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Whole cell patch clamp

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https://www.readbyqxmd.com/read/28533887/lidocaine-suppresses-glioma-cell-proliferation-by-inhibiting-trpm7-channels
#1
Tiandong Leng, Suizhen Lin, Zhigang Xiong, Jun Lin
BACKGROUND: Malignant glioma is the most common brain cancer with devastating prognosis. Recurrence of malignant glioma following surgery is very common with few preventive and therapeutic options. Novel targets and therapeutic agents are constantly sought for better outcome. Our previous study established that inhibition of transient receptor potential melastatin 7 (TRPM7) channels resulted in significant decrease of human glioma cell growth and proliferation. As local anesthetic lidocaine has been shown to inhibit TRPM7 currents, we hypothesize that lidocaine may suppress glioma cell proliferation through TRPM7 channel inhibition...
2017: International Journal of Physiology, Pathophysiology and Pharmacology
https://www.readbyqxmd.com/read/28533740/correlation-of-ventricular-arrhythmogenesis-with-neuronal-remodeling-of-cardiac-postganglionic-parasympathetic-neurons-in-the-late-stage-of-heart-failure-after-myocardial-infarction
#2
Dongze Zhang, Huiyin Tu, Chaojun Wang, Liang Cao, Robert L Muelleman, Michael C Wadman, Yu-Long Li
Introduction: Ventricular arrhythmia is a major cause of sudden cardiac death in patients with chronic heart failure (CHF). Our recent study demonstrates that N-type Ca(2+) currents in intracardiac ganglionic neurons are reduced in the late stage of CHF rats. Rat intracardiac ganglia are divided into the atrioventricular ganglion (AVG) and sinoatrial ganglion. Only AVG nerve terminals innervate the ventricular myocardium. In this study, we tested the correlation of electrical remodeling in AVG neurons with ventricular arrhythmogenesis in CHF rats...
2017: Frontiers in Neuroscience
https://www.readbyqxmd.com/read/28533091/na-h-exchange-regulatory-factor-1-is-required-for-romk1-k-channel-expression-in-the-surface-membrane-of-cultured-m-1-cortical-collecting-duct-cells
#3
Takashi Suzuki, Kazuyoshi Nakamura, Taira Mayanagi, Kenji Sobue, Manabu Kubokawa
The ROMK1 K(+) channel, a member of the ROMK channel family, is the major candidate for the K(+) secretion pathway in the renal cortical collecting duct (CCD). ROMK1 possesses a PDZ domain-binding motif at its C-terminus that is considered a modulator of ROMK1 expression via interaction with Na(+)/H(+) exchange regulatory factor (NHERF) 1 and NHERF2 scaffold protein. Although NHERF1 is a potential binding partner of the ROMK1 K(+) channel, the interaction between NHERF1 and K(+) channel activity remains unclear...
May 19, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28522805/nitric-oxide-donor-protects-against-acetic-acid-induced-gastric-ulcer-in-rats-via-s-nitrosylation-of-trpv1-on-vagus-nerve
#4
Ting Han, Yan Tang, Jing Li, Bing Xue, Liping Gong, Jingxin Li, Xiao Yu, Chuanyong Liu
This study was conducted to investigate the effects of nitric oxide (NO) in acetic acid-induced gastric ulcer of rats and the underlying mechanisms. We found that peritoneal injection of sodium nitroprusside (SNP), a NO donor, decreased the ulcer area, inflammatory cell infiltration and MPO degree in acetic acid-induced gastric ulcer in rats. This effect was abolished by a transient receptor potential vanilloid 1 (TRPV1) antagonist or prior subdiaphragmatic vagotomy. SNP increased the jejunal mesenteric afferent discharge in a dose-depended manner, which was largely diminished by pretreatment of S-nitrosylation blocker N-ethylmaleimide, TRPV1 antagonist capsazepine, genetic deletion of TRPV1, or vagotomy...
May 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28520841/chronic-inflammatory-pain-impairs-mglur5-mediated-depolarization-induced-suppression-of-excitation-in-the-anterior-cingulate-cortex
#5
Baolin Guo, Jiaqi Wang, Han Yao, Keke Ren, Jing Chen, Jing Yang, Guohong Cai, Haiying Liu, Yunlong Fan, Wenting Wang, Shengxi Wu
The anterior cingulate cortex (ACC) is a critical hub for nociceptive perception and pain-related anxiety. Long-term synaptic plasticity in ACC was found to be important for chronic inflammatory pain and pain-related anxiety. As short-term synaptic plasticity, depolarization-induced suppression of excitation (DSE) is involved in several conditions, such as chronic stress, epilepsy, and autism. However, it is still unknown whether DSE in the ACC is involved in the central sensitization of pain and anxiety. Using a whole-cell patch clamp, calcium imaging, western blot, and behavioral testing, we found that DSE was induced by a 2 s depolarization in postsynaptic pyramidal cells in ACC...
