Intracellular patch clamp techniques. Medical search. Frequent questions

Cultured cells can be studied by means of two fundamental recording techniques that are intracellular and extracellular recordings. (tudelft.nl)
While these techniques result in larger signals which are easier to measure, intracellular or patch-clamp techniques are very challenging in live animals, making extracellular recordings the technique of choice for many investigators. (hindawi.com)
Electrophysiological measurements using the whole cell version of the patch-clamp technique were made simultaneously with [Ca 2+ ](i) microfluorimetric recordings. (umn.edu)
In in vitro slices prepared from rat sensorimotor cortex, intracellular recordings were obtained from 107 layer V pyramidal neurons, subsequently injected with biocytin for morphological reconstruction. (unimib.it)
Extracellular and intracellular recordings in vivo. (weizmann.ac.il)
DSI) was investigated using fura-2 Ca2+ measurements and whole-cell patch-clamp recordings in rat cerebellar Purkinje cells. (ox.ac.uk)
In summary, whole-cell patch-clamp recordings in brain slices provide means to measure in ex vivo preparation long-lasting changes in neuronal functions that have developed in intact awake animals. (elsevierpure.com)
Electrophysiology Patch\clamp recordings had been performed in entire\cell construction at room temp using Axonpatch 200B (Axon, USA) or HEKA EPC10 (HEKA, Germany) amplifier. (holyexperiment.org)
The developed technique, which yields highly parallelized electroporations by using stimulation voltages around 1 Volt peak-to-peak amplitude, enabled intracellular-like recordings at high success rates without causing significant alteration in key electrophysiological features. (mxwbio.com)
Using whole-cell patch-clamp recordings on brain slices of 21- to 25-day-old rats, the present study investigated the effects of GHB and ethanol on spontaneous excitatory postsynaptic currents (sEPSCs) in dopaminergic neurons of the substantia nigra pars compacta (SNc). (123dok.org)
To test these effects, whole cell patch clamp recordings were conducted on acute rat lumbar spinal cord slices. (silverchair.com)
Through paired patch-clamp recordings, we explore how two simultaneously recorded neurons (whether of the same or different cell types) are interconnected and how they influence the electrical properties of each other. (solteszlab.com)

Also, characteristics of the Cl- currents induced by dopamine were observed in whole-cell patch-clamp recording. (nih.gov)
Exogenous ascorbate also induces a moderate increase in programmed cell death symptoms in intact roots, but it does not activate Ca2+ influx currents in patch-clamped root protoplasts. (deepdyve.com)
We recently reported that the time course of these synaptic currents is slower when intracellular chloride is high. (ox.ac.uk)
Here we extend these findings to measure the effects of both extracellular and intracellular chloride on the deactivation of glycine and GABA currents at both negative and positive holding potentials. (ox.ac.uk)
Currents were elicited by fast agonist application to outside-out patches from HEK-293 cells expressing rat glycine or GABA receptors. (ox.ac.uk)
The action of thapsigargin on intracellular calcium homeostasis and voltage-activated calcium currents was studied on freshly isolated adult mouse dorsal root ganglia neurons. (uaeu.ac.ae)
transmembrane Ca 2+ currents were recorded under voltage-clamp in the whole-cell configuration of the patch-clamp technique. (uaeu.ac.ae)
We determined the effects of HMGB1 on isolated feline cardiac myocytes by measuring sarcomere shortening in contracting cardiac myocytes, intracellular Ca 2+ transients by using fluo-3, and L-type calcium currents by using whole cell perforate configuration of the patch-clamp technique. (houstonmethodist.org)
A significant inhibition of the inward L-type calcium currents was also documented by the patch-clamp technique. (houstonmethodist.org)
mu-Opioid agonists have no effect on calcium currents (I(Ca)) in neurohypophysial terminals when recorded using the classic whole-cell patch-clamp configuration. (umassmed.edu)

