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Until the mid-1960s, studies on the metabolism of brain mitochondria were hampered by the lack of suitable methodologies for brain mitochondrial isolation. In the majority of the studies up to this time, crude mitochondrial preparations were isolated from mammalian brain homogenates ...

The synapse plays pivotal roles in various neuronal functions, with its efficacy determining the opening and closing of neuronal circuits. Among the parameters that determine synaptic efficacy, presynaptic factors are of crucial importance. However, these factors are among those ...

Since late 1980s, the whole-cell patch-clamp technique has been used as a powerful tool for analyzing local circuits of the central nervous system in a brain slice preparation in which the fundamental architecture of local circuits is mostly maintained. When combined with intracellular s ...

The practical details of the “blind” patch-clamp recording method and its usefulness and applications to the analysis of synaptic transmission and plasticity in brain slice preparations are described. The blind patch-clamp method displays its greatest power in the experiments in wh ...

Patch-clamp is the best method for analyzing single-channel current activities today. The method is not good, however, at measuring the total number of channels expressed on the cell membrane that are responsible for the current of concern. In contrast, noise analysis has some advantage over ...

Analysis of data from single-channel studies can provide us with insights into the detailed physicochemical features of channel proteins, such as ion permeation rate, ion selectivity, and gating. However, single-channel analysis, particularly gating analysis, is sometimes time ...

The patch-clamp technique has revolutionized the study of membrane physiology, enabling unprecedented resolution in recording cellular electrical responses and underlying mechanisms. The perforated-patch variant of whole-cell patch-clamp recording was developed to ...

With the whole-cell mode of patch-clamp the membrane current is recorded from a cell under the voltage-clamp. The membrane potential response of a cell is also measured and manipulated under a current-clamp. The method enables one to correlate the macroscopic aspects of ion channels with their ...

The patch-clamp technique, invented by Erwin Neher and Bert Sakmann in 1976, was originally designed to detect the activity of single-ion channel proteins in the cell membrane. The technique is now widely used in the field of ion channel research for many purposes, such as to monitor changes in the tot ...

Solutions used in patch-clamp can be divided into two general categories by the side of the patch membrane with which the solution is in contact. The compositions of extracellular solutions are usually similar to those of the natural extracellular fluids, whereas intracellular solutio ...

Patch-clamp data, including channel currents and membrane voltage, are usually recorded to a computer via a process known as analog-to-digital conversion. This chapter gives essential information on how to avoid pitfalls and obtain error-free data with optimal resolution.

The physical dimensions of an ion channel pore can be estimated in patch-clamp experiments using various charged and noncharged molecular probes. Measuring the permeability to organic ions of different size and shape yields an estimate of the size of the narrowest part of the selectivity fi ...

When a patch pipette approaches a cellular surface at distances comparable to the size of the tip opening, the monitored pipette current drops before the pipette touches the membrane. Fine nano-tipped glass electrodes can effectively function as “spherical sensors” that can “roll” over s ...

If gene expression could be analyzed after electrophysiological recording of a single cell, it would enhance our molecular understanding of neuronal physiology and help to interpret the results obtained from patch-clamp recordings. For instance, after completing a recording from a ...

This chapter describes practical techniques for electrophysiological experiments in the heterologous expression systems using mammalian cell cultures and Xenopus laevis oocytes. These two experimental systems are the most commonly used for expression of ion channels and t ...

Since the cloning of the first thermosensitive TRP channel, a capsaicin receptor TRPV1, in 1997, nine thermosensitive TRP channels have been identified. Thermosensitive TRP channels can be used to detect local temperatures by expression in HEK293 cells as a kind of biosensor, as described ...

Cells or membranes with ligand-gated ion channels endogenously expressed in cultured cells or heterologously expressed in HEK293 cells can be used as biosensors with a patch-clamp technique to detect small amounts of locally released bioactive substances that are known to activate i ...

Patch-clamping is an essential technique in the field of ion channel drug discovery and safety pharmacology. Conventional patch-clamp methods provide high data quality and can be applied to a wide variety of cells, but the methods are laborious and time-consuming. Automated patch-clamp ...

A historical overview describes how classic electrophysiological techniques, such as membrane potential measurement, seal formation, and the voltage clamp, have evolved into modern patch-clamp techniques. We show that old ideas from seminal papers on ion channels (i.e., gating, in ...

When the second patch-clamp book of Sakmann and Neher appeared in 1995 (Sakmann and Neher, Single-channel recording, 2nd edn. Plenum Press, New York, 1995), the molecular nature of plant ion channels was still in its infancy. Since 1995, various members of the Shaker-, Two-Pore-, and KCO-type potass ...