May 17, 2017: Cerebral Cortex
https://www.readbyqxmd.com/read/28514017/isopimaric-acid-a-multi-targeting-ion-channel-modulator-reducing-excitability-and-arrhythmicity-in-a-spontaneously-beating-mouse-atrial-cell-line
#6
Sajjad Salari, Malin Silverå Ejneby, Johan Brask, Fredrik Elinder
AIM: Atrial fibrillation is the most common persistent cardiac arrhythmia, and it is not well controlled by present drugs. Because some resin acids open voltage-gated potassium channels and reduce neuronal excitability, we explored the effects of the resin acid isopimaric acid (IPA) on action potentials and ion currents in cardiomyocytes. METHODS: Spontaneously beating mouse atrial HL-1 cells were investigated with the whole-cell patch-clamp technique. RESULTS: 1-25 μmol L(-1) IPA reduced the action potential frequency by up to 50%...
May 17, 2017: Acta Physiologica
https://www.readbyqxmd.com/read/28513236/enhanced-gap-junctional-channel-activity-between-vascular-smooth-muscle-cells-in-cerebral-artery-of-spontaneously-hypertensive-rats
#7
Li-Jie Wang, Ke-Tao Ma, Wen-Yan Shi, Ying-Zi Wang, Lei Zhao, Xin-Yan Chen, Xin-Zhi Li, Xue-Wei Jiang, Zhong-Shuang Zhang, Li Li, Jun-Qiang Si
The aim of the present study is to investigate the effects of hypertension on the gap junctions between vascular smooth muscle cells (VSMCs) in the cerebral arteries (CAs) of spontaneously hypertensive rats (SHRs). The functions of gap junctions in the CAs of VSMCs in SHRs and control normotensive Wistar-Kyoto (WKY) rats were studied using whole-cell patch clamp recordings and pressure myography, and the expression levels of connexins were analyzed using reverse transcription-quantitative polymerase chain reaction and Western blot analyses...
May 17, 2017: Clinical and Experimental Hypertension: CHE
https://www.readbyqxmd.com/read/28512142/a-well-known-potassium-channel-plays-a-critical-role-in-lysosomes
#8
Michael X Zhu
Whole-endolysosome patch clamping presents new opportunities to identify and characterize channels pivotal for these acidic organelles. In this issue (Wang et al., 2017. J. Cell Biol. https://doi.org/10.1083/jcb.201612123), the identification of a role for the large conductance calcium-activated potassium channel brings new thinking about regulation of lysosome membrane potential and function.
May 16, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28511870/atp-sensitive-k-channels-maintain-resting-membrane-potential-in-interstitial-cells-of-cajal-from-the-mouse-colon
#9
Ji Sun Na, Chansik Hong, Man Woo Kim, Chan Guk Park, Hyun Goo Kang, Mei Jin Wu, Han Yi Jiao, Seok Choi, Jae Yeoul Jun
To investigate the role of ATP-sensitive K(+) (KATP) channels on pacemaker activity in interstitial cells of Cajal (ICC), whole-cell patch clamping, RT-PCR, and intracellular Ca(2+) ([Ca(2+)]i) imaging were performed in cultured colonic ICC. Pinacidil (a K(+) channel opener) hyperpolarized the membrane and inhibited the generation of pacemaker potential, and this effect was reversed by glibenclamide (a KATP channel blocker). RT-PCR showed that Kir 6.1 and SUR2B were expressed in Ano-1 positive colonic ICC. Glibenclamide depolarized the membrane and increased pacemaker potential frequency...
May 13, 2017: European Journal of Pharmacology
https://www.readbyqxmd.com/read/28508747/voltage-gated-na-currents-in-human-dorsal-root-ganglion-neurons
#10
Xiulin Zhang, Birgit T Priest, Inna Belfer, Michael S Gold
Available evidence indicates voltage-gated Na(+) channels (VGSCs) in peripheral sensory neurons are essential for the pain and hypersensitivity associated with tissue injury. However, our understanding of the biophysical and pharmacological properties of the channels in sensory neurons is largely based on the study of heterologous systems or rodent tissue, despite evidence that both expression systems and species differences influence these properties. Therefore, we sought to determine the extent to which the biophysical and pharmacological properties of VGSCs were comparable in rat and human sensory neurons...