Intracellular Ca 2+ stores were studied in sympathetic neurons grown in primary culture from the superior cervical ganglion of the rat. (umn.edu)
We conclude that mammalian sympathetic neurons possess intracellular Ca 2+ stores with pharmacological characteristics that closely resemble those found in muscle but that these are relatively small and produce little amplification of [Ca 2+ ](i) transients resulting from Ca 2+ influx through voltage-sensitive Ca 2+ channels. (umn.edu)
Thayer, SA , Hirning, LD & Miller, RJ 1988, ' The role of caffeine-sensitive calcium stores in the regulation of the intracellular free calcium concentration in rat sympathetic neurons in vitro ', Molecular Pharmacology , vol. 34, no. 5, pp. 664-673. (umn.edu)
Patch clamp recording in cultured neurons. (weizmann.ac.il)
However, 5-10 min incubation of neurons with 20 nM thapsigargin completely and almost irreversibly inhibited caffeine-mediated Ca 2+ release from intracellular pools. (uaeu.ac.ae)
Oligomeric and monomeric αSyn were injected directly into the soma of pyramidal neurons in mouse neocortical brain slices during whole-cell patch clamp recording. (nih.gov)
We also employ optogenetic techniques to alter the activity of genetically targeted neuronal populations in response to light stimulation, both to examine their downstream connectivity and how they influence the electrical properties of other neurons. (solteszlab.com)
Neurons recorded with whole cell electrophysiology can be filled with intracellular tracers, allowing enabling later morphological reconstruction of the cell. (solteszlab.com)

Consequently, within this framework, the phase signal will be predominantly sensitive to any mechanism modifying the concentration of intracellular compounds, including transmembrane water movements, which accompany various ionic fluxes. (jneurosci.org)
The G297 residue is located in the intracellular linker of transmembrane segments S2 and S3, and is thus not involved in direct binding of Mg(2+) ions. (fsu.edu)
We have recently introduced a novel technique that combines total internal reflection fluorescence (TIRF) microscopy and whole-cell patch clamp to visualize Ca2+ influx as fluorescence puffs, control the membrane potential, and measure the transmembrane current. (inscreva-se.net)

Ratiometric Ca2+ measurements indicated that the extent of DSI depended critically on the changes in intracellular calcium ([Ca2+]i). (ox.ac.uk)
In this configuration, the micropipette is in tight contact with the cell membrane, which prevents current leakage and thereby provides more accurate ionic current measurements than the previously used intracellular sharp electrode recording method. (elsevierpure.com)
In this study, function and expression of TRPV4 channels in rat carotid artery (CA) were characterized by employing in-situ patch-clamp techniques and single-cell-RT-PCR analysis, measurements of Ca2+-influx through TRPV4, and conducted pressure-myograph experiments in CA and small-sized A. gracilis to test vasodilatory responses after pharmacological activation of the channel by 4αPDD and after increasing wall shear stress (WSS). (tu-berlin.de)

Ca2+](i)-dependent modulation of the inhibitory effect of SEA0400 on NCX was characterized by the changes in Ni2+-sensitive current in voltage-clamped myocytes. (unideb.hu)

This inhibition was associated with deceleration of the recovery of depolarization-induced [Ca 2+ ] i transients, presumably due to the inhibition of Ca 2+ uptake by intracellular calcium stores. (uaeu.ac.ae)
Methods and results Intracellular Ca2+ ([Ca2+](i)) transients, induced by either field stimulation or caffeine flush, were monitored using Ca2+ indicator dyes. (unideb.hu)

Most TRP channels are composed of 6 membrane-spanning helices with intracellular N- and C-termini. (wikipedia.org)
These observations suggest that permeant chloride affects gating by a foot-in-the-door effect, binding to a channel site with asymmetrical access from the intracellular and extracellular sides of the membrane. (ox.ac.uk)
Additionally, significant opening of the plasma membrane allows the internal pipette solution to freely diffuse into the cytoplasm, providing means for introducing drugs, e.g., agonists or antagonists of specific intracellular proteins, and manipulating these targets without altering their functions in neighboring cells. (elsevierpure.com)
The activation of CatSper by progesterone and prostaglandins, as well as the ligand-independent regulation of the channel by a change in the membrane voltage and intracellular pH are going to be addressed. (123dok.net)
Whole-cell patch clamp technique briefly applies large suction to a part of the cell membrane causing disruption and a closed gigaohm seal to allow the cytosol to be one sphere with the intracellular buffer chamber. (reactionbiology.com)
The whole cell patch clamp technique was used to monitor membrane capacitance (Cm) and to load both calcium indicator dye and photolabile chelators into rat pituitary melanotrophs prior to flash photolysis. (mpg.de)
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. (yale.edu)

The retrograde inhibition of IPSCs in rat cerebellar purkinje cells is highly sensitive to intracellular Ca2+. (ox.ac.uk)
However, mu-opioid receptor (MOR)-mediated inhibition of I(Ca) is reliably demonstrated using the perforated-patch configuration. (umassmed.edu)

Gall D, Prestori F, Sola E, D'Errico A, Roussel C, Forti L, Rossi P, D'Angelo E (2005) Intracellular calcium regulation by burst discharge determines bidirectional long-term synaptic plasticity at the cerebellum input stage. (yale.edu)

