Optical Recording of Membrane Potential Changes From Single Neurons Using Intracellular Voltage-Sensitive Dyes

1991 ◽  
Vol 181 (2) ◽  
pp. 332-333
Author(s):  
D. Zecevic
Keyword(s):  
Membrane Potential ◽  
Optical Recording ◽  
Single Neurons ◽  
Voltage Sensitive Dyes

Optical recording of membrane potential responses from early embryonic chick ganglia using voltage-sensitive dyes

1985 ◽  
Vol 17 (1-2) ◽  
pp. 39-51 ◽  
Author(s):  
Tetsuro Sakai ◽  
Akihiko Hirota ◽  
Hitoshi Komuro ◽  
Shiroh Fujii ◽  
Kohtaro Kamino
Keyword(s):  
Membrane Potential ◽  
Optical Recording ◽  
Embryonic Chick ◽  
Voltage Sensitive Dyes

scholarly journals Fast Optical Recordings of Membrane Potential Changes From Dendrites of Pyramidal Neurons

1999 ◽  
Vol 82 (3) ◽  
pp. 1615-1621 ◽  
Author(s):  
Srdjan Antic ◽  
Guy Major ◽  
Dejan Zecevic
Keyword(s):  
Membrane Potential ◽  
Pyramidal Neurons ◽  
Spatial Dynamics ◽  
Brain Slices ◽  
Optical Recording ◽  
Neuronal Processes ◽  
Voltage Sensitive Dyes ◽  
Apical Dendrites ◽  
The Brain ◽  
Extracellular Environment

Understanding the biophysical properties of single neurons and how they process information is fundamental to understanding how the brain works. A technique that would allow recording of temporal and spatial dynamics of electrical activity in neuronal processes with adequate resolution would facilitate further research. Here, we report on the application of optical recording of membrane potential transients at many sites on neuronal processes of vertebrate neurons in brain slices using intracellular voltage-sensitive dyes. We obtained evidence that 1) loading the neurons with voltage-sensitive dye using patch electrodes is possible without contamination of the extracellular environment; 2) brain slices do not show any autofluorescence at the excitation/emission wavelengths used; 3) pharmacological effects of the dye were completely reversible; 4) the level of photodynamic damage already allows meaningful measurements and could be reduced further; 5) the sensitivity of the dye was comparable to that reported for invertebrate neurons; 6) the dye spread ∼500 μm into distal processes within 2 h incubation period. This distance should increase with longer incubation; 7) the optically recorded action potential signals from basolateral dendrites (that are difficult or impossible to approach by patch electrodes) and apical dendrites show that both direct soma stimulation and synaptic stimulation triggered action potentials that originated near the soma. The spikes backpropagated into both basolateral dendrites and apical processes; the propagation was somewhat faster in the apical dendrites.

Fluorescent potentiometric dyes for membrane-potential imaging

1994 ◽  
Vol 52 ◽  
pp. 166-167
Author(s):  
Leslie M. Loew
Keyword(s):  
Membrane Potential ◽  
Single Cells ◽  
Quantitative Imaging ◽  
Optical Recording ◽  
Biological Processes ◽  
Major Focus ◽  
The Past ◽  
Excitable Systems ◽  
Major Application ◽  
The Impact

A major application of potentiometric dyes has been the multisite optical recording of electrical activity in excitable systems. After being championed by L.B. Cohen and his colleagues for the past 20 years, the impact of this technology is rapidly being felt and is spreading to an increasing number of neuroscience laboratories. A second class of experiments involves using dyes to image membrane potential distributions in single cells by digital imaging microscopy - a major focus of this lab. These studies usually do not require the temporal resolution of multisite optical recording, being primarily focussed on slow cell biological processes, and therefore can achieve much higher spatial resolution. We have developed 2 methods for quantitative imaging of membrane potential. One method uses dual wavelength imaging of membrane-staining dyes and the other uses quantitative 3D imaging of a fluorescent lipophilic cation; the dyes used in each case were synthesized for this purpose in this laboratory.