May 16, 2017: ELife
https://www.readbyqxmd.com/read/28499969/corticotropin-releasing-hormone-modulates-airway-vagal-preganglionic-neurons-of-sprague-dawley-rats-at-multiple-synaptic-sites-via-activation-of-its-type-1-receptors-implications-for-stress-associated-airway-vagal-excitation
#11
Yuhong Guo, Xianxia Yan, Xingxin Chen, Ding He, Ming Zeng, Yonghua Chen, Chunmei Xia, Dongying Qiu, Jijiang Wang
Corticotropin-releasing hormone release is the final common pathway of stress-associated neuroendocrine responses. This study tested how corticotropin-releasing hormone modulates airway vagal preganglionic neurons. Airway vagal preganglionic neurons in neonatal rats were retrogradely labeled with fluorescent dye and identified in medullary slices, and their responses to corticotropin-releasing hormone (200nmolL(-1)) were examined using whole-cell patch clamp. The results show that under current clamp, corticotropin-releasing hormone (200nmolL(-1)) depolarized airway vagal preganglionic neurons and significantly increased the rate of their spontaneous firing...
May 10, 2017: Neuroscience
https://www.readbyqxmd.com/read/28498410/neuropeptide-y-suppresses-epileptiform-discharges-by-regulating-ampa-receptor-glur2-subunit-in-rat-hippocampal-neurons
#12
Wei Bu, Wen-Qing Zhao, Wen-Ling Li, Chang-Zheng Dong, Zhe Zhang, Qi-Jun Li
The present study aimed to investigate the effects of neuropeptide Y (NPY) on the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor glutamate receptor 2 (GluR2) subunit in epileptiform discharge hippocampal neurons. Hippocampal neurons were harvested from neonatal Sprague‑Dawley rats aged <24 h and primarily cultured in vitro. At day 12 following culture, hippocampal neurons were divided into the following groups: Control, Mg2+‑free, NPY+Mg2+‑free and BIBP3226+NPY+Mg2+‑free...
May 10, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28489663/use-dependent-inhibition-of-glycine-activated-chloride-current-in-rat-neurons-by-%C3%AE-amyloid-peptide-pretreated-with-hexafluoroisopropanol
#13
Elena I Solntseva, Julia V Bukanova
Hexafluoroisopropanol (HFIP) is a nonpolar organic solvent that is often used to prepare β-amyloid peptide (Aβ) samples. In this work, we compare the effects of two different species derived from synthetic Aβ1-42 and prepared without HFIP (Aβ) or using HFIP (Aβ/HFIP) on the glycine-activated chloride current (IGly). The experiments were conducted on the pyramidal neurons isolated from CA3 region of rat hippocampus. Transmembrane currents were recorded using a conventional patch-clamp technique in the whole-cell configuration...
May 5, 2017: Neuroreport
https://www.readbyqxmd.com/read/28487481/functional-genomics-in-brugia-malayi-reveal-diverse-muscle-nachrs-and-differences-between-cholinergic-anthelmintics
#14
Saurabh Verma, Sudhanva Srinivas Kashyap, Alan Patrick Robertson, Richard John Martin
Many techniques for studying functional genomics of important target sites of anthelmintics have been restricted to Caenorhabditis elegans because they have failed when applied to animal parasites. To overcome these limitations, we have focused our research on the human nematode parasite Brugia malayi, which causes elephantiasis. Here, we combine single-cell PCR, whole muscle cell patch clamp, motility phenotyping (Worminator), and dsRNA for RNAi for functional genomic studies that have revealed, in vivo, four different muscle nAChRs (M-, L-, P-, and N-)...
May 9, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28487470/discrepant-effects-of-heart-failure-on-electrophysiological%C3%A2-property-in-right-ventricular-outflow-tract-and-left-ventricular-outflow-tract-cardiomyocytes
#15
Yen-Yu Lu, Chen-Chuan Cheng, Chin-Feng Tsai, Yung Kuo Lin, Ting-I Lee, Yao-Chang Chen, Shih-Ann Chen, Yi Jen Chen
Ventricular arrhythmias commonly arise from the right and left ventricular outflow tracts (RVOT/LVOT) in patients without structural heart disease. Heart failure (HF) significantly increases the risk of ventricular arrhythmias. The regional differences and how HF affects the electrophysiological characteristics of RVOT and LVOT cardiomyocytes remain unclear. The whole-cell patch clamp technique was used to investigate the action potentials and ionic currents in isolated single RVOT and LVOT cardiomyocytes from control and rapid ventricular pacing-induced HF rabbit...
