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Whole cell patch clamp

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https://www.readbyqxmd.com/read/28943120/diversity-of-bilateral-synaptic-assemblies-for-binaural-computation-in-midbrain-single-neurons
#1
Na He, Lingzhi Kong, Tao Lin, Shaohui Wang, Xiuping Liu, Jiyao Qi, Jun Yan
Binaural hearing confers many beneficial functions but our understanding of its underlying neural substrates is limited. This study examines the bilateral synaptic assemblies and binaural computation (or integration) in the central nucleus of the inferior colliculus (ICc) of the auditory midbrain, a key convergent center. Using in-vivo whole-cell patch-clamp, the excitatory and inhibitory postsynaptic potentials (EPSPs/IPSPs) of single ICc neurons to contralateral, ipsilateral and bilateral stimulation were recorded...
September 18, 2017: Hearing Research
https://www.readbyqxmd.com/read/28941256/distinct-protein-kinase-c-isoenzyme-mediates-inhibitory-effects-of-different-g-protein-coupled-receptors-on-cardiac-rapidly-activating-delayed-rectifier-k-current
#2
Xueli Liu, Yuhong Wang, Hua Zhang, Li Shen, Yanfang Xu
BACKGROUND AND PURPOSE: Elevated angiotensin II (Ang II) and sympathetic activity contributes to high risk of ventricular arrhythmias in heart diseases. The rapidly activating delayed rectifier K(+) current (IKr ) carried by the hERG protein plays a critical role in cardiac repolarization; and a reduction of IKr is involved in increased cardiac arrhythmogenicity. Studies have shown that stimulation of α1A -adrenoreceptors (α1A AR) or Ang II type 1 receptor (AT1 R) inhibits IKr via protein kinase C (PKC)...
September 23, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28940424/n1366s-mutation-of-human-skeletal-muscle-sodium-channel-causes-paramyotonia-congenita
#3
Qing Ke, Jia Ye, Siyang Tang, Jin Wang, Benyan Luo, Fang Ji, Xu Zhang, Ye Yu, Xiaoyang Cheng, Yuezhou Li
Paramyotonia congenita is an autosomal dominant skeletal muscle channelopathy caused by missense mutations in SCN4A, the gene encoding the α subunit of the human skeletal muscle voltage-gated sodium channel NaV1.4. Here, we report a three-generation family in which six members present clinical symptoms of paramyotonia congenita characterized by a marked worsening of myotonia by cold and by the presence of clear episodes of paralysis. We identified a novel mutation in SCN4A (Asn-1366-Ser, N1366S) in all patients in the family but not in healthy relatives or in 500 normal control subjects...
September 20, 2017: Journal of Physiology
https://www.readbyqxmd.com/read/28939258/novel-calcium-related-targets-of-insulin-in-hippocampal-neurons
#4
Shaniya Maimaiti, Hilaree N Frazier, Katie L Anderson, Adam O Ghoweri, Lawrence D Brewer, Nada M Porter, Olivier Thibault
Both insulin signaling disruption and Ca(2+) dysregulation are closely related to memory loss during aging and increase the vulnerability to Alzheimer's disease (AD). In hippocampal neurons, aging-related changes in calcium regulatory pathways have been shown to lead to higher intracellular calcium levels and an increase in the Ca(2+)-dependent afterhyperpolarization (AHP), which is associated with cognitive decline. Recent studies suggest that insulin reduces the Ca(2+)-dependent AHP. Given the sensitivity of neurons to insulin and evidence that brain insulin signaling is reduced with age, insulin-mediated alterations in calcium homeostasis may underlie the beneficial actions of insulin in the brain...
September 19, 2017: Neuroscience
https://www.readbyqxmd.com/read/28935562/n-2-methoxyphenyl-benzenesulfonamide-a-novel-regulator-of-neuronal-g-protein-gated-inward-rectifier-k-channels
#5
Kenneth B Walsh, Elaine A Gay, Bruce E Blough, David W Geurkink
G protein-gated inward rectifier K(+) (GIRK) channels are members of the super-family of proteins known as inward rectifier K(+) (Kir) channels and are expressed throughout the peripheral and central nervous systems. Neuronal GIRK channels are the downstream targets of a number of neuromodulators including opioids, somatostatin, dopamine and cannabinoids. Previous studies have demonstrated that the ATP-sensitive K(+) channel, another member of the Kir channel family, is regulated by sulfonamide drugs. Therefore, to determine if sulfonamides also modulate GIRK channels, we screened a library of arylsulfonamide compounds using a GIRK channel fluorescent assay that utilized pituitary AtT20 cells expressing GIRK channels along with the somatostatin type-2 and -5 receptors...
