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Single channel patch clamp

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https://www.readbyqxmd.com/read/29150838/gi-530159-a-novel-selective-mechano-sensitive-k2p-channel-opener-reduces-rat-dorsal-root-ganglion-drg-neuron-excitability
#1
Alexandre J C Loucif, Pierre-Philippe Saintot, Jia Liu, Brett M Antonio, Shannon G Zellmer, Katrina Yoger, Emma L Veale, Anna Wilbrey, Kiyoyuki Omoto, Lishuang Cao, Alex Gutteridge, Neil A Castle, Edward B Stevens, Alistair Mathie
BACKGROUND AND PURPOSE: TREK two pore domain potassium channels play a critical role in regulating the excitability of somatosensory nociceptive neurons and are important mediators of pain perception. An understanding of the roles of TREK channels in pain perception and, indeed, in other pathophysiological conditions, has been severely hampered by the lack of potent and/or selective activators and inhibitors. In this study we describe a new, selective opener of TREK channels, GI-530159 (4,4'-(Hexafluoroisopropylidene)bis(p-phenyleneoxy)dianiline)...
November 18, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/29143966/drug-interaction-at-herg-channel-in-vitro-assessment-of-the-electrophysiological-consequences-of-drug-combinations-and-comparison-against-theoretical-models
#2
Barbara Wiśniowska, Bartosz Lisowski, Magdalena Kulig, Sebastian Polak
Drugs carry a proarrhythmic risk, which gets even greater when they are used in combination. In vitro assessment of the proarrhythmic potential of drugs is limited to one compound and thus neglects the potential of drug-drug interactions, including those involving active metabolites. Here we present the results of an in vitro study of potential drug-drug interactions at the level of the hERG channel for the combination of up to three compounds: loratadine, desloratadine and ketoconazole. Experiments were performed at room temperature on an automated patch-clamp device CytoPatch 2, with the use of heterogeneously, stably transfected HEK cells...
November 16, 2017: Journal of Applied Toxicology: JAT
https://www.readbyqxmd.com/read/29134279/dietary-k-and-cl-independently-regulate-basolateral-conductance-in-principal-and-intercalated-cells-of-the-collecting-duct
#3
Viktor N Tomilin, Oleg Zaika, Arohan R Subramanya, Oleh Pochynyuk
The renal collecting duct contains two distinct cell types, principal and intercalated cells, expressing potassium Kir4.1/5.1 (KCNJ10/16) and chloride ClC-K2 (ClC-Kb in humans) channels on their basolateral membrane, respectively. Both channels are thought to play important roles in controlling systemic water-electrolyte balance and blood pressure. However, little is known about mechanisms regulating activity of Kir4.1/5.1 and ClC-K2/b. Here, we employed patch clamp analysis at the single channel and whole cell levels in freshly isolated mouse collecting ducts to investigate regulation of Kir4...
November 13, 2017: Pflügers Archiv: European Journal of Physiology
https://www.readbyqxmd.com/read/29118035/single-channel-analysis-of-isoflurane-and-ethanol-enhancement-of-taurine-activated-glycine-receptors
#4
Dean Kirson, Jelena Todorovic, S John Mihic
The amino acid taurine is an endogenous ligand acting on glycine receptors and is released by astrocytes in many brain regions, such as the nucleus accumbens and prefrontal cortex. Taurine is a partial agonist with an efficacy significantly lower than that of glycine. Allosteric modulators such as ethanol and isoflurane produce leftward shifts of glycine concentration-response curves but have no effects at saturating glycine concentrations. In contrast, in whole-cell electrophysiology studies these modulators increase the effects of saturating taurine concentrations...
November 8, 2017: Journal of Pharmacology and Experimental Therapeutics
https://www.readbyqxmd.com/read/29107716/pacsin-2-is-required-for-the-maintenance-of-a-normal-cardiac-function-in-the-developing-mouse-heart
#5
Judith Semmler, Jan Kormann, Sureshkumar Perumal Srinivasan, Annette Köster, Daniel Sälzer, Michael Reppel, Jürgen Hescheler, Markus Plomann, Filomain Nguemo
The Pacsin proteins (Pacsin 1, 2 and 3) play an important role in intracellular trafficking and thereby signal transduction in many cells types. This study was designed to examine the role of Pacsin 2 in cardiac development and function. We investigated the development and electrophysiological properties of Pacsin 2 knockout (P2KO) hearts and single cardiomyocytes isolated from 11.5 and 15.5days old fetal mice. Immunofluorescence experiments confirmed the lack of Pacsin 2 protein expression in P2KO cardiac myocytes in comparison to wildtype (WT)...
