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Whole cell patch clamp

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https://www.readbyqxmd.com/read/28419410/selective-and-state-dependent-activation-of-tresk-background-potassium-channel-by-cloxyquin
#1
Miklós Lengyel, Alice Dobolyi, Gábor Czirják, Péter Enyedi
BACKGROUND AND PURPOSE: Cloxyquin (5-cloroquinolin-8-ol) has been previously described as an activator of TRESK (K2P 18.1, TWIK-related spinal cord K(+) channel) background potassium channel. We have examined the specificity of the drug by testing several K2P channels. We have investigated the mechanism of cloxyquin-mediated TRESK activation, focusing on the differences between the physiologically relevant regulatory states of the channel. EXPERIMENTAL APPROACH: Potassium currents were measured by two-electrode voltage clamp in Xenopus oocytes and by whole-cell patch clamp in mouse dorsal root ganglion (DRG) neurons...
April 18, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28417339/expression-and-function-of-the-small-conductance-ca-2-activated-k-channel-is-decreased-in-urinary-bladder-smooth-muscle-cells-from-female-guinea-pig-with-partial-bladder-outlet-obstruction
#2
Ning Li, Honglin Ding, Xiaoning He, Zizheng Li, Yili Liu
PURPOSE: Overactive bladder (OAB), usually accompanied by partial bladder outlet obstruction (PBOO), is associated with detrusor overactivity (DO) which is related to the increased urinary bladder smooth muscle (UBSM) cells excitability. Small-conductance Ca(2+)-activated K(+) (SK) channels play a constitutive regulatory role of UBSM excitability and contractility. PBOO is associated with the decreased SK channels mRNA expression and the attenuated regulative effect of SK channels on UBSM contractility...
April 17, 2017: International Urology and Nephrology
https://www.readbyqxmd.com/read/28416807/a-simplified-protocol-for-differentiation-of-electrophysiologically-mature-neuronal-networks-from-human-induced-pluripotent-stem-cells
#3
N Gunhanlar, G Shpak, M van der Kroeg, L A Gouty-Colomer, S T Munshi, B Lendemeijer, M Ghazvini, C Dupont, W J G Hoogendijk, J Gribnau, F M S de Vrij, S A Kushner
Progress in elucidating the molecular and cellular pathophysiology of neuropsychiatric disorders has been hindered by the limited availability of living human brain tissue. The emergence of induced pluripotent stem cells (iPSCs) has offered a unique alternative strategy using patient-derived functional neuronal networks. However, methods for reliably generating iPSC-derived neurons with mature electrophysiological characteristics have been difficult to develop. Here, we report a simplified differentiation protocol that yields electrophysiologically mature iPSC-derived cortical lineage neuronal networks without the need for astrocyte co-culture or specialized media...
April 18, 2017: Molecular Psychiatry
https://www.readbyqxmd.com/read/28415676/nitrobenzylthioinosine-mimics-adenosine-to-attenuate-the-epileptiform-discharge-of-hippocampal-neurons-from-epileptic-rats
#4
Hao Huang, Jing Wang, Jun Zhang, Zhong Luo, Dongxu Li, Xiaowei Qiu, Yan Peng, Zhongxiang Xu, Ping Xu, Zucai Xu
Nitrobenzylthioinosine (NBTI), a specific inhibitor of type 1 equilibrative nucleoside transporter, could regulate the extracellular adenosine concentration and have protective roles in seizures. However, the protection mechanism of NBTI in seizures remains poorly understood. Here, the expression pattern and subcellular distribution of adenosine A1 receptor were detected by Western blot analysis and double-labeling immunofluorescence staining in Lithium Chloride-Pilocarpine induced epileptic rat model. At 24 h after pilocarpine induced rat seizures, hippocampal slices were prepared and the evoked excitatory postsynaptic currents (eEPSCs) amplitude of pyramidal neurons in hippocampus CA1 region was recorded using whole-cell patch clamp...