September 18, 2017: European Journal of Pharmacology
https://www.readbyqxmd.com/read/28931666/trpa1-activation-in-a-human-sensory-neuronal-model-relevance-to-cough-hypersensitivity
#6
Rebecca Clarke, Kevin Monaghan, Imad About, Caoimhin S Griffin, Gerard P Sergeant, Ikhlas El Karim, J Graham McGeown, S Louise Cosby, Timothy M Curtis, Lorcan P McGarvey, Fionnuala T Lundy
The cough reflex becomes hyperresponsive in acute and chronic respiratory diseases, but understanding the underlying mechanism is hampered by difficulty accessing human tissue containing both nerve endings and neuronal cell bodies. We refined an adult stem cell sensory neuronal model to overcome the limited availability of human neurones and applied the model to study transient receptor potential ankyrin 1 (TRPA1) channel expression and activation.Human dental pulp stem cells (hDPSCs) were differentiated towards a neuronal phenotype, termed peripheral neuronal equivalents (PNEs)...
September 2017: European Respiratory Journal: Official Journal of the European Society for Clinical Respiratory Physiology
https://www.readbyqxmd.com/read/28931571/sublaminar-subdivision-of-mouse-auditory-cortex-layer-2-3-based-on-functional-translaminar-connections
#7
Xiangying Meng, Daniel E Winkowski, Joseph P Y Kao, Patrick O Kanold
The cerebral cortex is subdivided into 6 layers based on morphological features. The supragranular layers 2/3 (L2/3) contain morphologically and genetically diverse populations of neurons, suggesting the existence of discrete classes of cells. In primates and carnivores L2/3 can be subdivided morphologically, but cytoarchitectonic divisions are less clear in rodents. Nevertheless, discrete classes of cells could exist based on their computational requirement, which might be linked to their associated functional microcircuits...
September 20, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/28927958/defective-gabaergic-neurotransmission-in-the-nucleus-tractus-solitarius-in-mecp2-null-mice-a-model-of-rett-syndrome
#8
Chao-Yin Chen, Jacopo Di Lucente, Yen-Chu Lin, Cheng-Chang Lien, Michael A Rogawski, Izumi Maezawa, Lee-Way Jin
Rett syndrome (RTT) is a devastating neurodevelopmental disorder caused by loss-of-function mutations in the X-linked methyl-CpG binding protein 2 (Mecp2) gene. GABAergic dysfunction has been implicated contributing to the respiratory dysfunction, one major clinical feature of RTT. The nucleus tractus solitarius (NTS) is the first central site integrating respiratory sensory information that can change the nature of the reflex output. We hypothesized that deficiency in Mecp2 gene reduces GABAergic neurotransmission in the NTS...
September 16, 2017: Neurobiology of Disease
https://www.readbyqxmd.com/read/28926472/differential-sensitivity-of-presynaptic-and-postsynaptic-gabab-receptors-in-rat-ventrolateral-periaqueductal-gray
#9
Qian Chen, Caifeng Shao, Hui Zhou, Rui Ma, Ping Jiang, Kun Yang
The ventrolateral periaqueductal gray (PAG) in the midbrain plays a key role in the descending pain modulatory system. γ-Aminobutyric acid type B (GABAB) receptors belong to a metabotropic receptor subfamily and mediate both presynaptic and postsynaptic effects in PAG. It has been well documented that activation of GABAB receptors yields analgesia in some PAG subdivisions. In the present study, employing whole-cell patch-clamp recordings on acute rat PAG slices, we simultaneously monitored the responses of presynaptic and postsynaptic GABAB receptors...
September 18, 2017: Neuroreport
https://www.readbyqxmd.com/read/28924048/a-mutant-of-the-bmk-antitumor-analgesic-peptide-exhibits-reduced-inhibition-to-hnav1-4-and-hnav1-5-channels-while-retaining-analgesic-activity
#10
Yijia Xu, Xiangxue Meng, Xue Hou, Jianfang Sun, Xiaohua Kong, Yuqi Sun, Zeyu Liu, Yuanyuan Ma, Ye Niu, Yongbo Song, Yong Cui, Mingyi Zhao, Jinghai Zhang
Scorpion toxins can kill other animals by inducing paralysis and arrhythmia, which limits the potential applications of these agents in the clinical management of diseases. Antitumor-analgesic peptide (AGAP), purified from Buthus martensii Karsch (BmK), has been proved to possess analgesic and antitumor activities. Trp38, a conserved aromatic residue of AGAP, might play an important role in mediating AGAP activities according to the sequence and homology modeling analyses. Therefore, an AGAP mutant W38G was generated, and effects of both AGAP and the mutant W38G were examine by whole-cell patch clamp techniques on the sodium channels hNav1...