October 28, 2017: Pharmacological Research: the Official Journal of the Italian Pharmacological Society
https://www.readbyqxmd.com/read/29090695/on-the-selectivity-of-the-nachbac-channel-an-integrated-computational-and-experimental-analysis-of-sodium-and-calcium-permeation
#6
Carlo Guardiani, Olena A Fedorenko, Stephen K Roberts, Igor A Khovanov
Ion channel selectivity is essential for their function, yet the molecular basis of a channel's ability to select between ions is still rather controversial. In this work, using a combination of molecular dynamics simulations and electrophysiological current measurements we analyze the ability of the NaChBac channel to discriminate between calcium and sodium. Our simulations show that a single calcium ion can access the Selectivity Filter (SF) interacting so strongly with the glutamate ring so as to remain blocked inside...
November 1, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/29089441/glutamate-activates-ampa-receptor-conductance-in-the-developing-schwann-cells-of-the-mammalian-peripheral-nerves
#7
Ting-Jiun Chen, Nicole Fröhlich, Bartosz Kula, Ruxandra Barzan, Maria Kukley
Schwann cells (SCs) are myelinating cells of the peripheral nervous system. Although SCs are known to express different channels and receptors on their surface, little is known about the activation and function of these proteins. Ionotropic glutamate receptors are thought to play an essential role during development of SC lineage and during peripheral nerve injury and therefore we sought to study their functional properties. We established a novel preparation of living peripheral nerve slices with preserved cellular architecture, and used a patch-clamp technique to study α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic-acid (AMPA)-receptor-mediated currents in SCs for the first time...
October 31, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/29074990/organelle-membrane-derived-patches-reshaping-classical-methods-for-new-targets
#8
George Shapovalov, Abigaël Ritaine, Gabriel Bidaux, Christian Slomianny, Anne-Sophie Borowiec, Dmitri Gordienko, Geert Bultynck, Roman Skryma, Natalia Prevarskaya
Intracellular ion channels are involved in multiple signaling processes, including such crucial ones as regulation of cellular motility and fate. With 95% of the cellular membrane belonging to intracellular organelles, it is hard to overestimate the importance of intracellular ion channels. Multiple studies have been performed on these channels over the years, however, a unified approach allowing not only to characterize their activity but also to study their regulation by partner proteins, analogous to the patch clamp "golden standard", is lacking...
October 26, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29054409/dual-expression-of-constitutively-active-g%C3%AE-s-protein-coupled-receptors-differentially-establishes-the-resting-activity-of-the-camp-gated-hcn2-channel-in-a-single-compartment
#9
Noriyuki Nakashima, Kie Nakashima, Takeo Nakayama, Akiko Takaku, Ryosuke Kanamori
The hyperpolarization-activated cyclic nucleotide-gated 2 (HCN2) channel is a major subtype of the HCN channel family expressed in the nervous system that sets the membrane potential, regulates cell excitability and senses changes in the extracellular environment. Neurons express various Gαs-protein-coupled receptors (GPCRs), many of which show ligand-independent constitutive activity. These membrane-bound proteins are expressed in various subcellular compartments of neurons. Therefore, some proportion of HCN2 channels opens in response to the basal cAMP pool size produced by constitutively active GPCRs...
December 9, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29046423/the-intrinsic-physiology-of-inhibitory-neurons-changes-over-auditory-development
#10
Briana J Carroll, Richard Bertram, Richard L Hyson
During auditory development, changes in membrane properties promote the ability of excitatory neurons in the brainstem to code aspects of sound, including the level and timing of a stimulus. Some of these changes coincide with hearing onset, suggesting that sound-driven neural activity produces developmental plasticity of ion channel expression. While it is known that the coding properties of excitatory neurons are modulated by inhibition in the mature system, it is unknown whether there are also developmental changes in the membrane properties of brainstem inhibitory neurons...