March 8, 2017: Oncotarget
https://www.readbyqxmd.com/read/28411554/emodin-suppresses-the-nasopharyngeal-carcinoma-cells-by-targeting-the-chloride-channels
#5
Lianshun Ma, Yaping Yang, Zizhang Yin, Mei Liu, Liwei Wang, Lixin Chen, Linyan Zhu, Haifeng Yang
Emodin is a natural anthraquinone derivative isolated from the Rheum palmatum. Recent studies demonstrated that emodin has anti-cancer activity in different kinds of human cancer cell lines. However, the underlying mechanism has not been very well studied. Our previous studies showed chloride channels is an important target of anti-cancer drugs. Therefore, the purpose of this research was aimed to explore the role of chloride channels involving in the anti-cancer activity of emodin. The proliferation, cell cycle arrest and apoptosis of poorly differentiated human nasopharyngeal carcinoma cells (CNE-2Z) and normal nasopharyngeal epithelial cells (NP69-SV40T) were detected by 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide(MTT)and flow cytometry...
April 12, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28411183/membrane-permeation-of-arginine-rich-cell-penetrating-peptides-independent-of-transmembrane-potential-as-a-function-of-lipid-composition-and-membrane-fluidity
#6
Rike Wallbrecher, Tobias Ackels, R Alis Olea, Marco J Klein, Lucie Caillon, Jürgen Schiller, Petra H Bovée-Geurts, Toin H van Kuppevelt, Anne S Ulrich, Marc Spehr, Merel J W Adjobo-Hermans, Roland Brock
Cell-penetrating peptides (CPPs) are prominent delivery vehicles to confer cellular entry of (bio-) macromolecules. Internalization efficiency and uptake mechanism depend, next to the type of CPP and cargo, also on cell type. Direct penetration of the plasma membrane is the preferred route of entry as this circumvents endolysosomal sequestration. However, the molecular parameters underlying this import mechanism are still poorly defined. Here, we make use of the frequently used HeLa and HEK cell lines to address the role of lipid composition and membrane potential...
April 11, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28411158/intrinsic-membrane-properties-and-cholinergic-modulation-of-mouse-basal-forebrain-glutamatergic-neurons-in-vitro
#7
Chun Yang, James T McKenna, Ritchie E Brown
The basal forebrain (BF) controls sleep-wake cycles, attention and reward processing. Compared to cholinergic and GABAergic neurons, BF glutamatergic neurons are less well understood, due to difficulties in identification. Here, we use vesicular glutamate transporter 2 (vGluT2)-tdTomato mice, expressing a red fluorescent protein (tdTomato) in the major group of BF glutamatergic neurons (vGluT2+) to characterize their intrinsic electrical properties and cholinergic modulation. Whole-cell, patch-clamp recordings were made from vGluT2+ neurons in coronal BF slices...
April 12, 2017: Neuroscience
https://www.readbyqxmd.com/read/28399683/kcnq2-associated-neonatal-epilepsy-phenotype-might-correlate-with-genotype
#8
Inn-Chi Lee, Jiann-Jou Yang, Jao-Shwann Liang, Tung-Ming Chang, Shuan-Yow Li
We analyzed the KCNQ2 wild-type gene and 3 mutations to highlight the important association between the KCNQ2 phenotype and genotype. The clinical phenotypes of 3 mutations (p.E515D, p.V543 M, and p.R213Q) were compared. KCNQ2, wild-type, and mutant KCNQ2 alleles were transfected into HEK293 cells before whole-cell patch-clamp analysis. Neurodevelopmental outcomes were worst in patients with the p.R213Q mutation, better in patients with the p.E515D mutation, and best in patients with the novel p.V543 M mutation...
January 1, 2017: Journal of Child Neurology
https://www.readbyqxmd.com/read/28396143/lethal-effects-of-an-insecticidal-spider-venom-peptide-involve-positive-allosteric-modulation-of-insect-nicotinic-acetylcholine-receptors
#9
Monique J Windley, Irina Vetter, Richard J Lewis, Graham M Nicholson
κ-Hexatoxins (κ-HXTXs) are a family of excitotoxic insect-selective neurotoxins from Australian funnel-web spiders that are lethal to a wide range of insects, but display no toxicity towards vertebrates. The prototypic κ-HXTX-Hv1c selectively blocks native and expressed cockroach large-conductance calcium-activated potassium (BKCa or KCa1.1) channels, but not their mammalian orthologs. Despite this potent and selective action on insect KCa1.1 channels, we found that the classical KCa1.1 blockers paxilline, charybdotoxin and iberiotoxin, which all block insect KCa1...