Our results show a rise in basal intracellular calcium ([Ca(2+)]i) in response to application of [D-Ala(2)-N-Me-Phe(4),Gly5-ol]-Enkephalin (DAMGO), a MOR agonist, that is blocked by D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP), a MOR antagonist. (umassmed.edu)
Activation of TRPV4 by 4αPDD increased intracellular [Ca2+]i significantly above basal levels. (tu-berlin.de)

In the present work, therefore, the concentration-dependent effects of SEA0400 and KB-R7943 on INa/Ca and ICa were studied and compared in canine ventricular cardiomyocytes using the whole-cell configuration of the patch clamp technique. (unideb.hu)
Insulin-induced β-cell hyperpolarization was sufficient to abolish intracellular calcium concentration ([Ca 2+ ] i ) oscillations measured in pancreatic islets exposed to 10 mmol/l glucose. (diabetesjournals.org)

In-vitro cell culturing technique is widely used in many branches of science such as in neuroscience and cardiology to investigate the dynamics of large population of cells. (tudelft.nl)
With his highly unusual scientific background in neuroscience, he has introduced novel techniques aiming at early diagnosis and non-toxic therapy of solid cancers. (sophion.co.jp)

Intriguingly, the replacement of gluconate with ascorbate in the patch-clamp pipette reveals a large ascorbate efflux current, which shows sensitivity to the anion channel blocker, anthracene-9-carboxylic acid (A9C), indicative of the ascorbate release via anion channels. (deepdyve.com)
was observed immediately after continuity between the patch pipette and the cell cytosol was achieved. (mpg.de)

This hypothesis is supported by the observation that the sensitivity of channel gating to intracellular chloride is abolished if the channel is engineered to become cation selective or if positive charges in the external pore vestibule are eliminated by mutagenesis. (ox.ac.uk)
A mutant version of the Cavβ subunit lacking the first forty-six amino acids, including a string of twenty-two acidic residues, no longer conferred sensitivity to intracellular Mg2+-ATP and Na+ ions, while continuing to show wild type modulation of current amplitude and inactivation of Cav2.3. (mblwhoilibrary.org)

Using intracellular Ca 2+ imaging, patch clamp recording and molecular docking techniques, we assessed the function of N338S mutant heterologously expressed in HEK293 cells and HL-1 cardiac cells. (elsevierpure.com)

Voltage dependence is abolished by the same mutations that suppress the effect of intracellular chloride and also by replacing chloride with another permeant ion, thiocyanate. (ox.ac.uk)
Both voltage-gated and ligand-gated channels can be tested using manual patch clamping. (reactionbiology.com)
Using the perforated patch-clamp technique and ratio-calcium-imaging methods, we describe a diffusible second messenger pathway stimulated by the MOR that inhibits voltage-gated calcium channels in isolated terminals from the rat neurohypophysis (NH). (umassmed.edu)

This observation is of particular interest since it shows that DHM is the first imaging technique able to monitor dynamically and in situ the activity of these cotransporters during physiological and/or pathological neuronal conditions. (jneurosci.org)
A major advantage of this technique is that it provides information on how specific manipulations (e.g., pharmacological, experimenter-induced plasticity) may alter specific neuronal functions or channels in real-time. (elsevierpure.com)

The lecture-based part of the course will cover a range of methods, including neurophysiology, imaging, microscopy and optogenetic techniques. (weizmann.ac.il)
Digital holographic microscopy (DHM) is a noninvasive optical imaging technique that provides quantitative phase images of living cells. (jneurosci.org)

Whole-cell patch-clamp recording is an electrophysiological technique that allows the study of the electrical properties of a substantial part of the neuron. (elsevierpure.com)

We found that expression of TRPC6 N338S significantly increased intracellular Ca 2+ levels ([Ca 2+ ] i ) and current densities in response to 50 μM 1-oleoyl 2-acetyl-sn-glycerol (OAG), an activator of TRPC6 channels, compared to those of TRPC6 WT. (elsevierpure.com)

Cohen D, Yarom Y (1998) Patches of synchronized activity in the cerebellar cortex evoked by mossy-fiber stimulation: questioning the role of parallel fibers. (yale.edu)

The increases of intracellular cAMP and Ca2+ induced by dopamine were also measured. (nih.gov)

Caffeine (10 mM) and ryanodine (10 μM), applied singly or together, produced a small and variable decrease in the [Ca 2+ ](i) transient resulting from cell depolarization using the whole-cell patch-clamp technique. (umn.edu)
Here, we focus on the modulatory phenotype of the conventional Cavβ subunit (SmCavβ) using the human Cav2.3 channel as the substrate for SmCavβ and the whole-cell patch-clamp technique. (mblwhoilibrary.org)

