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Single channel patch clamp

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https://www.readbyqxmd.com/read/29775559/mechanosensitivity-of-mitochondr-ial-large-conductance-calcium-activated-potassium-channels
#1
Agnieszka Walewska, Bogusz Kulawiak, Adam Szewczyk, Piotr Koprowski
Potassium channels have been discovered in the inner mitochondrial membrane of various cells. These channels can regulate the mitochondrial membrane potential, the matrix volume, respiration and reactive species generation. Therefore, it is believed that their activation is cytoprotective in various tissues. In our study, the single-channel activity of a large-conductance calcium-activated potassium channel (mitoBKCa ) was measured by the patch-clamp technique on mitoplasts derived from mitochondria isolated from human glioma U-87 MG cells...
May 15, 2018: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29768258/multiple-actions-of-rotenone-an-inhibitor-of-mitochondrial-respiratory-chain-on-ionic-currents-and-miniature-end-plate-potential-in-mouse-hippocampal-mhippoe-14-neurons
#2
Chin-Wei Huang, Kao-Min Lin, Te-Yu Hung, Yao-Chung Chuang, Sheng-Nan Wu
BACKGROUND/AIMS: Rotenone (Rot) is known to suppress the activity of complex I in the mitochondrial chain reaction; however, whether this compound has effects on ion currents in neurons remains largely unexplored. METHODS: With the aid of patch-clamp technology and simulation modeling, the effects of Rot on membrane ion currents present in mHippoE-14 cells were investigated. RESULTS: Addition of Rot produced an inhibitory action on the peak amplitude of INa with an IC50 value of 39...
May 11, 2018: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29758359/probing-flecainide-block-of-i-na-using-human-pluripotent-stem-cell-derived-ventricular-cardiomyocytes-adapted-to-automated-patch-clamping-and-2d-monolayers
#3
Lin Geng, Chi-Wing Kong, Andy O T Won, Angie Man-Yee Shum, Maggie Z Y Chow, Hui Che, Chenzi Zhang, Ka-Long Yau, Camie W Chan, Wendy Keung, Ronald A Li
Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are emerging tools for applications such as drug discovery and screening for pro-arrhythmogenicity and cardiotoxicity as leading causes for drug attrition. Understanding the electrophysiology (EP) of hPSC-CMs is essential but conventional manual patch-clamping is highly laborious and low-throughput. Here we adapted hPSC-CMs derived from two human embryonic stem cell (hESC) lines, HES2 and H7, for a 16-channel automated planar-recording approach for single-cell EP characterization...
May 11, 2018: Toxicology Letters
https://www.readbyqxmd.com/read/29752414/ligand-binding-and-activation-properties-of-the-purified-bacterial-cyclic-nucleotide-gated-channel-sthk
#4
Philipp A M Schmidpeter, Xiaolong Gao, Vikrant Uphadyay, Jan Rheinberger, Crina M Nimigean
Cyclic nucleotide-modulated ion channels play several essential physiological roles. They are involved in signal transduction in photoreceptors and olfactory sensory neurons as well as pacemaking activity in the heart and brain. Investigations of the molecular mechanism of their actions, including structural and electrophysiological characterization, are restricted by the availability of stable, purified protein obtained from accessible systems. Here, we establish that SthK, a cyclic nucleotide-gated (CNG) channel from Spirochaeta thermophila , is an excellent model for investigating the gating of eukaryotic CNG channels at the molecular level...
May 11, 2018: Journal of General Physiology
https://www.readbyqxmd.com/read/29733889/adenosine-promotes-endplate-nachr-channel-activity-in-adult-mouse-skeletal-muscle-fibres-via-low-affinity-p1-receptors
#5
Annalisa Bernareggi, Elisa Ren, Arthur Giniatullin, Elisa Luin, Marina Sciancalepore, Rashid Giniatullin, Paola Lorenzon
Adenosine is a powerful modulator of skeletal neuromuscular transmission, operating via inhibitory or facilitatory purinergic-type P1 receptors. To date, studies have been focused mainly on the effect of adenosine on presynaptic P1 receptors controlling transmitter release. In this study, using two-microelectrode voltage clamp and single channel patch-clamp recording techniques, we have explored potential postsynaptic targets of adenosine and their modulatory effect on nicotinic acetylcholine receptor (nAChR)-mediated synaptic responses in adult mouse skeletal muscle fibres in vitro...
