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Single channel patch clamp

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https://www.readbyqxmd.com/read/28075010/stimulatory-actions-of-a-novel-thiourea-derivative-on-large-conductance-calcium-activated-potassium-channels
#1
Sheng-Nan Wu, Jyh-Haur Chern, Santai Shen, Hwei-Hisen Chen, Ying-Ting Hsu, Chih-Chin Lee, Ming-Huan Chan, Ming-Chi Lai, Feng-Shiun Shie
AIM: In this study, we examine whether an anti-inflammatory thiourea derivative, compound #326, actions on ion channels. METHODS: The effects of compound #326 on Ca(2+) -activated K(+) channels were evaluated by patch-clamp recordings obtained in cell-attached, inside-out or whole-cell configuration. RESULTS: In pituitary GH3 cells, compound #326 increased the amplitude of Ca(2+) -activated K(+) currents (IK(Ca) ) with an EC50 value of 11...
January 11, 2017: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/28052704/conflicting-effects-by-antibodies-against-connexin36-during-the-action-of-intracellular-cyclic-amp-onto-electrical-synapses-of-retinal-ganglion-cells
#2
Soh Hidaka
Alpha-type retinal ganglion cells (alpha cells) of the same class in mammalian retina are connected by gap junctions. Electrical synapses between alpha cells were examined using combined techniques of dual patch-clamp recordings, intracellular labeling and electron microscopy in the albino rat retina. In simultaneous dual whole-cell recordings from pairs of neighboring alpha cells, bidirectional electrical synapses with symmetrical junction conductance were observed in pairs with cells of the same morphological type...
January 4, 2017: Journal of Integrative Neuroscience
https://www.readbyqxmd.com/read/28039402/high-and-low-conductance-nmda-receptors-are-present-in-layer-4-spiny-stellate-and-layer-2-3-pyramidal-neurons-of-mouse-barrel-cortex
#3
Christian Scheppach
N-Methyl-D-aspartate (NMDA) receptors are ion channels activated by the neurotransmitter glutamate in the mammalian brain and are important in synaptic function and plasticity, but are also found in extrasynaptic locations and influence neuronal excitability. There are different NMDA receptor subtypes which differ in their single-channel conductance. Recently, synaptic plasticity has been studied in the mouse barrel cortex, the primary sensory cortex for input from the animal's whiskers. Pharmacological data imply the presence of low-conductance NMDA receptors in spiny stellate neurons of cortical layer 4, but of high-conductance NMDA receptors in pyramidal neurons of layer 2/3...
December 2016: Physiological Reports
https://www.readbyqxmd.com/read/27989687/the-role-of-protein-kinase-c-in-the-metabolic-regulation-of-the-cardiac-na-channel
#4
Man Liu, Guangbin Shi, Kai-Chien Yang, Lianzhi Gu, Anumantha G Kanthasany, Vellareddy Anantharam, Samuel C Dudley
BACKGROUND: NADH increases in cardiomyopathy, activates protein kinase C (PKC), upregulates mitochondrial reactive oxygen species (mitoROS), and downregulates the cardiac Na(+) channel (Nav1.5). OBJECTIVE: The objective was to determine how NADH signals downregulation of Nav1.5. METHODS: Isolated mouse cardiomyocytes were used for patch clamp recording and to monitor mitoROS with MitoSOX(TM) Red. HEK293 cells were used for transient transfections...
December 15, 2016: Heart Rhythm: the Official Journal of the Heart Rhythm Society
https://www.readbyqxmd.com/read/27899943/effects-of-allitridum-on-the-transient-outward-potassium-current-in-rats-with-heart-failure
#5
Xiao-Jing Zhao, Kun Lin, Yu Zhang, Bin Xu, Li Liu, Yi-Cheng Fu, Xi Chen, Zhong-Qi Cai, Zhi-Juan Wu, Yun Huang, Yang Li
OBJECTIVE: To study the effect of allitridum on the transient outward potassium current (Ito) of ventricular myocytes in heart failure (HF). METHODS: The dual enzymatic method was used to separate single ventricular myocytes from Sprague Dawley rats. Patch-clamping was used to record Ito and analyze the effect of allitridum on the current. RESULTS: The Ito current had a significant decrease in the HF group, compared with the control group...