September 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28916636/electrophysiological-heterogeneity-of-pacemaker-cells-in-rabbit-intercaval-region-including-sa-node-insights-from-recording-multiple-ion-currents-in-each-cell
#11
Oliver Monfredi, Kenta Tsutsui, Bruce D Ziman, Michael D Stern, Edward G Lakatta, Victor A Maltsev
Cardiac pacemaker cells, including cells of the sinoatrial node, are heterogeneous in size, morphology and electrophysiological characteristics. The exact extent to which these cells differ electrophysiologically is unclear, yet is critical to understanding their functioning. We examined major ionic currents in individual intercaval pacemaker cells (IPC) sampled from the para-cristal, inter-caval region (including the sinoatrial node) that were spontaneously beating after enzymatic isolation from rabbit hearts...
September 15, 2017: American Journal of Physiology. Heart and Circulatory Physiology
https://www.readbyqxmd.com/read/28913625/calcium-release-dependent-inactivation-precedes-formation-of-the-tubular-system-in-developing-rat-cardiac-myocytes
#12
Katarina Macková, Alexandra Zahradníková, Matej Hoťka, Barbora Hoffmannová, Ivan Zahradník, Alexandra Zahradníková
Developing cardiac myocytes undergo substantial structural and functional changes transforming the mechanism of excitation-contraction coupling from the embryonic form, based on calcium influx through sarcolemmal DHPR calcium channels, to the adult form, relying on local calcium release through RYR calcium channels of sarcoplasmic reticulum stimulated by calcium influx. We characterized day-by-day the postnatal development of the structure of sarcolemma, using techniques of confocal fluorescence microscopy, and the development of the calcium current, measured by the whole-cell patch-clamp in isolated rat ventricular myocytes...
September 14, 2017: European Biophysics Journal: EBJ
https://www.readbyqxmd.com/read/28910617/whole-cell-recording-of-neuronal-membrane-potential-during-behavior
#13
REVIEW
Carl C H Petersen
Neuronal membrane potential is of fundamental importance for the mechanistic understanding of brain function. This review discusses progress in whole-cell patch-clamp recordings for low-noise measurement of neuronal membrane potential in awake behaving animals. Whole-cell recordings can be combined with two-photon microscopy to target fluorescently labeled neurons, revealing cell-type-specific membrane potential dynamics of retrogradely or genetically labeled neurons. Dual whole-cell recordings reveal behavioral modulation of membrane potential synchrony and properties of synaptic transmission in vivo...
September 13, 2017: Neuron
https://www.readbyqxmd.com/read/28904098/characterization-of-ion-channels-and-o2-sensitivity-in-gill-neuroepithelial-cells-of-the-anoxia-tolerant-goldfish-carassius-auratus
#14
Peter C Zachar, Wen Pan, Michael G Jonz
The neuroepithelial cell (NEC) of the fish gill is an important model for O2 sensing in vertebrates; however, a complete picture of the chemosensory mechanisms in NECs is lacking, and O2 chemoreception in vertebrates that are tolerant to anoxia has yet not been explored. Using whole-cell patch-clamp recording, we characterized four types of ion channels in NECs isolated from the anoxia-tolerant goldfish. A Ca(2+)-dependent K(+) current (IKCa) peaked at ~20 mV, was potentiated by increased intracellular Ca(2+), and was reduced by 100 μM Cd(2+) A voltage-dependent inward current in Ba(2+) solution, with peak at 0 mV, confirmed the presence of Ca(2+) channels...
September 13, 2017: Journal of Neurophysiology
https://www.readbyqxmd.com/read/28902436/neuroprotective-effect-of-amantadine-on-corticosterone-induced-abnormal-glutamatergic-synaptic-transmission-of-ca3-ca1-pathway-in-rat-s-hippocampal-slices
#15
Xi Xiao, Hui Zhang, Hui Wang, Qun Li, Tao Zhang
Depression is a psychiatric disorder and chronic stress, leading to altered glucocorticoid secretion patterns, is one of the factors that induce depression. Our previous study showed that amantadine significantly attenuated the impairments of synaptic plasticity and cognitive function a rat model of CUS. However, little is known regarding the underlying mechanism. In the present study, the whole-cell patch-clamp technique was applied to examine the protection effect of amantadine on the hippocampus CA3-CA1 pathway...