May 4, 2018: Neuroscience
https://www.readbyqxmd.com/read/29716993/danger-associated-peptides-close-stomata-by-ost1-independent-activation-of-anion-channels-in-guard-cells
#6
Xiaojiang Zheng, Seock Kang, Yanping Jing, Zhijie Ren, Legong Li, Jian-Min Zhou, Gerald Berkowitz, Jisen Shi, Aigen Fu, Wenzhi Lan, Fugeng Zhao, Sheng Luan
The plant elicitor peptides (Peps), a family of damage/danger-associated molecular patterns (DAMPs), are perceived by two receptors, PEPR1 and PEPR2, and contribute to plant defense against pathogen attack and abiotic stress. Here we show that the Peps-PEPR signaling pathway functions in stomatal immunity by activating guard cell anion channels in Arabidopsis thaliana. The mutant plants lacking both PEPR1 and PEPR2 (pepr1 pepr2) displayed enhanced bacterial growth after being sprayed with Pseudomonas syringae pv...
April 30, 2018: Plant Cell
https://www.readbyqxmd.com/read/29697782/the-inhibitory-effect-of-bkca-channels-induced-by-autoantibodies-against-angiotensin-ii-type-1-receptor-is-independent-of-at1r
#7
Peng Wang, Suli Zhang, Jie Ren, Li Yan, Lina Bai, Li Wang, Pengli Wang, Jingwei Bian, Xiaochen Yin, Huirong Liu
Autoantibodies against angiotensin II Type 1 receptor (AT1-AA) are routinely detected in the serum of preeclampsia patients, which results in an increase in vascular tone and an elevation in intracellular calcium concentration of rat vascular smooth muscle (VSM). The big conductance calcium-activated potassium channels (BKCa channels) account for the dominant outward currents in VSMCs, contributing to membrane hyperpolarization and vasodilation. In the present study, we investigated the effect of AT1-AA on BKCa channels...
April 25, 2018: Acta Biochimica et Biophysica Sinica
https://www.readbyqxmd.com/read/29684406/conservation-of-cardiac-l-type-ca-2-channels-and-their-regulation-in-drosophila-a-novel-genetically-pliable-channelopathic-model
#8
Worawan B Limpitikul, Meera C Viswanathan, Brian O'Rourke, David T Yue, Anthony Cammarato
Dysregulation of L-type Ca2+ channels (LTCCs) underlies numerous cardiac pathologies. Understanding their modulation with high fidelity relies on investigating LTCCs in their native environment with intact interacting proteins. Such studies benefit from genetic manipulation of endogenous channels in cardiomyocytes, which often proves cumbersome in mammalian models. Drosophila melanogaster, however, offers a potentially efficient alternative as it possesses a relatively simple heart, is genetically pliable, and expresses well-conserved genes...
April 20, 2018: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/29674316/activation-of-renal-clc-k-chloride-channels-depends-on-an-intact-n-terminus-of-their-accessory-subunit-barttin
#9
Daniel Wojciechowski, Stefan Thiemann, Christina Schaal, Alina Rahtz, Jeanne de la Roche, Birgit Begemann, Toni Becher, Martin Fischer
ClC-K channels belong to the CLC family of chloride channels and chloride/proton antiporters. They contribute to sodium chloride reabsorption in Henle's loop of the kidney and to potassium secretion into the endolymph by the stria vascularis of the inner ear. Their accessory subunit barttin stabilizes the ClC-K/barttin complex, promotes its insertion into the surface membrane, and turns the pore-forming subunits into a conductive state. Barttin mutations cause Bartter syndrome type IV, a salt wasting nephropathy with sensorineural deafness...
April 19, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29671361/liposomal-quercetin-potentiates-maxi-k-channel-openings-in-smooth-muscles-and-restores-its-activity-after-oxidative-stress
#10
Mariia I Melnyk, Dariia O Dryn, Lina T Al Kury, Alexander V Zholos, Anatoly I Soloviev
The effects of quercetin-loaded liposomes (PCL-Q) and their constituents, that is, free quercetin (Q) and 'empty' phosphatidylcholine vesicles (PCL), on maxi-K channel activity were studied in single mouse ileal myocytes before and after H2 O2 -induced oxidative stress. Macroscopic Maxi-K channel currents were recorded using whole-cell patch clamp techniques, while single BKCa channel currents were recorded in the cell-attached configuration. Bath application of PCL-Q (100 μg/ml of lipid and 3 μg/ml of quercetin) increased single Maxi-K channel activity more than threefold, from 0...