September 2016: Journal of Geriatric Cardiology: JGC
https://www.readbyqxmd.com/read/27859335/extracellular-protons-enable-activation-of-the-calcium-dependent-chloride-channel-tmem16a-ano1
#6
Silvia Cruz-Rangel, José J De Jesus-Pérez, Iván A Aréchiga-Figueroa, Aldo A Rodríguez-Menchaca, Patricia Pérez-Cornejo, H Criss Hartzell, Jorge Arreola
TMEM16A (ANO1), the pore forming subunit of a Ca(2+) -dependent Cl(-) channel (CaCC), is activated by direct, voltage-dependent, binding of intracellular Ca(2+) . Endogenous CaCCs are regulated by extracellular protons; however, the molecular basis of such regulation remains unidentified. Here, we evaluated the effects of different extracellular proton concentrations ([H(+) ]o ) on mouse TMEM16A expressed in HEK-293 cells using whole-cell and inside-out patch clamp recordings. We found that increasing the [H(+) ]o from 10(-10) to 10(-5...
November 17, 2016: Journal of Physiology
https://www.readbyqxmd.com/read/27856251/local-calcium-signalling-is-mediated-by-mechanosensitive-ion-channels-in-mesenchymal-stem-cells
#7
Vladislav I Chubinskiy-Nadezhdin, Valeria Y Vasileva, Natalia A Pugovkina, Irina O Vassilieva, Elena A Morachevskaya, Nikolay N Nikolsky, Yuri A Negulyaev
Mechanical forces are implicated in key physiological processes in stem cells, including proliferation, differentiation and lineage switching. To date, there is an evident lack of understanding of how external mechanical cues are coupled with calcium signalling in stem cells. Mechanical reactions are of particular interest in adult mesenchymal stem cells because of their promising potential for use in tissue remodelling and clinical therapy. Here, single channel patch-clamp technique was employed to search for cation channels involved in mechanosensitivity in mesenchymal endometrial-derived stem cells (hMESCs)...
January 22, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27827370/ethanol-sensitive-pacemaker-neurons-in-the-mouse-external-globus-pallidus
#8
Karina P Abrahao, Jessica H Chancey, C Savio Chan, David M Lovinger
Although ethanol is one of the most widely used drugs, we still lack a full understanding of which neuronal subtypes are affected by this drug. Pacemaker neurons exert powerful control over brain circuit function, but little is known about ethanol effects on these types of neurons. Neurons in the external globus pallidus (GPe) generate pacemaker activity that controls basal ganglia, circuitry associated with habitual and compulsive drug use. We performed patch-clamp recordings from GPe neurons and found that bath application of ethanol dose-dependently decreased the firing rate of low-frequency GPe neurons, but did not alter the firing of high-frequency neurons...
December 14, 2016: Neuropsychopharmacology: Official Publication of the American College of Neuropsychopharmacology
https://www.readbyqxmd.com/read/27823931/two-p2x1-receptor-transcripts-able-to-form-functional-channels-are-present-in-most-human-monocytes
#9
Cintya López-López, Josue Jaramillo-Polanco, Diana P Portales-Pérez, Karen S Gómez-Coronado, Jessica G Rodríguez-Meléndez, Juan D Cortés-García, Rosa Espinosa-Luna, Luis M Montaño, Carlos Barajas-López
To characterize the presence and general properties of P2X1 receptors in single human monocytes we used RT-PCR, flow cytometry, and the patch-clamp and the two-electrode voltage-clamp techniques. Most human monocytes expressed the canonical P2X1 (90%) and its splicing variant P2X1del (88%) mRNAs. P2X1 receptor immunoreactivity was also observed in 70% of these cells. Currents mediated by P2X1 (EC50=1.9±0.8µm) and P2X1del (EC50 >1000µm) channels, expressed in Xenopus leavis oocytes, have different ATP sensitivity and kinetics...
December 15, 2016: European Journal of Pharmacology
https://www.readbyqxmd.com/read/27805790/interplay-among-distinct-ca-2-conductances-drives-ca-2-sparks-spontaneous-transient-outward-currents-in-rat-cerebral-arteries
#10
Ahmed M Hashad, Neil Mazumdar, Monica Romero, Anders Nygren, Kamran Bigdely-Shamloo, Osama F Harraz, Jose L Puglisi, Edward J Vigmond, Sean M Wilson, Donald G Welsh
KEY POINTS: Distinct Ca(2+) channels work in a coordinated manner to grade Ca(2+) spark/spontaneous transient outward currents (STOCs) in rat cerebral arteries. The relative contribution of each Ca(2+) channel to Ca(2+) spark/STOC production depends upon their biophysical properties and the resting membrane potential of smooth muscle. Na(+) /Ca(2+) exchanger, but not TRP channels, can also facilitate STOC production. ABSTRACT: Ca(2+) sparks are generated in a voltage-dependent manner to initiate spontaneous transient outward currents (STOCs), events that moderate arterial constriction...
