Patch-clamp technique to characterize ion channels in enlarged individual endolysosomes

2017 ◽  
Vol 12 (8) ◽  
pp. 1639-1658 ◽  
Author(s):  
Cheng-Chang Chen ◽  
Chunlei Cang ◽  
Stefanie Fenske ◽  
Elisabeth Butz ◽  
Yu-Kai Chao ◽  
...  
Keyword(s):  
Ion Channels ◽  
Patch Clamp ◽  
Patch Clamp Technique ◽  
Clamp Technique

Patch-clamp technique in renal physiology

1986 ◽  
Vol 250 (3) ◽  
pp. F379-F385 ◽  
Author(s):  
L. G. Palmer
Keyword(s):  
Ion Channels ◽  
Patch Clamp ◽  
Ion Transport ◽  
Renal Physiology ◽  
Patch Clamp Technique ◽  
Clamp Technique ◽  
Basic Concepts

The patch-clamp technique of Neher and Sakmann and their colleagues has been widely used over the last 5 years to investigate ion channels in excitable tissues. More recently, it has become useful as a tool to study channels involved in transepithelial ion transport. In this review, I briefly cover the basic concepts behind the patch-clamp technique and the kinds of information that can be obtained with it. I then summarize the applications of the technique to renal tissues and describe some of the channel types that have been observed to date in epithelia.

Electrophysiological investigation of anthelmintic resistance

Parasitology ◽  
2000 ◽  
Vol 120 (7) ◽  
pp. 87-94 ◽  
Author(s):  
R. J. MARTIN ◽  
A. P. ROBERTSON
Keyword(s):  
Ion Channels ◽  
Patch Clamp ◽  
Anthelmintic Resistance ◽  
Resistant Isolate ◽  
Patch Clamp Technique ◽  
Clamp Technique ◽  
Receptor Sites ◽  
Flow Through ◽  
Sites Of Action ◽  
A Site

It is pointed out that two of the three major groups of anthelmintic act by opening membrane ion-channels. It is appropriate, therefore, to use electrophysiological methods to study the properties of the sites of action of these drugs and the changes in the properties of these receptor sites associated with resistance. This paper describes the use of the patch- clamp technique to observe the currents that flow through the levamisole-activated channels as they open and close in levamisole-sensitive and levamisole-resistant isolates. It was found that, on average, the proportion of time the channels are open, is less in the resistant isolate. The patch-clamp technique also showed that the ion-channels are heterogeneous and that one of the subtypes is lost with the appearance of resistance. The use of the current clamp technique is illustrated to record a site of action of ivermectin in the pharyngeal muscle of Ascaris.

Patch-Clamp Recording of Ion Channels: Interfering Effects of Patch Pipette Glass

Physiology ◽  
1990 ◽  
Vol 5 (4) ◽  
pp. 155-158
Author(s):  
C Zuazaga ◽  
A Steinacker
Keyword(s):  
Ion Channels ◽  
Patch Clamp ◽  
Ion Channel ◽  
Channel Activity ◽  
Patch Clamp Recording ◽  
Patch Clamp Technique ◽  
Clamp Technique ◽  
Ion Channel Properties ◽  
Patch Pipette ◽  
Channel Properties

Glass for pipettes used to record ion channel activity with the patch-clamp technique is selected on the basis of its electrical, thermal, and sealing properties. Recent findings stress a new characteristic to consider: the effect of pipette glass itself on ion channel properties.

scholarly journals Patch-clamp technique for studying ion channels in activated platelets

2021 ◽  
Vol 1 (1) ◽  
pp. 3-12
Author(s):  
Anatoli Kokhan ◽  
Semyon Zdanevich ◽  
Igor Prokofev ◽  
Irina Gorudko ◽  
Ekaterina Shamova
Keyword(s):  
Ion Channels ◽  
Patch Clamp ◽  
Patch Clamp Technique ◽  
Clamp Technique ◽  
Activated Platelets

Shock Wave Therapy as an Innovative Technology in Skeletal Disorders: Study on Transmembrane Current in Stimulated Osteoblast-Like Cells

2005 ◽  
Vol 28 (8) ◽  
pp. 841-847 ◽  
Author(s):  
L. Martini ◽  
G. Giavaresi ◽  
M. Fini ◽  
P. Torricelli ◽  
V. Borsari ◽  
...  
Keyword(s):  
Shock Wave ◽  
Ion Channels ◽  
Patch Clamp ◽  
Ionic Current ◽  
Innovative Technology ◽  
Patch Clamp Recording ◽  
Patch Clamp Technique ◽  
Transmembrane Current ◽  
Clamp Technique ◽  
And Control