April 19, 2018: Journal of Liposome Research
https://www.readbyqxmd.com/read/29650545/regulation-of-blood-pressure-by-targeting-ca-v-1-2-galectin-1-protein-interaction
#11
Zhenyu Hu, Guang Li, Jiong-Wei Wang, Suet Yen Chong, Dejie Yu, Xiaoyuan Wang, Jia Lin Soon, Mui Cheng Liang, Yuk Peng Wong, Na Huang, Henry M Colecraft, Ping Liao, Tuck Wah Soong
Background -L-type CaV 1.2 channels play crucial roles in regulation of blood pressure. Galectin-1 (Gal-1), has been reported to bind to the I-II loop of CaV 1.2 channels to reduce their current density. However, the mechanistic understanding for the down-regulation of CaV 1.2 channels by Gal-1, and whether Gal-1 plays a direct role in blood pressure regulation remain unclear. Methods - In vitro experiments involving co-IP, western blot, patch-clamp recordings, immunohistochemistry and pressure myography were used to evaluate the molecular mechanisms by which Gal-1 down-regulates CaV 1...
April 12, 2018: Circulation
https://www.readbyqxmd.com/read/29589117/the-degenerin-region-of-the-human-bile-acid-sensitive-ion-channel-basic-is-involved-in-channel-inhibition-by-calcium-and-activation-by-bile-acids
#12
Alexandr V Ilyaskin, Sonja A Kirsch, Rainer A Böckmann, Heinrich Sticht, Christoph Korbmacher, Silke Haerteis, Alexei Diakov
The bile acid-sensitive ion channel (BASIC) is a member of the ENaC/degenerin family of ion channels. It is activated by bile acids and inhibited by extracellular Ca2+ . The aim of this study was to explore the molecular mechanisms mediating these effects. The modulation of BASIC function by extracellular Ca2+ and tauro-deoxycholic acid (t-DCA) was studied in Xenopus laevis oocytes heterologously expressing human BASIC using the two-electrode voltage-clamp and outside-out patch-clamp techniques. Substitution of aspartate D444 to alanine or cysteine in the degenerin region of BASIC, a region known to be critically involved in channel gating, resulted in a substantial reduction of BASIC Ca2+ sensitivity...
March 27, 2018: Pflügers Archiv: European Journal of Physiology
https://www.readbyqxmd.com/read/29588365/a-three-way-inter-molecular-network-accounts-for-the-ca-v-%C3%AE-2%C3%AE-1-induced-functional-modulation-of-the-pore-forming-ca-v-1-2-subunit
#13
Julie Briot, Olivier Mailhot, Benoîte Bourdin, Marie-Philippe Tétreault, Rafael Najmanovich, Lucie Parent
L-type CaV 1.2 channels are essential for the excitation-contraction coupling in cardiomyocytes and are hetero-oligomers of a pore-forming CaV α1C assembled with CaV β and CaV α2δ1 subunits. A direct interaction between CaV α2δ1 and Asp-181 in the first extracellular loop of CaV α1 reproduces the native properties of the channel. A 3-D model of the von Willebrand factor type A (VWA) domain of CaV α2δ1 complexed with the voltage sensor domain of CaV α1C suggests that Ser-261 and Ser-263 residues in the metal ion-dependent adhesion site (MIDAS) motif are determinant in this interaction, but this hypothesis is untested...
March 27, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29554219/prrt2-controls-neuronal-excitability-by-negatively-modulating-na-channel-1-2-1-6-activity
#14
Floriana Fruscione, Pierluigi Valente, Bruno Sterlini, Alessandra Romei, Simona Baldassari, Manuela Fadda, Cosimo Prestigio, Giorgia Giansante, Jacopo Sartorelli, Pia Rossi, Alicia Rubio, Antonio Gambardella, Thierry Nieus, Vania Broccoli, Anna Fassio, Pietro Baldelli, Anna Corradi, Federico Zara, Fabio Benfenati
See Lerche (doi:10.1093/brain/awy073) for a scientific commentary on this article.Proline-rich transmembrane protein 2 (PRRT2) is the causative gene for a heterogeneous group of familial paroxysmal neurological disorders that include seizures with onset in the first year of life (benign familial infantile seizures), paroxysmal kinesigenic dyskinesia or a combination of both. Most of the PRRT2 mutations are loss-of-function leading to haploinsufficiency and 80% of the patients carry the same frameshift mutation (c...
April 1, 2018: Brain: a Journal of Neurology
https://www.readbyqxmd.com/read/29545386/%C3%AE-2-adrenergic-receptor-activation-promotes-long-term-potentiation-at-excitatory-synapses-in-the-mouse-accessory-olfactory-bulb
#15
Guang-Zhe Huang, Mutsuo Taniguchi, Ye-Bo Zhou, Jing-Ji Zhang, Fumino Okutani, Yoshihiro Murata, Masahiro Yamaguchi, Hideto Kaba
The formation of mate recognition memory in mice is associated with neural changes at the reciprocal dendrodendritic synapses between glutamatergic mitral cell (MC) projection neurons and GABAergic granule cell (GC) interneurons in the accessory olfactory bulb (AOB). Although noradrenaline (NA) plays a critical role in the formation of the memory, the mechanism by which it exerts this effect remains unclear. Here we used extracellular field potential and whole-cell patch-clamp recordings to assess the actions of bath-applied NA (10 µM) on the glutamatergic transmission and its plasticity at the MC-to-GC synapse in the AOB...