November 2, 2016: Journal of Physiology
https://www.readbyqxmd.com/read/27802315/exosomal-gapdh-from-proximal-tubule-cells-regulate-enac-activity
#11
Kishore Kumar Jella, Ling Yu, Qiang Yue, Daniel Friedman, Billie J Duke, Abdel A Alli
Exosomes are nanometer-scale, cell-derived vesicles that contain various molecules including nucleic acids, proteins, and lipids. These vesicles can release their cargo into adjacent or distant cells and mediate intercellular communication and cellular function. Here we examined the regulation of epithelial sodium channels in mpkCCD cells and distal tubule Xenopus 2F3 cells by exosomes isolated from proximal tubule LLC-PK1 cells. Cultured mpkCCD cells were stained with CTX coupled to a green fluorophore in order to label the cell membranes and freshly isolated exosomes from LLC-PK1 cells were labeled with the red lipophilic dye PKH26 in order to visualize uptake of exosomes into the cells...
2016: PloS One
https://www.readbyqxmd.com/read/27796579/calcium-is-not-required-for-triggering-volume-restoration-in-hypotonically-challenged-a549-epithelial-cells
#12
Olga Ponomarchuk, Francis Boudreault, Sergei N Orlov, Ryszard Grygorczyk
Maintenance of cell volume is a fundamental housekeeping function in eukaryotic cells. Acute cell swelling activates a regulatory volume decrease (RVD) process with poorly defined volume sensing and intermediate signaling mechanisms. Here, we analyzed the putative role of Ca(2+) signaling in RVD in single substrate-adherent human lung epithelial A549 cells. Acute cell swelling was induced by perfusion of the flow-through imaging chamber with 50 % hypotonic solution at a defined fluid turnover rate. Changes in cytosolic Ca(2+) concentration ([Ca(2+)]i) and cell volume were monitored simultaneously with ratiometric Fura-2 fluorescence and 3D reconstruction of stereoscopic single-cell images, respectively...
November 2016: Pflügers Archiv: European Journal of Physiology
https://www.readbyqxmd.com/read/27743934/multiple-target-of-hamylin-on-rat-primary-hippocampal-neurons
#13
Nan Zhang, Shengchang Yang, Chang Wang, Jianghua Zhang, Lifang Huo, Yiru Cheng, Chuan Wang, Zhanfeng Jia, Leiming Ren, Lin Kang, Wei Zhang
Alzheimer's disease (AD) and type II diabetes mellitus (DM2) are the most common aging-related diseases and are characterized by β-amyloid and amylin accumulation, respectively. Multiple studies have indicated a strong correlation between these two diseases. Amylin oligomerization in the brain appears to be a novel risk factor for developing AD. Although amylin aggregation has been demonstrated to induce cytotoxicity in neurons through altering Ca(2+) homeostasis, the underlying mechanisms have not been fully explored...
February 2017: Neuropharmacology
https://www.readbyqxmd.com/read/27738490/functional-interactions-between-bkca%C3%AE-subunit-and-annexin-a5-implications-in-apoptosis
#14
Stephen P Brazier, Vsevolod Telezhkin, Paul J Kemp
Proteomic studies have suggested a biochemical interaction between α subunit of the large conductance, voltage- and Ca(2+)-activated potassium channel (BKCaα), and annexin A5 (ANXA5), which we verify here by coimmunoprecipitation and double labelling immunocytochemistry. The observation that annexin is flipped to the outer membrane leaflet of the plasma membrane during apoptosis, together with the knowledge that the intracellular C-terminal of BKCaα contains both Ca(2+)-binding and a putative annexin-binding motif, prompted us to investigate the functional consequences of this protein partnership to cell death...
2016: Oxidative Medicine and Cellular Longevity
https://www.readbyqxmd.com/read/27725751/cleaning-patch-clamp-pipettes-for-immediate-reuse
#15
I Kolb, W A Stoy, E B Rousseau, O A Moody, A Jenkins, C R Forest
Patch-clamp recording has enabled single-cell electrical, morphological and genetic studies at unparalleled resolution. Yet it remains a laborious and low-throughput technique, making it largely impractical for large-scale measurements such as cell type and connectivity characterization of neurons in the brain. Specifically, the technique is critically limited by the ubiquitous practice of manually replacing patch-clamp pipettes after each recording. To circumvent this limitation, we developed a simple, fast, and automated method for cleaning glass pipette electrodes that enables their reuse within one minute...