Extracorporeal shock wave treatment (ESWT) is successfully used in various musculoskeletal disorders and pathologies. Despite the increasing use of this kind of therapy, some aspects of its mechanism of action are still unclear. In vitro bone cell behavior under ESWT were previously investigated by the present author and MG63 osteoblast-like cells showed an enhancement in proliferation and in the osteoblast differentiation after therapy with a low-energy flux density. The aim of the present study was to evaluate the effect of ESWT on the permeabilization of cell membrane. We characterized physiological changes in the MG63 associated with ESWT generated by an ESW device and patch clamp recording was performed to study ion channels. Experiments were carried out using the whole-cell recording configuration of the patch-clamp technique and the ionic current measurements were performed on cell samples of ESW treated and control groups. The patch-clamp technique showed the effect of ESWT on the amplitude of transmembrane currents. The treatment with ESW enhanced the transmembrane current as well the voltage dependence of Ca-activated and K channels that mediate these currents: the differences between treated cells and control at 80mV were over 1000 pA (P<0.05). These modifications of ion channels activity positively influence cell proliferation (MTT test, P<0.0001) without interfering with the normal synthesis activity of stimulated osteoblasts.

scholarly journals Electrophysiology study using patch clamp technique in sensory neurons

2021 ◽  
Vol 1 (1) ◽  
pp. 12-14
Author(s):  
Tiago Dos Santos-Nascimento ◽  
Kleyane Morais Veras ◽  
José Ossian Almeida Souza-Filho ◽  
Luiz Moreira-Júnior
Keyword(s):  
Plasma Membrane ◽  
Ion Channels ◽  
Patch Clamp ◽  
Pharmacological Study ◽  
Ionic Currents ◽  
Outer Diameter ◽  
Patch Clamp Technique ◽  
Pharmacological Agents ◽  
Clamp Technique ◽  
Tip Resistance

The electrophysiological and pharmacological study involving sensory and autonomic neurons enables the development of new effective agents in the treatment of neuropathic disorders, since they enable the elucidation of the mechanisms underlying the malfunction of the nervous system. In this context, the patch clamp technique increased the study of cells, providing a high-resolution method at the molecular level for observing the flow of ions through ion channels characteristic of excitable cells [1], such as the neurons. When using different protocols with combinations of intracellular and extracellular solutions with specific pharmacological agents, this technique allows different unit and/or macroscopic records of active and passive electrical variables of cellular activity [2] that it favored the Nobel Prize in physiology or medicine to Erwin Neher and Bert Sakmann in 1991. Although the whole cell mode is the most used configuration in health-related researches, little is known in health courses. To apply this technique to neurons, it is commonly necessary to dissociate neurossomas. Figure 01 shows sensory neurossome of the dorsal root ganglion (GRD) of rats from the bioterium of the State University of Ceará (CEUA process number 10339956-9). The process of isolating neurossomas from the intact ganglion consists of two phases: 1) Collagenase (1mg / ml for 75 min) and Trypsin + EDTA (0.25% and 0.025%, respectively, for 12 minutes); 2) Mechanical dispersion with 3 Pasteur glass pipettes with decreasing diameter (2.5 mm, 1 mm and 0.5 mm, respectively). Then, the neurossomas were plated on coverslips previously treated with poly-D-lysine maintained in supplemented DMEM and incubated at 37 °C and 5% CO2 [3]. The figure shows a neurossoma 24h after plating. This cell has approximately 25 µM in diameter, which it plays role nociception function [4]. Furthermore, the nucleus is not centralized, the cell does not have neurites. As for the micropipette, capillaries were used for micro-hematocrit without heparin (75 mm length, 1 mm inner diameter and 1.5 mm outer diameter) for making with tip resistance range from 1 and 3 MΩ after filling with the solution to compose intracellular medium [5]. In this technique, a microelectrode was micrometrically move toward until it lightly touched the plasma membrane. Then, a continuous negative pressure was applied to increase the contact of the glass with the membrane, stabilizing the seal (interaction between membrane and glass) and increasing it until its resistance reaches the order of 109 ohm (GΩ). Then, more suction was applied to cause the cell surface under the microelectrode to rupture, thus providing access to the interior of the cell, allowing excellent control of the cell membrane potential and, consequently, high-fidelity records of ionic currents that flow through ion channels present in the plasma membrane of neurossomas.

The Patch-Clamp Technique: Recording Ionic Currents Through Single Pores in the Cell Membrane

Physiology ◽  
1986 ◽  
Vol 1 (1) ◽  
pp. 5-8
Author(s):  
OH Petersen ◽  
CCH Petersen
Keyword(s):  
Ion Channels ◽  
Cell Membrane ◽  
Patch Clamp ◽  
Ionic Currents ◽  
State Model ◽  
Patch Clamp Technique ◽  
Clamp Technique ◽  
Individual Ion ◽  
Two State Model

When an intracellular electrode records an electrical event in the cell membrane, we know that there are changes in the membrane's permeability to various ions. These changes are mediated by membrane entities known as pores or ion channels. The advance of the patch-clamp technique, which permits the study of changes in individual ion channels, in the simplest case conforms to a simple open-close two-state model.

Sign in / Sign up

Export Citation Format

Share Document