May 9, 2017: Clinical Science (1979-)
https://www.readbyqxmd.com/read/28483393/activation-of-5-ht7-receptors-reverses-nmda-r-dependent-ltd-by-activating-pka-in-medial-vestibular-neurons
#16
Yan-Hai Li, Lei Han, Kenneth Lap-Kei Wu, Ying-Shing Chan
The medial vestibular nucleus (MVN) is a major output station for neurons that project to the vestibulo-spinal pathway. MVN neurons show capacity for long-term depression (LTD) during the juvenile period. We investigated LTD of MVN neurons using whole-cell patch-clamp recordings. High frequency stimulation (HFS) robustly induced LTD in 90% of type B neurons in the MVN, while only 10% of type A neurons were responsive, indicating that type B neurons are the major contributors to LTD in the MVN. The neuromodulator serotonin (5-HT) is known to modulate LTD in neural circuits of the cerebral cortex and the hippocampus...
May 5, 2017: Neuropharmacology
https://www.readbyqxmd.com/read/28476169/effects-of-diacetyl-liensinine-on-electrophysiology-in-rabbit-ventricular-myocytes
#17
Feng Cao, Teng Wang, Wenmao Ding, Zhe Li, Shaobo Shi, Xiaozhan Wang
BACKGROUND: Diacetyl-liensinine is a chemosynthetic derivative of liensinine, extracted from the seed embryo of Nelumbo nucifera Gaertn, in China. It has been found to have extensive anti- arrhythmic actions. The present study was designed to investigate the effects of diacetyl-liensinine on electro- physiology of myocytes. METHODS: We exposed rabbit ventricular myocytes to diacetyl-liensinine using standard whole-cell patch-clamp technique and measured the action potential, L-type calcium current (I Ca-L), delayed rectifier potassium current (I K), transient outward potassium current (I to) and inward rectifier potassium current (I K1)...
May 5, 2017: BMC Pharmacology & Toxicology
https://www.readbyqxmd.com/read/28473457/pharmacologic-characterization-of-amg8379-a-potent-and-selective-small-molecule-sulfonamide-antagonist-of-the-voltage-gated-sodium-channel-nav1-7
#18
Thomas J Kornecook, Ruoyuan Yin, Stephen Altmann, Xuhai Be, Virginia Berry, Christopher P Ilch, Michael Jarosh, Danielle Johnson, Josie H Lee, Sonya G Lehto, Joseph Ligutti, Dong Liu, Jason Luther, David Matson, Danny Ortuno, John Roberts, Kristin Taborn, Jinti Wang, Matthew M Weiss, Violeta Yu, Dawn X D Zhu, Robert T Fremeau, Bryan D Moyer
Potent and selective antagonists of the voltage-gated sodium channel NaV1.7 represent a promising avenue for the development of new chronic pain therapies. We generated a small molecule atropisomer quinolone sulfonamide antagonist AMG8379 and a less active enantiomer AMG8380. Here we show that AMG8379 potently blocks human NaV1.7 channels with an IC50 of 8.5 nM and endogenous tetrodotoxin (TTX)-sensitive sodium channels in dorsal root ganglia (DRG) neurons with an IC50 of 3.1 nM in whole cell patch clamp electrophysiology assays using a voltage protocol that interrogates channels in a partially inactivated state...
May 4, 2017: Journal of Pharmacology and Experimental Therapeutics
https://www.readbyqxmd.com/read/28472842/diterpene-lipo-alkaloids-with-selective-activities-on-cardiac-k-channels
#19
Tivadar Kiss, Botond Borcsa, Péter Orvos, László Tálosi, Judit Hohmann, Dezső Csupor
Aconitum diterpene alkaloids are known for their remarkable toxicity, which is due to their effect on ion channels. Activation of voltage-gated Na(+) channels is the major cause of their cardiotoxicity, however, influence on K(+) channels may also play a role in the overall effect.Here we report the synthesis of a series of lipo-alkaloids, including four new compounds, based on the 14-benzoylaconine structure, which is the core of a vast number of diterpene alkaloids naturally occurring in Aconitum species...
May 4, 2017: Planta Medica
https://www.readbyqxmd.com/read/28472301/new-gain-of-function-mutation-shows-cacna1d-as-recurrently-mutated-gene-in-autism-spectrum-disorders-and-epilepsy
#20
Alexandra Pinggera, Luisa Mackenroth, Andreas Rump, Jens Schallner, Filippo Beleggia, Bernd Wollnik, Jörg Striessnig
CACNA1D encodes the pore-forming α1-subunit of Cav1.3, an L-type voltage-gated Ca2+-channel. Despite the recent discovery of two de novo missense gain-of-function mutations in Cav1.3 in two individuals with autism spectrum disorder (ASD) and intellectual disability CACNA1D has not been considered a prominent ASD-risk gene in large scale genetic analyses, since such studies primarly focus on likely-disruptive genetic variants. Here we report the discovery and characterization of a third de novo missense mutation in CACNA1D (V401L) in a patient with ASD and epilepsy...
May 4, 2017: Human Molecular Genetics
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