Journal ArticleDOI
Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.
TLDR
The extracellular patch clamp method, which first allowed the detection of single channel currents in biological membranes, has been further refined to enable higher current resolution, direct membrane patch potential control, and physical isolation of membrane patches.Abstract:
1. The extracellular patch clamp method, which first allowed the detection of single channel currents in biological membranes, has been further refined to enable higher current resolution, direct membrane patch potential control, and physical isolation of membrane patches. 2. A description of a convenient method for the fabrication of patch recording pipettes is given together with procedures followed to achieve giga-seals i.e. pipette-membrane seals with resistances of 10(9) - 10(11) omega. 3. The basic patch clamp recording circuit, and designs for improved frequency response are described along with the present limitations in recording the currents from single channels. 4. Procedures for preparation and recording from three representative cell types are given. Some properties of single acetylcholine-activated channels in muscle membrane are described to illustrate the improved current and time resolution achieved with giga-seals. 5. A description is given of the various ways that patches of membrane can be physically isolated from cells. This isolation enables the recording of single channel currents with well-defined solutions on both sides of the membrane. Two types of isolated cell-free patch configurations can be formed: an inside-out patch with its cytoplasmic membrane face exposed to the bath solution, and an outside-out patch with its extracellular membrane face exposed to the bath solution. 6. The application of the method for the recording of ionic currents and internal dialysis of small cells is considered. Single channel resolution can be achieved when recording from whole cells, if the cell diameter is small (less than 20 micrometer). 7. The wide range of cell types amenable to giga-seal formation is discussed.read more
Citations
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Journal ArticleDOI
Synaptic Modifications in Cultured Hippocampal Neurons: Dependence on Spike Timing, Synaptic Strength, and Postsynaptic Cell Type
Guo-Qiang Bi,Mu-ming Poo +1 more
TL;DR: The results underscore the importance of precise spike timing, synaptic strength, and postsynaptic cell type in the activity-induced modification of central synapses and suggest that Hebb’s rule may need to incorporate a quantitative consideration of spike timing that reflects the narrow and asymmetric window for the induction of synaptic modification.
Journal ArticleDOI
Magnesium gates glutamate-activated channels in mouse central neurones
TL;DR: The voltage dependence of the NMDA receptor-linked conductance appears to be a consequence of the voltage dependenceof the Mg2+ block and its interpretation does not require the implication of an intramembrane voltage-dependent ‘gate’.
Journal ArticleDOI
Glycine potentiates the NMDA response in cultured mouse brain neurons
J. W. Johnson,P Ascher +1 more
TL;DR: G glycine may facilitate excitatory transmission in the brain through an allosteric activation of the NMDA receptor, and can be observed in outside-out patches as an increase in the frequency of opening of the channels activated by NMDA agonists.
Journal ArticleDOI
Heteromeric NMDA receptors: Molecular and functional distinction of subtypes
Hannah Monyer,Rolf Sprengel,Ralf Schoepfer,Anne Herb,Miyoko Higuchi,Hilda Lomelí,Nail Burnashev,Bert Sakmann,Peter H. Seeburg +8 more
TL;DR: Molecular cloning identified three complementary DNA species of rat brain, encoding NMDA receptor subunits NMDAR2A (NR2A), NR2B, and NR2C, which are 55 to 70% ientical in sequence, and these are structurally related, with less than 20% sequence identity, to other excitatory amino acid receptor sub Units.
Journal ArticleDOI
Channelrhodopsin-2, a directly light-gated cation-selective membrane channel.
Georg Nagel,Tanjef Szellas,Wolfram Huhn,Suneel Kateriya,Nona Adeishvili,Peter Berthold,Doris Ollig,Peter Hegemann,Ernst Bamberg +8 more
TL;DR: It is demonstrated by functional expression, both in oocytes of Xenopus laevis and mammalian cells, that ChR2 is a directly light-switched cation-selective ion channel, and may be used to depolarize small or large cells, simply by illumination.
References
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Journal ArticleDOI
Single-channel currents recorded from membrane of denervated frog muscle fibres
Erwin Neher,Bert Sakmann +1 more
TL;DR: A more sensitive method of conductance measurement is reported, which, in appropriate conditions, reveals discrete changes in conductance that show many of the features that have been postulated for single ionic channels.
Journal ArticleDOI
Single acetylcholine-activated channels show burst-kinetics in presence of desensitizing concentrations of agonist
TL;DR: From the temporal sequence of the pulses, derived estimates of the rates of activation and desensitization of the AChR-channel are derived and it is reported that in the presence of ACh concentrations sufficient to cause desensItization, single channel current pulses appear in groups.
Journal ArticleDOI
The extracellular patch clamp: a method for resolving currents through individual open channels in biological membranes
TL;DR: The current contributions of individual ionic channels can be measured by electrically isolating a small patch of membrane by bringing the tip of a small pipette into close contact with an enzymatically cleaned membrane of a hypersensitive amphibian or mammalian muscle fiber.
Journal ArticleDOI
A receptor for protons in the nerve cell membrane
TL;DR: It is suggested that this newly described conductance mechanism may serve as a pH-sensor in the sensory nerve endings throughout the body.
Journal ArticleDOI
Measured work of deformation and repulsion of lecithin bilayers
TL;DR: A first determination of the lateral pressure required to create large increases in bilayers thickness and concomitant decreases in bilayer area is obtained, enabling critical tests of theories on bilayer stability and phase transition.
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