April 2018: Learning & Memory
https://www.readbyqxmd.com/read/29530619/o-glcnacylation-of-cardiac-nav1-5-contributes-to-the-development-of-arrhythmias-in-diabetic-hearts
#16
Peng Yu, Lili Hu, Jinyan Xie, Sisi Chen, Lin Huang, Zixuan Xu, Xiao Liu, Qiongqiong Zhou, Ping Yuan, Xia Yan, Jiejin Jin, Yang Shen, Wengen Zhu, Linghua Fu, Qi Chen, Jianhua Yu, Jianxin Hu, Qing Cao, Rong Wan, Kui Hong
BACKGROUND: Cardiovascular complications are major causes of mortality and morbidity in diabetic patients. The mechanisms underlying the progression of diabetic heart (DH) to ventricular arrhythmias are unclear. O-linked GlcNAcylation (O-GlcNAc) is a reversible post-translational modification for the regulation of diverse cellular processes. The purpose of this study was to assess whether the cardiac voltage-gated sodium channel (Nav1.5) is subjected to O-linked GlcNAcylation (O-GlcNAc), which plays an essential role in DH-induced arrhythmias...
June 1, 2018: International Journal of Cardiology
https://www.readbyqxmd.com/read/29524421/a-mechanically-activated-trpc1-like-current-in-white-adipocytes
#17
Mickaël F El Hachmane, Charlotta S Olofsson
Ca2+ impacts a large array of cellular processes in every known cell type. In the white adipocyte, Ca2+ is involved in regulation of metabolic processes such as lipolysis, glucose uptake and hormone secretion. Although the importance of Ca2+ in control of white adipocyte function is clear, knowledge is still lacking regarding the control of dynamic Ca2+ alterations within adipocytes and mechanisms inducing intracellular Ca2+ changes remain elusive. Own work has recently demonstrated the existence of store-operated Ca2+ entry (SOCE) in lipid filled adipocytes...
April 15, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29517089/revealing-dynamically-organized-receptor-ion-channel-clusters-in-live-cells-by-a-correlated-electric-recording-and-super-resolution-single-molecule-imaging-approach
#18
Rajeev Yadav, H Peter Lu
The N-methyl-d-aspartate (NMDA) receptor ion-channel is activated by the binding of ligands, along with the application of action potential, important for synaptic transmission and memory functions. Despite substantial knowledge of the structure and function, the gating mechanism of the NMDA receptor ion channel for electric on-off signals is still a topic of debate. We investigate the NMDA receptor partition distribution and the associated channel's open-close electric signal trajectories using a combined approach of correlating single-molecule fluorescence photo-bleaching, single-molecule super-resolution imaging, and single-channel electric patch-clamp recording...
March 28, 2018: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/29494673/the-bovine-trpv3-as-a-pathway-for-the-uptake-of-na-ca2-and-nh4
#19
Katharina T Schrapers, Gerhard Sponder, Franziska Liebe, Hendrik Liebe, Friederike Stumpff
Absorption of ammonia from the gastrointestinal tract results in problems that range from hepatic encephalopathy in humans to poor nitrogen efficiency of cattle with consequences for the global climate. Previous studies on epithelia and cells from the native ruminal epithelium suggest functional involvement of the bovine homologue of TRPV3 (bTRPV3) in ruminal NH4+ transport. Since the conductance of TRP channels to NH4+ has never been studied, bTRPV3 was overexpressed in HEK-293 cells and investigated using the patch-clamp technique and intracellular calcium imaging...
2018: PloS One
https://www.readbyqxmd.com/read/29493159/-effects-of-isovitexin-on-transient-outward-potassium-current-of-ventricular-myocytes-in-rats
#20
Jing-Na Ren, Ben-Long Rao, Hong-Xin Ma, Mao-Mao Sha, Yi Kuang, Zheng-Xin Xu
To investigate the effects of isovitexin Ⅳ on transient outward potassium current in rat ventricular myocytes. In this study, MTT assay was used to investigate the safe range of isovitexin. The results showed that the IC₅₀ of the drug was in the range of 10-30 μmol•L⁻¹, and the drug concentration of 1-3 μmol•L⁻¹ for the patch clamp test was within the safe range. In addition, the single ventricular myocytes were obtained by single-enzymatic hydrolysis through aortic retrograde perfusion. The transient outward potassium current (Ito) of rat ventricular myocytes was guided and measured by whole-cell patch-clamp technique and the changes of current characteristics were recorded after isovite was applied...
December 2017: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
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