April 8, 2017: Neuropharmacology
https://www.readbyqxmd.com/read/28394849/inhibition-of-voltage-gated-na-channels-by-bupivacaine-is-enhanced-by-the-adjuvants-buprenorphine-ketamine-and-clonidine
#10
Carsten Stoetzer, Christine Martell, Jeanne de la Roche, Andreas Leffler
BACKGROUND AND OBJECTIVES: Regional anesthesia includes application of local anesthetics (LAs) into the vicinity of peripheral nerves. Prolongation or improvement of nerve blocks with LAs can be accomplished by coapplication with adjuvants, including buprenorphine, ketamine, and clonidine. While the mechanisms mediating prolonged or improved LA-induced effects by adjuvants are poorly understood, we hypothesized that they are likely to increase LA-induced block of voltage-gated Na channels...
April 7, 2017: Regional Anesthesia and Pain Medicine
https://www.readbyqxmd.com/read/28394013/intermittent-theta-burst-stimulation-rescues-dopamine-dependent-corticostriatal-synaptic-plasticity-and-motor-behavior-in-experimental-parkinsonism-possible-role-of-glial-activity
#11
Fabrizio Cacace, Desirèe Mineo, Maria Teresa Viscomi, Emanuele Claudio Latagliata, Maria Mancini, Valeria Sasso, Anna Vannelli, Tiziana Pascucci, Valentina Pendolino, Elena Marcello, Silvia Pelucchi, Stefano Puglisi-Allegra, Marco Molinari, Barbara Picconi, Paolo Calabresi, Veronica Ghiglieri
BACKGROUND: Recent studies support the therapeutic utility of repetitive transcranial magnetic stimulation in Parkinson's disease (PD), whose progression is correlated with loss of corticostriatal long-term potentiation and long-term depression. Glial cell activation is also a feature of PD that is gaining increasing attention in the field because astrocytes play a role in chronic neuroinflammatory responses but are also able to manage dopamine (DA) levels. METHODS: Intermittent theta-burst stimulation protocol was applied to study the effect of therapeutic neuromodulation on striatal DA levels measured by means of in vivo microdialysis in 6-hydroxydopamine-hemilesioned rats...
April 10, 2017: Movement Disorders: Official Journal of the Movement Disorder Society
https://www.readbyqxmd.com/read/28393175/trpm7-channels-mediate-the-functional-changes-in-cardiac-fibroblasts-induced-by-angiotensin-ii
#12
Sha Li, Mingjiang Li, Xin Yi, Furong Guo, Yanli Zhou, Suqin Chen, Xian Wu
Transient receptor potential melastatin 7 (TRPM7), a bifunctional channel protein owning both cation permeability and kinase activity, plays an important role in the pathophysiological process of many cell types, such as vascular smooth muscle cells, human glioma cells and mouse cortical astrocytes. However, whether TRPM7 channels play a key role in the functional change of cardiac fibroblasts (CFs) induced by angiotensin II (Ang II) remains unknown. Using Cell Counting Kit-8 (CCK-8) assay, immunofluorescence assay, western blot analysis, RT-qPCR, RNA interference (RNAi) and whole-cell patch-clamp techniques, the present study aimed to explore the role of TRPM7 channels in the proliferation, differentiation and collagen synthesis of CFs induced by Ang II...
April 6, 2017: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/28389870/orexin-directly-enhances-the-excitability-of-globus-pallidus-internus-neurons-in-rat-by-co-activating-ox1-and-ox2-receptors
#13
He-Ren Gao, Qian-Xing Zhuang, Yong-Xiao Zhang, Zhang-Peng Chen, Bin Li, Xiao-Yang Zhang, Yi-Ting Zhong, Jian-Jun Wang, Jing-Ning Zhu
Orexin, released from the hypothalamus, has been implicated in various basic non-somatic functions including feeding, the sleep-wakefulness cycle, emotion, and cognition. However, the role of orexin in somatic motor control is still little known. Here, using whole-cell patch clamp recording and immunostaining, we investigated the effect and the underlying receptor mechanism of orexin-A on neurons in the globus pallidus internus (GPi), a critical structure in the basal ganglia and an effective target for deep brain stimulation therapy...