Here, we demonstrate how porous Pt-black electrodes, arranged in high-density microelectrode arrays, can be used to record intracellular-like signals of cardiomyocytes at large-scale repeatedly over an extended period of time. (mxwbio.com)

Together, these results suggest an important role of the intracellular linker regions in gating processes of Eag channels. (fsu.edu)

In particular, when combined with appropriate pharmacology, this technique is a powerful tool allowing identification of specific neuroadaptations that occurred following any type of experiences, such as learning, exposure to drugs of abuse, and stress. (elsevierpure.com)

Read and understand research papers that utilize modern molecular approaches, neurophysiology and imaging techniques. (weizmann.ac.il)

In addition to confirming the activity of potential hits from high or medium-throughput screens, manual patch clamping can be used to assess the mechanism of action of compounds and to determine the effects of compounds on the biophysical properties of a channel. (reactionbiology.com)

This technique has been particularly useful to assess function and dysfunction of the oropharyngeal phase of swallowing. (nature.com)

Superfusion of the cells with 10 mM caffeine elicited a rapid and transient increase in [Ca 2+ ](i) in the absence of extracellular Ca 2+ , indicating the presence of a caffeine-sensitive intracellular Ca 2+ storage site. (umn.edu)
In summary, this technique has immensely contributed to the understanding of passive and active biophysical properties of excitable cells. (elsevierpure.com)
Our fully automated patch clamp platform comprises QPatch HTX and QPatch 16 that allows for the testing of up to 48 or 16 cells in parallel, respectively. (reactionbiology.com)
Using the perforated patch clamp technique, we found that insulin (1-600 nmol/l) but not IGF-1 (100 nmol/l) reversibly hyperpolarized single mouse β-cells and inhibited their electrical activity. (diabetesjournals.org)

Research Fellow, Laboratory of Experimental Neurosurgery, Institut de Recherche Interdisciplinaire en Biologie Humaine et Moléculaire. (silverchair.com)

To investigate proarrhythmic effects of drugs, the patch clamp technique has been used as the gold standard for charac- terizing the electrophysiology of cardiomyocytes in vitro. (mxwbio.com)

Elimination of Ca2+ from the extracellular solution prevented any fluorescence increase after the application of either milli- or nanosecond pulses, indicating that intracellular Ca2+ stores were not involved in puff formation. (inscreva-se.net)

The importance of diverse intracellular organellar ion channels in a wide variety of diseases is now gaining greater attention (e.g. (metrionbiosciences.com)
In cell-attached patches, single ATP-sensitive K + (K ATP ) channels were activated by bath-applied insulin and subsequently inhibited by wortmannin. (diabetesjournals.org)

The slowing effect of high extracellular chloride on current decay was detectable only in low intracellular chloride (4 mm). (ox.ac.uk)
For selectivity evaluation, test compound (0.1?mm) was added to the intracellular answer (ICS) to determine intracellular effect of compound on individual TRPM7, TRPM8, TRPV1 and TRPV3, respectively. (holyexperiment.org)
This suppressive effect was independent of Ca2+, but dependent on intracellular Mg2+-ATP. (mblwhoilibrary.org)

The main advantages of this technique are the prompt integrability with any electrophysiologist type, the potentially high special and temporal resolution, they reduce the invasivity. (jove.com)

Manual patch clamping is the "gold-standard" for the investigation of ion channel activity. (reactionbiology.com)

Preloading monolayers with the intracellular Ca2+ chelator BAPTA/AM abolished the initial spike and greatly reduced the second phase in the Isc response to dopamine. (nih.gov)
State-of-the-art techniques allow researchers to record large numbers of spike trains in parallel for many hours. (nature.com)

As the obtained results were in good agreement with previously published data, we are confident that the devel- oped technique has the potential to be routinely used in in vitro platforms for cardiotoxicity screening. (mxwbio.com)
Moreover, mutation of the only positively charged residue in the other intracellular linker between S4 and S5 also results in a dramatic reduction of Mg(2+)-dependent modulation of Eag activation kinetics. (fsu.edu)

This 'loading transient' reflects the release of calcium from DMN during the binding of intracellular magnesium. (mpg.de)

This method was modified to analyze pore formation by applying electric pulses extracellularly, without patch clamp. (inscreva-se.net)

The main emphasis will be given on techniques of neurophysiology. (weizmann.ac.il)

A Feature Paper should be a substantial original Article that involves several techniques or approaches, provides an outlook for future research directions and describes possible research applications. (mdpi.com)

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