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 ...

The standard giga-seal patch-clamp techniques opened the way to monitor ion-channel activity in the native organelle membrane in situ. One has to have an organelle preparation, covered with the intact membrane carrying rightly oriented channel proteins and suitable for proper patch- ...

Not only electrical conductance but also electrical capacitance of the cell membrane can be measured by patch-clamp techniques. Exocytotic events can be detected by recording changes in membrane capacitance. The membrane capacitance, which reflects the surface area of the plasma me ...

The planar lipid bilayer method is another way to examine channel molecules functionally at the single-molecule level. In contrast to patch-clamping, channel molecules are isolated from various biological resources and are reconstituted into an artificial membrane that has a defi ...

The piette perfusion technique is a version of patch-clamp techniques and provides a greater intracellular access during electrophysiological recordings. This internal perfusion technique offers the intracellular change of not only ions but also substances with higher mole ...

The giant patch method was first developed in 1989. The major characteristic is the use of a pipette with a large tip diameter, which dramatically broadened the applications of the excised patch. The giant patch method enables (1) the current recording of transporters/channels with slow turn ...

Na+/K+ pump, Na+/Ca2+ exchanger, glutamate transporter and other transporters for dopamine, norepinephrine, GABA, and others, are all electrogenic. These transporters or exchanger transport Na+ ions, with or without transmitters, in or out of the cells, while K+ or Ca2+, sometimes with anion ...

The advent of two-photon microscopy has enabled us to visualize individual neurons in the intact brain. This technique, used in combination with whole-cell patch-clamp recordings, has facilitated targeted intracellular recording from particular neurons of interest. This chap ...

The methods for in vivo blind patch-clamp recording from the spinal cord, brain stem, and cortex neurons are described herein, including the (1) technique for in vivo preparations, (2) recordings of action potential and synaptic potentials under current-clamp conditions and excitatory ...

Dendritic patch-clamp recordings have recently become popular as an electrophysiological technique to investigate the functional properties of ion channels in neuronal dendrites. During the past decade, experimentalists have pioneered recording from dendrites of neur ...

Over a period spanning some 15 yr, we have exploited microinjected oocytes and embryos of Xenopus laevis to clone and characterize the human genes encoding AChE and BuChE and their protein products. Subsequently, we tapped into the enormous versatility of Xenopus to continue into the realm of t ...

A DNA sequence encoding the brain and muscle form of human AChE (AChE-T) was constructed in our laboratory from cloned cDNA and genomic sequences, and tentatively identified by its homology to known ChEs (Soreq et al., 1990). This putative AChE-coding sequence, bearing the 3′ alternative exon E6, w ...

This chapter is meant to serve two functions. First, we present the methods used to produce the experimental results presented in the following chapters. These are designed to assist the reader in understanding the accompanying figures and to define the specific experimental conditions ...

The complete reproductive cycle of Xenopus spans up to 18 mo from the earliest stages of oogenesis to the sexually mature adult. Owing to its popularity as an embryonic model, the long arduous path to adulthood in Xenopus has been thoroughly documented (see Deuchar, 1966, Deuchar, 1975; Nieuwkoop a ...

The rhesus macaque represents a pragmatic animal model for elucidating mechanisms underlying normal and pathological human behaviors. Many of the same techniques that are used in clinical studies can be readily applied to the nonhuman primate studies. These including the use of Actiwa ...

Uncertainty is an important concept in neuroscience: due to its relevance in everyday life, because of theoretical significance for neurocomputational models, and clinical implications. A body of empirical research has tackled fundamental questions about how uncertainty is re ...

Autism spectrum disorders (ASDs) are characterized by impairments in communication, social interaction, and the presence of restricted and repetitive behaviors. Impairments in the capacity for social interaction include deficits in the use of nonverbal behaviors, failure to d ...