September 13, 2017: Synapse
https://www.readbyqxmd.com/read/28900735/4-chloro-3-nitro-n-butylbenzenesulfonamide-acts-on-kv3-1-channels-by-an-open-channel-blocker-mechanism
#16
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...
September 12, 2017: Amino Acids
https://www.readbyqxmd.com/read/28898995/uncovering-the-arrhythmogenic-potential-of-trpm4-activation-in-atrial-derived-hl-1-cells-using-novel-recording-and-numerical-approaches
#17
Yaopeng Hu, Yubin Duan, Ayako Takeuchi, Lin Hai-Kurahara, Jun Ichikawa, Keizo Hiraishi, Tomohiro Numata, Hiroki Ohara, Gentaro Iribe, Michio Nakaya, Masayuki X Mori, Satoshi Matsuoka, Genshan Ma, Ryuji Inoue
Aims: Transient receptor potential cation channel subfamily melastatin member 4 (TRPM4), a Ca2+-activated nonselective cation channel abundantly expressed in the heart, has been implicated in conduction block and other arrhythmic propensities associated with cardiac remodelling and injury. The present study aimed to quantitatively evaluate the arrhythmogenic potential of TRPM4. Methods and results: Patch clamp and biochemical analyses were performed using expression system and an immortalized atrial cardiomyocyte cell line (HL-1), and numerical model simulation was employed...
August 1, 2017: Cardiovascular Research
https://www.readbyqxmd.com/read/28890050/modulation-of-hippocampal-synapse-maturation-by-activity-regulated-e3-ligase-via-non-canonical-pathway
#18
Pushpa Kumari, Balakumar Srinivasan, Sourav Banerjee
Development of functional synapses is crucial for the transmission and storage of information in the brain. Post establishment of the initial synaptic contact, synapses are stabilized through neuronal activity-induced signals. Emerging studies have implicated ubiquitination; a reversible posttranslational modification, as a key regulatory switch that modulates synapse development through proteasomal degradation. Ubiquitination of proteins is precisely regulated by E3 ligases, a set of enzymes that bind to specific substrates to facilitate the conjugation of monomeric or polymeric ubiquitin...
September 8, 2017: Neuroscience
https://www.readbyqxmd.com/read/28889099/the-impact-of-heterozygous-kcnk3-mutations-associated-with-pulmonary-arterial-hypertension-on-channel-function-and-pharmacological-recovery
#19
Michael S Bohnen, Danilo Roman-Campos, Cecile Terrenoire, Jack Jnani, Kevin J Sampson, Wendy K Chung, Robert S Kass
BACKGROUND: Heterozygous loss of function mutations in the KCNK3 gene cause hereditary pulmonary arterial hypertension (PAH). KCNK3 encodes an acid-sensitive potassium channel, which contributes to the resting potential of human pulmonary artery smooth muscle cells. KCNK3 is widely expressed in the body, and dimerizes with other KCNK3 subunits, or the closely related, acid-sensitive KCNK9 channel. METHODS AND RESULTS: We engineered homomeric and heterodimeric mutant and nonmutant KCNK3 channels associated with PAH...
September 9, 2017: Journal of the American Heart Association
https://www.readbyqxmd.com/read/28882594/terbinafine-is-a-novel-and-selective-activator-of-the-two-pore-domain-potassium-channel-task3
#20
Paul D Wright, Emma L Veale, David McCoull, David C Tickle, Jonathan M Large, Emma Ococks, Gemma Gothard, Catherine Kettleborough, Alistair Mathie, Jeffrey Jerman
Two-pore domain potassium channels (K2Ps) are characterized by their four transmembrane domain and two-pore topology. They carry background (or leak) potassium current in a variety of cell types. Despite a number of important roles there is currently a lack of pharmacological tools with which to further probe K2P function. We have developed a cell-based thallium flux assay, using baculovirus delivered TASK3 (TWIK-related acid-sensitive K(+) channel 3, KCNK9, K2P9.1) with the aim of identifying novel, selective TASK3 activators...
September 5, 2017: Biochemical and Biophysical Research Communications
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