October 18, 2017: Journal of Neurophysiology
https://www.readbyqxmd.com/read/29016681/heterodimerization-of-two-pore-domain-k-channel-task1-and-talk2-in-living-heterologous-expression-systems
#11
Yoshiaki Suzuki, Kanako Tsutsumi, Tatsuya Miyamoto, Hisao Yamamura, Yuji Imaizumi
Two-pore-domain K+ (K2P) channels sense a wide variety of stimuli such as mechanical stress, inhalational anesthetics, and changes in extracellular pH or temperature. The K2P channel activity forms a background K+ current and, thereby, contributes to resting membrane potentials. Six subfamilies including fifteen subtypes of K2P channels have been identified. Each K2P channel molecule with two pores consists of a homodimer of each subtype. In addition, a few heterodimers mainly within the same subfamilies have been found recently...
2017: PloS One
https://www.readbyqxmd.com/read/28993909/coexistence-of-clcn1-and-scn4a-mutations-in-one-family-suffering-from-myotonia
#12
Lorenzo Maggi, Sabrina Ravaglia, Alessandro Farinato, Raffaella Brugnoni, Concetta Altamura, Paola Imbrici, Diana Conte Camerino, Alessandro Padovani, Renato Mantegazza, Pia Bernasconi, Jean-François Desaphy, Massimiliano Filosto
Non-dystrophic myotonias are characterized by clinical overlap making it challenging to establish genotype-phenotype correlations. We report clinical and electrophysiological findings in a girl and her father concomitantly harbouring single heterozygous mutations in SCN4A and CLCN1 genes. Functional characterization of N1297S hNav1.4 mutant was performed by patch clamp. The patients displayed a mild phenotype, mostly resembling a sodium channel myotonia. The CLCN1 c.501C>G (p.F167L) mutation has been already described in recessive pedigrees, whereas the SCN4A c...
October 9, 2017: Neurogenetics
https://www.readbyqxmd.com/read/28982976/activation-of-the-phospholipid-scramblase-tmem16f-by-nanosecond-pulsed-electric-field-nspef-facilitates-its-diverse-cytophysiological-effects
#13
Claudia Muratori, Andrei G Pakhomov, Elena Gianulis, Jade Meads, Maura Casciola, Peter A Mollica, Olga N Pakhomova
Nanosecond pulsed electric fields (nsPEFs) are emerging as a novel modality for cell stimulation and tissue ablation. However, the downstream protein effectors responsible for nsPEF bioeffects remain to be established. Here we demonstrate that nsPEFs activate TMEM16F, (or anoctamin 6), a protein functioning as a Ca(2+)-dependent phospholipid scramblase and Ca(2+)-activated chloride channel. Using confocal microscopy and patch-clamp recording, we investigated the relevance of TMEM16F activation for several bioeffects triggered by nsPEF, including phosphatidylserine (PS) externalization, nanopore-conducted currents, membrane blebbing, and cell death...
October 5, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28974519/delayed-repolarization-underlies-ventricular-arrhythmias-in-rats-with-heart-failure-and-preserved-ejection-fraction
#14
Jae Hyung Cho, Rui Zhang, Peter J Kilfoil, Romain Gallet, Geoffrey de Couto, Catherine Bresee, Joshua I Goldhaber, Eduardo Marbán, Eugenio Cingolani
Background -Heart failure with preserved ejection fraction (HFpEF) represents approximately half of heart failure, and its incidence continues to increase. The leading cause of mortality in HFpEF is sudden death, but little is known about the underlying mechanisms. Methods -Dahl salt-sensitive rats were fed a high-salt diet (8% NaCl) from 7 weeks of age to induce HFpEF (n=38). Rats fed a normal-salt diet (0.3% NaCl) served as controls (n=13). Echocardiograms were performed to assess systolic and diastolic function from 14 weeks of age...
October 3, 2017: Circulation
https://www.readbyqxmd.com/read/28938737/complex-catsper-dependent-and-independent-ca2-i-signalling-in-human-spermatozoa-induced-by-follicular-fluid
#15
Sean G Brown, Sarah Costello, Mark C Kelly, Mythili Ramalingam, Ellen Drew, Stephen J Publicover, Christopher L R Barratt, Sarah Martins Da Silva
STUDY QUESTION: Does progesterone in human follicular fluid (hFF) activate CatSper and do other components of hFF modulate this effect and/or contribute separately to hFF-induced Ca2+ signaling? SUMMARY ANSWER: hFF potently stimulates CatSper and increases [Ca2+]i, primarily due to high concentrations of progesterone, however, other components of hFF also contribute to [Ca2+]i signaling, including modulation of CatSper channel activity and inhibition of [Ca2+]i oscillations...