October 11, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27633787/accumulation-of-k-in-the-synaptic-cleft-modulates-activity-by-influencing-both-vestibular-hair-cell-and-calyx-afferent-in-the-turtle
#16
Donatella Contini, Steven D Price, Jonathan J Art
KEY POINTS: In the synaptic cleft between type I hair cells and calyceal afferents, K(+) ions accumulate as a function of activity, dynamically altering the driving force and permeation through ion channels facing the synaptic cleft. High-fidelity synaptic transmission is possible due to large conductances that minimize hair cell and afferent time constants in the presence of significant membrane capacitance. Elevated potassium maintains hair cells near a potential where transduction currents are sufficient to depolarize them to voltages necessary for calcium influx and synaptic vesicle fusion...
September 16, 2016: Journal of Physiology
https://www.readbyqxmd.com/read/27625130/-study-on-the-electrophsiological-properties-in-the-stria-vascularis-pericytes-in-cochlear-of-guinea-pig
#17
Y H Liu, Y P Wang, Y Wang, K T Ma, J Q Si, L Li
OBJECTIVE: The purpose of this paper was to study the electrophysiological properties and the type of potassium channels on cell membrane in the stria vascularis pericytes in cochlear of guinea pig. METHODS: Firstly examined the expression of the stria vascularis pericytes by desmin, a marker of pericytes, in cochlear of guinea pig with immunofluorescent method. Using whole-cell patch clamp recording techniques to observe electrophysiological properties in the cochlear pericytes in stria vascularis of guinea pig...
August 7, 2016: Zhonghua Er Bi Yan Hou Tou Jing Wai Ke za Zhi, Chinese Journal of Otorhinolaryngology Head and Neck Surgery
https://www.readbyqxmd.com/read/27605581/testosterone-decreases-urinary-bladder-smooth-muscle-excitability-via-novel-signaling-mechanism-involving-direct-activation-of-the-bk-channels
#18
Kiril L Hristov, Shankar P Parajuli, Aaron Provence, Georgi V Petkov
In addition to improving sexual function, testosterone has been reported to have beneficial effects in ameliorating lower urinary tract symptoms by increasing bladder capacity and compliance, while decreasing bladder pressure. However, the cellular mechanisms by which testosterone regulates detrusor smooth muscle (DSM) excitability have not been elucidated. Here, we used amphotericin-B perforated whole cell patch-clamp and single channel recordings on inside-out excised membrane patches to investigate the regulatory role of testosterone in guinea pig DSM excitability...
December 1, 2016: American Journal of Physiology. Renal Physiology
https://www.readbyqxmd.com/read/27572487/microdomain-specific-modulation-of-l-type-calcium-channels-leads-to-triggered-ventricular-arrhythmia-in-heart-failure
#19
Jose L Sanchez-Alonso, Anamika Bhargava, Thomas O'Hara, Alexey V Glukhov, Sophie Schobesberger, Navneet Bhogal, Markus B Sikkel, Catherine Mansfield, Yuri E Korchev, Alexander R Lyon, Prakash P Punjabi, Viacheslav O Nikolaev, Natalia A Trayanova, Julia Gorelik
RATIONALE: Disruption in subcellular targeting of Ca(2+) signaling complexes secondary to changes in cardiac myocyte structure may contribute to the pathophysiology of a variety of cardiac diseases, including heart failure (HF) and certain arrhythmias. OBJECTIVE: To explore microdomain-targeted remodeling of ventricular L-type Ca(2+) channels (LTCCs) in HF. METHODS AND RESULTS: Super-resolution scanning patch-clamp, confocal and fluorescence microscopy were used to explore the distribution of single LTCCs in different membrane microdomains of nonfailing and failing human and rat ventricular myocytes...
September 30, 2016: Circulation Research
https://www.readbyqxmd.com/read/27568501/hcn-channels-segregate-stimulation-evoked-movement-responses-in-neocortex-and-allow-for-coordinated-forelimb-movements-in-rodents
#20
Jeffery A Boychuk, Jordan S Farrell, Laura A Palmer, Anna C Singleton, Quentin J Pittman, G Campbell Teskey
KEY POINTS: The present study tested whether HCN channels contribute to the organization of motor cortex and to skilled motor behaviour during a forelimb reaching task. Experimental reductions in HCN channel signalling increase the representation of complex multiple forelimb movements in motor cortex as assessed by intracortical microstimulation. Global HCN1(KO) mice exhibit reduced reaching accuracy and atypical movements during a single-pellet reaching task relative to wild-type controls...
January 1, 2017: Journal of Physiology
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