Optical approaches to ontogeny of electrical activity and related functional organization during early heart development

1991 ◽  
Vol 71 (1) ◽  
pp. 53-91 ◽  
Author(s):  
K. Kamino
Keyword(s):  
Electrical Activity ◽  
Heart Development ◽  
Functional Organization ◽  
Embryonic Heart ◽  
Optical Recording ◽  
Additional Advantage ◽  
Physiological Functions ◽  
Spontaneous Electrical Activity ◽  
Somite Stage ◽  
Voltage Sensitive Dyes

Direct intracellular measurement of electrical events in the early embryonic heart is impossible because the cells are too small and frail to be impaled with microelectrodes; it is also not possible to apply conventional electrophysiological techniques to the early embryonic heart. For these reasons, complete understanding of the ontogeny of electrical activity and related physiological functions of the heart during early development has been hampered. Optical signals from voltage-sensitive dyes have provided a new powerful tool for monitoring changes in transmembrane voltage in a wide variety of living preparations. With this technique it is possible to make optical recordings from the cells that are inaccessible to microelectrodes. An additional advantage of the optical method for recording membrane potential activity is that electrical activity can be monitored simultaneously from many sites in a preparation. Thus, applying a multiple-site optical recording method with a 100- or 144-element photodiode array and voltage-sensitive dyes, we have been able to monitor, for the first time, spontaneous electrical activity in prefused cardiac primordia in the early chick embryos at the six- and the early seven-somite stages of development. We were able to determine that the time of initiation of the contraction is the middle period of the nine-somite stage. In the rat embryonic heart, the onset of spontaneous electrical activity and contraction occurs at the three-somite stage. In this review, a new view of the ontogenetic sequence of spontaneous electrical activity and related physiological functions such as ionic properties, pacemaker function, conduction, and characteristics of excitation-contraction coupling in the early embryonic heart are discussed.

Presynaptic Modulation of Synaptic Transmission by Pregnenolone Sulfate as Studied by Optical Recordings

2005 ◽  
Vol 94 (6) ◽  
pp. 4131-4144 ◽  
Author(s):  
Ling Chen ◽  
Masahiro Sokabe
Keyword(s):  
Synaptic Transmission ◽  
Glutamate Release ◽  
Hippocampal Slices ◽  
Receptor Activation ◽  
Optical Recording ◽  
Perforant Path ◽  
Dependent Manner ◽  
Pregnenolone Sulfate ◽  
Voltage Sensitive Dyes ◽  
Dose Dependent

The effects of pregnenolone sulfate (PREGS), a putative neurosteroid, on the transmission of perforant path–granule cell synapses were investigated with an optical recording technique in rat hippocampal slices stained with voltage-sensitive dyes. Application of PREGS to the bath solution resulted in an acute augmentation of EPSP in a dose-dependent manner. The PREGS effect was dependent on the extracellular Ca2+ concentration ([Ca2+]o), but independent of NMDA receptor activation. PREGS caused a decrease in paired-pulse facilitation, which implies that PREGS positively modulates presynaptic neurotransmitter releases. Firmer support for this mechanism was that PREGS augmented the synaptically induced glial depolarization (SIGD) that reflects the activity of electrogenic glutamate transporters in glial cells during the uptake of released glutamate. The selective α7nAChR antagonist α-BGT or MLA prevented the SIGD increase by PREGS. Furthermore DMXB, a selective α7nAChR agonist, mimicked the PREGS effect on SIGD and antagonized the effect of PREGS. The presynaptic effect of PREGS was partially attenuated by the L-type Ca2+ channel (VGCC) blocker nifedipine. Based on these findings, we proposed a novel mechanism underlying the facilitated synaptic transmission by PREGS: this neurosteroid sensitizes presynaptic α7nAChR that is followed by an activation of L-type VGCC to increase the presynaptic glutamate release.