October 1, 2017: Human Reproduction
https://www.readbyqxmd.com/read/28936725/ampa-receptors-at-ribbon-synapses-in-the-mammalian-retina-kinetic-models-and-molecular-identity
#16
Espen Hartveit, Bas-Jan Zandt, Eirik Madsen, Áurea Castilho, Svein Harald Mørkve, Margaret Lin Veruki
In chemical synapses, neurotransmitter molecules released from presynaptic vesicles activate populations of postsynaptic receptors that vary in functional properties depending on their subunit composition. Differential expression and localization of specific receptor subunits are thought to play fundamental roles in signal processing, but our understanding of how that expression is adapted to the signal processing in individual synapses and microcircuits is limited. At ribbon synapses, glutamate release is independent of action potentials and characterized by a high and rapidly changing rate of release...
September 21, 2017: Brain Structure & Function
https://www.readbyqxmd.com/read/28900735/4-chloro-3-nitro-n-butylbenzenesulfonamide-acts-on-kv3-1-channels-by-an-open-channel-blocker-mechanism
#17
Carlos Alberto Zanutto Bassetto Junior, Wamberto Antonio Varanda, Eduardo René Pérez González
The effects of 4-chloro-3-nitro-N-butylbenzenesulfonamide (SMD2) on KV3.1 channels, heterologous expressed in L-929 cells, were studied with the whole cell patch-clamp technique. SMD2 blocks KV3.1 in a reversible and use-dependent manner, with IC50 around 10 µM, and a Hill coefficient around 2. Although the conductance vs. voltage relationship in control condition can be described by a single Boltzmann function, two terms are necessary to describe the data in the presence of SMD2. The activation and deactivation time constants are weakly voltage dependent both for control and in the presence of SMD2...
November 2017: Amino Acids
https://www.readbyqxmd.com/read/28893911/molecular-and-functional-characterization-of-the-endothelial-atp-sensitive-potassium-channel
#18
Qadeer Aziz, Yiwen Li, Naomi Anderson, Leona Ojake, Elena Tsisanova, Andrew Tinker
ATP-sensitive potassium (KATP) channels are widely expressed in the cardiovascular system, where they regulate a range of biological activities by linking cellular metabolism with membrane excitability. KATP channels in vascular smooth muscle have a well-defined role in regulating vascular tone. KATP channels are also thought to be expressed in vascular endothelial cells, but their presence and function in this context are less clear. As a result, we aimed to investigate the molecular composition and physiological role of endothelial KATP channels...
October 27, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28885682/selective-targeting-of-m-type-potassium-kv-7-4-channels-demonstrates-their-key-role-in-the-regulation-of-dopaminergic-neuronal-excitability-and-depression-like-behaviour
#19
Li Li, Hui Sun, Jie Ding, Chenxu Niu, Min Su, Ludi Zhang, Yingmin Li, Chuan Wang, Nikita Gamper, Xiaona Du, Hailin Zhang
BACKGROUND AND PURPOSE: The mesolimbic dopamine system originating in the ventral tegmental area (VTA) is involved in the development of depression, and firing patterns of VTA dopaminergic neurons are key determinants in this process. Here, we describe a crucial role for the M-type Kv 7.4 channels in modulating excitability of VTA dopaminergic neurons and in the development of depressive behaviour in mice. EXPERIMENTAL APPROACH: We used Kv 7.4 channel knockout mice and a social defeat model of depression in combination with electrophysiological techniques (patch clamp recording and in vivo single-unit recordings), immunohistochemistry, single-cell PCR and behavioural analyses (social interaction time and glucose preference tests) to investigate VTA circuits involved in the development of depression-like behaviour...
September 8, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28882752/mechanosensitive-ion-channels-in-articular-nociceptors-drive%C3%A2-mechanical-allodynia-in-osteoarthritis
#20
B H He, M Christin, S Mouchbahani-Constance, A Davidova, R Sharif-Naeini
OBJECTIVE: Osteoarthritis (OA) is a disabling and highly prevalent condition affecting millions worldwide. Pain is the major complaint of OA patients and is presently inadequately managed. It manifests as mechanical allodynia, a painful response to innocuous stimuli such as joint movement. Allodynia is due in part to the sensitization of articular nociceptors to mechanical stimuli. These nociceptors respond to noxious mechanical stimuli applied to their terminals via the expression of depolarizing high-threshold mechanosensitive ion channels (MSICs) that convert painful mechanical forces into electrical signals...
September 5, 2017: Osteoarthritis and Cartilage
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