 2010

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“…Initial experiments used pressure ejection from sharp microelectrodes, again in invertebrate neurons [4,55]. Subsequent experiments applied voltage-sensitive dyes internally via diffusion from the recording pipette during patch-clamp recordings from neurons in brain slices [1,2]. The use of the patchclamp technique increased the ability to passively fill neurons with voltage-sensitive dyes due to the larger tip diameter of patch pipettes compared to sharp microelectrodes.…”

Section: Voltage Imaging Using Internally Applied Dyesmentioning

confidence: 99%

Imaging membrane potential in dendrites and axons of single neurons

Stuart

Palmer

2006

Pflugers Arch - Eur J Physiol

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This review focuses on the use of imaging techniques to record electrical signaling in the fine processes of neurons such as dendrites and axons. Voltage imaging began with the use and development of externally applied voltage-sensitive dyes. With the introduction of internally applied dyes and advances in detection technology, it is now possible to record supra-threshold action potential responses, as well as sub-threshold synaptic potentials, in fine neuronal processes including dendritic spines. The development of genetically coded sensors, as well as variants of laser scanning microscopy such as second harmonic generation, offers promise for further advances in this field. Through the use and further development of these methods, optical imaging of membrane potential will continue to be a valuable tool for investigators wishing to explore the electrical events underlying single neuronal computation.

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“…Acute brain preparations and slices are commonly used for in vitro study of mammalian brain functions (Ballanyi, 1999) and optical imaging (Cohen, 1988;Canepari and Zecevic, 2010). Water-soluble fluorescent ANEP-based VSDs, like di-1-ANEPPS (JPW3028), di-2-ANEPEQ (JPW1114) or di-4-ANEPPS, are often chosen for intracellular loading and bath-application in cortical slices (Grinvald et al, 1987;Antic et al, 1997;Foust et al, 2011;Palmer and Stuart, 2006;Sinha and Saggau, 1999;Tominaga et al, 2000). However, a series of lipophilic VSDs with significantly higher sensitivity to membrane potential changes has been synthesized .…”

mentioning

confidence: 99%

Biolistic delivery of voltage-sensitive dyes for fast recording of membrane potential changes in individual neurons in rat brain slices

Aseyev

Roshchin

Ierusalimsky

et al. 2013

Journal of Neuroscience Methods

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Fast Voltage-sensitive Dye Recording of Membrane Potential Changes at Multiple Sites on an Individual Nerve Cell in the Rat Cortical Slice

Cited by 11 publications

References 0 publications

A Voltage-Sensitive Dye-Based Assay for the Identification of Differentiated Neurons Derived from Embryonic Neural Stem Cell Cultures

A Voltage-Sensitive Dye-Based Assay for the Identification of Differentiated Neurons Derived from Embryonic Neural Stem Cell Cultures

Imaging membrane potential in dendrites and axons of single neurons

Biolistic delivery of voltage-sensitive dyes for fast recording of membrane potential changes in individual neurons in rat brain slices

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