April 7, 2017: Neuroscience Bulletin
https://www.readbyqxmd.com/read/28388364/piezo2-expression-in-mechanosensitive-dental-primary-afferent-neurons
#14
J Won, H Vang, P R Lee, Y H Kim, H W Kim, Y Kang, S B Oh
Mechanosensitive ion channels have been suggested to be expressed in dental primary afferent (DPA) neurons to transduce the movement of dentinal fluid since the proposal of hydrodynamic theory. Piezo2, a mechanosensitive, rapidly inactivating (RI) ion channel, has been recently identified in dorsal root ganglion (DRG) neurons to mediate tactile transduction. Here, we examined the expression of Piezo2 in DPA neurons by in situ hybridization, single-cell reverse transcriptase polymerase chain reaction, and whole-cell patch-clamp recordings...
April 1, 2017: Journal of Dental Research
https://www.readbyqxmd.com/read/28385341/enhancement-of-%C3%AE-opioid-receptor-desensitization-by-orexin-a-in-rat-locus-coeruleus-neurons
#15
S Mohammad Ahmadi Soleimani, Hossein Azizi, Narges Pachenari, Javad Mirnajafi-Zadeh, Saeed Semnanian
Opioids have always been used in clinical practice for pain management. However, development of tolerance to their effects following long term administration, seriously restricts further clinical use of these drugs. In this regard, μ-opioid receptor (MOR) desensitization, as an initial step in development of opioid tolerance, is of particular significance. Previous studies support the involvement of orexinergic system in development of opioid tolerance. Locus coeruleus (LC) nucleus has been shown to modulate pain and development of tolerance...
March 23, 2017: Neuropeptides
https://www.readbyqxmd.com/read/28380698/myoclonus-epilepsy-and-ataxia-due-to-kcnc1-mutation-analysis-of-20-cases-and-k-channel-properties
#16
Karen L Oliver, Silvana Franceschetti, Carol J Milligan, Mikko Muona, Simone A Mandelstam, Laura Canafoglia, Anna M Boguszewska-Chachulska, Amos Korczyn, Francesca Bisulli, Carlo Di Bonaventura, Francesca Ragona, Roberto Michelucci, Bruria Ben-Zeev, Rachel Straussberg, Ferruccio Panzica, João Massano, Daniel Friedman, Arielle Crespel, Bernt A Engelsen, Frederick Andermann, Eva Andermann, Krystyna Spodar, Anetta Lasek-Bal, Patrizia Riguzzi, Elena Pasini, Paolo Tinuper, Laura Licchetta, Elena Gardella, Matthias Lindenau, Annette Wulf, Rikke S Møller, Felix Benninger, Zaid Afawi, Guido Rubboli, Christopher A Reid, Snezana Maljevic, Holger Lerche, Anna-Elina Lehesjoki, Steven Petrou, Samuel F Berkovic
OBJECTIVE: To comprehensively describe the new syndrome of myoclonus epilepsy and ataxia due to potassium (K(+) ) channel mutation (MEAK), including cellular electrophysiological characterization of observed clinical improvement with fever. METHODS: We analysed clinical, electroclinical and neuroimaging data for twenty patients with MEAK due to recurrent KCNC1 p.R320H mutation. In vitro electrophysiological studies were conducted using whole cell patch clamp to explore biophysical properties of wild-type and mutant KV 3...
April 5, 2017: Annals of Neurology
https://www.readbyqxmd.com/read/28379373/genetic-and-phenotypic-heterogeneity-suggest-therapeutic-implications-in-scn2a-related-disorders
#17
Markus Wolff, Katrine M Johannesen, Ulrike B S Hedrich, Silvia Masnada, Guido Rubboli, Elena Gardella, Gaetan Lesca, Dorothée Ville, Mathieu Milh, Laurent Villard, Alexandra Afenjar, Sandra Chantot-Bastaraud, Cyril Mignot, Caroline Lardennois, Caroline Nava, Niklas Schwarz, Marion Gérard, Laurence Perrin, Diane Doummar, Stéphane Auvin, Maria J Miranda, Maja Hempel, Eva Brilstra, Nine Knoers, Nienke Verbeek, Marjan van Kempen, Kees P Braun, Grazia Mancini, Saskia Biskup, Konstanze Hörtnagel, Miriam Döcker, Thomas Bast, Tobias Loddenkemper, Lily Wong-Kisiel, Friedrich M Baumeister, Walid Fazeli, Pasquale Striano, Robertino Dilena, Elena Fontana, Federico Zara, Gerhard Kurlemann, Joerg Klepper, Jess G Thoene, Daniel H Arndt, Nicolas Deconinck, Thomas Schmitt-Mechelke, Oliver Maier, Hiltrud Muhle, Beverly Wical, Claudio Finetti, Reinhard Brückner, Joachim Pietz, Günther Golla, Dinesh Jillella, Karen M Linnet, Perrine Charles, Ute Moog, Eve Õiglane-Shlik, John F Mantovani, Kristen Park, Marie Deprez, Damien Lederer, Sandrine Mary, Emmanuel Scalais, Laila Selim, Rudy Van Coster, Lieven Lagae, Marina Nikanorova, Helle Hjalgrim, G Christoph Korenke, Marina Trivisano, Nicola Specchio, Berten Ceulemans, Thomas Dorn, Katherine L Helbig, Katia Hardies, Hannah Stamberger, Peter de Jonghe, Sarah Weckhuysen, Johannes R Lemke, Ingeborg Krägeloh-Mann, Ingo Helbig, Gerhard Kluger, Holger Lerche, Rikke S Møller