Sub-Millisecond Time Resolved Imaging of Voltage Changes in Excitable Cells and Tissues Using Multiple Site Optical Recording of Transmembrane Voltage (Msortv) and Molecular Probes

1997 ◽  
Vol 3 (S2) ◽  
pp. 803-804
Author(s):  
B.M. Salzberg ◽  
A.L. Obaid
Keyword(s):  
Membrane Potential ◽  
Conformational Changes ◽  
Molecular Probes ◽  
Optical Recording ◽  
Membrane Voltage ◽  
Excitable Cells ◽  
Time Resolved ◽  
Molecular Weights ◽  
Transmembrane Voltage ◽  
Time Resolved Imaging

Molecular indicators of membrane potential may be used to obtain sub-millisecond time resolved images of transient changes in membrane voltage in a variety of biological systems. These probes are small amphipathic molecules having molecular weights of 400-500, and dimensions on the order of 10 Angstroms, which bind to, but do not cross cell membranes, and change either their absorbance or fluorescence in response to membrane voltage. These extrinsic optical signals depend linearly upon membrane potential, and the best of the dyes respond to a step change in voltage in less than 1.5 μsec at room temperature. The salient properties of fast potentiometric probes will be discussed, and the fidelity of optical recordings to transmembrane voltage changes will be considered.Since voltage changes in excitable cells take place on a time scale that is determined by the kinetics of conformational changes in membrane proteins, and by membrane electrical time constants, these changes tend to be very rapid, and resolving them requires imaging systems that are frequently orders of magnitude faster than usual video rates.

scholarly journals Mapping Membrane Potential Transients in Crayfish (Procambarus clarkii) Optic Lobe Neuropils With Voltage-Sensitive Dyes

1999 ◽  
Vol 81 (1) ◽  
pp. 334-344 ◽  
Author(s):  
Sergey Yagodin ◽  
Carlos Collin ◽  
Daniel L. Alkon ◽  
Norman F. Sheppard ◽  
David B. Sattelle
Keyword(s):  
Membrane Potential ◽  
Visual Information ◽  
Action Potentials ◽  
Optic Lobe ◽  
Procambarus Clarkii ◽  
Projection Neurons ◽  
Medulla Terminalis ◽  
Voltage Sensitive Dyes ◽  
Hemiellipsoid Body ◽  
Synaptic Responses

Yagodin, Sergey, Carlos Collin, Daniel L. Alkon, Norman F. Sheppard, Jr., and David B. Sattelle. Mapping membrane potential transients in crayfish ( Procambarus clarkii) optic lobe neuropils with voltage-sensitive dyes. J. Neurophysiol. 81: 334–344, 1999. Voltage-sensitive dyes NK 2761 and RH 155 were employed (in conjunction with a 12 × 12 photodiode array) to study membrane potential transients in optic lobe neuropils in the eye stalk of the crayfish Procambarus clarkii. By this means we investigated a pathway linking deutocerebral projection neurons, via hemiellipsoid body local interneurons, to an unidentified target (most likely neurons processing visual information) in the medulla terminalis. Rapid (10- to 20-ms duration), transient changes in absorption with the characteristics of action potentials were recorded from the optic nerve and the region occupied by deutocerebral projection neurons after stimulation of the olfactory globular tract in the optic nerve and were blocked by 1 μM tetrodotoxin. Action potentials appeared to propagate to the glomerular layer of the hemiellipsoid body where synaptic responses were recorded from a restricted region of the hemiellipsoid body occupied by dendrites of hemiellipsoid body neurons. Action potentials were also recorded from processes of hemiellipsoid body neurons located in the medulla terminalis. Synaptic responses in the hemiellipsoid body and medulla terminalis were eliminated by addition to the saline of 500 μM Cd2+ or 20 mM Co2+, whereas the action potential attributed to branches of deutocerebral projection neurons in the hemiellipsoid body remained unaffected. Action potentials of hemiellipsoid body neurons in the medulla terminalis evoked postsynaptic potentials (50- to 200-ms duration) with an unidentified target in the medulla terminalis. Transient absorption signals were not detected in either the internal or external medulla nor were they recorded from other parts of the optic lobes in response to electrical stimulation of axons of the deutocerebral projection neurons. Functional maps of optical activity, together with electrophysiological and pharmacological findings, suggest that γ-aminobutyric acid affects synaptic transmission in glomeruli of the hemiellipsoid body. Synapses of the olfactory pathway located in the medulla terminalis may act as a “filter,” modifying visual information processing during olfactory stimulation.

Sign in / Sign up

Export Citation Format

Share Document