Mutations in SCN2A, a gene encoding the voltage-gated sodium channel Nav1.2, have been associated with a spectrum of epilepsies and neurodevelopmental disorders. Here, we report the phenotypes of 71 patients and review 130 previously reported patients. We found that (i) encephalopathies with infantile/childhood onset epilepsies (≥3 months of age) occur almost as often as those with an early infantile onset (<3 months), and are thus more frequent than previously reported; (ii) distinct phenotypes can be seen within the late onset group, including myoclonic-atonic epilepsy (two patients), Lennox-Gastaut not emerging from West syndrome (two patients), and focal epilepsies with an electrical status epilepticus during slow sleep-like EEG pattern (six patients); and (iii) West syndrome constitutes a common phenotype with a major recurring mutation (p...
March 4, 2017: Brain: a Journal of Neurology
https://www.readbyqxmd.com/read/28377087/monocyte-derived-macrophages-from-cf-pigs-exhibit-increased-inflammatory-responses-at-birth
#18
Lily Paemka, Brian N McCullagh, Mahmoud H Abou Alaiwa, David A Stoltz, Qian Dong, Christoph O Randak, Robert D Gray, Paul B McCray
BACKGROUND: We sought to address whether CF macrophages have a primary functional defect as a consequence of CFTR loss and thus contribute to the onset of infection and inflammation observed in CF lung disease. METHODS: Monocyte derived macrophages (MDMs) were prepared from newborn CF and non-CF pigs. CFTR mRNA expression was quantified by rtPCR and anion channel function was determined using whole cell patch clamp analysis. IL8 and TNFα release from MDMs in response to lipopolysaccharide stimulation was measured by ELISA...
April 1, 2017: Journal of Cystic Fibrosis: Official Journal of the European Cystic Fibrosis Society
https://www.readbyqxmd.com/read/28373496/in-vivo-patch-clamp-recording-in-awake-head-fixed-rodents
#19
Doyun Lee, Albert K Lee
Whole-cell recording has been used to measure and manipulate a neuron's spiking and subthreshold membrane potential, allowing assessment of the cell's inputs and outputs as well as its intrinsic membrane properties. This technique has also been combined with pharmacology and optogenetics as well as morphological reconstruction to address critical questions concerning neuronal integration, plasticity, and connectivity. This protocol describes a technique for obtaining whole-cell recordings in awake head-fixed animals, allowing such questions to be investigated within the context of an intact network and natural behavioral states...
April 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28373245/an-east-asian-common-variant-vinculin-p-asp841his-was-associated-with-sudden-unexplained-nocturnal-death-syndrome-in-the-chinese-han-population
#20
Jianding Cheng, John W Kyle, Di Lang, Brandi Wiedmeyer, Jian Guo, Kun Yin, Lei Huang, Ravi Vaidyanathan, Terry Su, Jonathan C Makielski
BACKGROUND: We have identified the cardiomyopathy-susceptibility gene vinculin (VCL) mutation M94I may account for a sudden unexplained nocturnal death syndrome (SUNDS) case. We addressed whether VCL common variant D841H is associated with SUNDS. METHODS AND RESULTS: In 8 of 120 SUNDS cases, we detected an East Asian common VCL variant p.Asp841His (D841H). Comparing the H841 allele frequency of the general population in the local database (15 of 1818) with SUNDS victims (10 of 240) gives an odds ratio for SUNDS of 5...
April 3, 2017: Journal of the American Heart Association
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