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Patch clamp nAChR

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https://www.readbyqxmd.com/read/27856558/human-polymorphisms-in-nicotinic-receptors-a-functional-analysis-in-ips-derived-dopaminergic-neurons
#1
Cristina Deflorio, Stéphane Blanchard, Maria Carla Carisì, Delphine Bohl, Uwe Maskos
Tobacco smoking is a public health problem, with ∼5 million deaths per year, representing a heavy burden for many countries. No effective therapeutic strategies are currently available for nicotine addiction, and it is therefore crucial to understand the etiological and pathophysiological factors contributing to this addiction. The neuronal α5 nicotinic acetylcholine receptor (nAChR) subunit is critically involved in nicotine dependence. In particular, the human polymorphism α5D398N corresponds to the strongest correlation with nicotine dependence risk found to date in occidental populations, according to meta-analysis of genome-wide association studies...
November 17, 2016: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/27805736/therapeutic-concentrations-of-varenicline-in-the-presence-of-nicotine-increase-action-potential-firing-in-human-adrenal-chromaffin-cells
#2
Arik J Hone, J Michael McIntosh, Lola Rueda-Ruzafa, Juan Passas, Cristina de Castro-Guerín, Jesús Blázquez, Carmen González-Enguita, Almudena Albillos
Varenicline is a nicotinic acetylcholine receptor (nAChR) agonist used to treat nicotine addiction but a live debate persists concerning its mechanism of action in reducing nicotine consumption. Although initially reported as α4β2-selective, varenicline was subsequently shown to activate other nAChR subtypes implicated in nicotine addiction including α3β4. However, it remains unclear whether activation of α3β4 nAChRs by therapeutically relevant concentrations of varenicline is sufficient to affect the behavior of cells that express this subtype...
November 2, 2016: Journal of Neurochemistry
https://www.readbyqxmd.com/read/27505073/electrophysiology-based-assays-to-detect-subtype-selective-modulation-of-human-nicotinic-acetylcholine-receptors
#3
Glenn E Kirsch, Nikolai B Fedorov, Yuri A Kuryshev, Zhiqi Liu, Lucas C Armstrong, Michael S Orr
The Family Smoking Prevention and Tobacco Control Act of 2009 (Public Law 111-31) gave the US Food and Drug Administration (FDA) the responsibility for regulating tobacco products. Nicotine is the primary addictive component of tobacco and its effects can be modulated by additional ingredients in manufactured products. Nicotine acts by mimicking the neurotransmitter acetylcholine on neuronal nicotinic acetylcholine receptors (nAChRs), which function as ion channels in cholinergic modulation of neurotransmission...
August 2016: Assay and Drug Development Technologies
https://www.readbyqxmd.com/read/27477858/influence-of-developmental-nicotine-exposure-on-glutamatergic-neurotransmission-in-rhythmically-active-hypoglossal-motoneurons
#4
Marina Cholanian, Gregory L Powell, Richard B Levine, Ralph F Fregosi
Developmental nicotine exposure (DNE) is associated with increased risk of cardiorespiratory, intellectual, and behavioral abnormalities in neonates, and is a risk factor for apnea of prematurity, altered arousal responses and Sudden Infant Death Syndrome. Alterations in nicotinic acetylcholine receptor signaling (nAChRs) after DNE lead to changes in excitatory neurotransmission in neural networks that control breathing, including a heightened excitatory response to AMPA microinjection into the hypoglossal motor nucleus...
July 29, 2016: Experimental Neurology
https://www.readbyqxmd.com/read/27374167/nicotinic-receptor-activation-contrasts-pathophysiological-bursting-and-neurodegeneration-evoked-by-glutamate-uptake-block-on-rat-hypoglossal-motoneurons
#5
Silvia Corsini, Maria Tortora, Andrea Nistri
KEY POINTS: Impaired uptake of glutamate builds up the extracellular level of this excitatory transmitter to trigger rhythmic neuronal bursting and delayed cell death in the brainstem motor nucleus hypoglossus. This process is the expression of the excitotoxicity that underlies motoneuron degeneration in diseases such as amyotrophic lateral sclerosis affecting bulbar motoneurons. In a model of motoneuron excitotoxicity produced by pharmacological block of glutamate uptake in vitro, rhythmic bursting is suppressed by activation of neuronal nicotinic receptors with their conventional agonist nicotine...
November 15, 2016: Journal of Physiology
https://www.readbyqxmd.com/read/27336596/mutation-linked-to-autosomal-dominant-nocturnal-frontal-lobe-epilepsy-reduces-low-sensitivity-%C3%AE-4%C3%AE-2-and-increases-%C3%AE-5%C3%AE-4%C3%AE-2-nicotinic-receptor-surface-expression
#6
Weston A Nichols, Brandon J Henderson, Christopher B Marotta, Caroline Y Yu, Chris Richards, Dennis A Dougherty, Henry A Lester, Bruce N Cohen
A number of mutations in α4β2-containing (α4β2*) nicotinic acetylcholine (ACh) receptors (nAChRs) are linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE), including one in the β2 subunit called β2V287L. Two α4β2* subtypes with different subunit stoichiometries and ACh sensitivities co-exist in the brain, a high-sensitivity subtype with (α4)2(β2)3 subunit stoichiometry and a low-sensitivity subtype with (α4)3(β2)2 stoichiometry. The α5 nicotinic subunit also co-assembles with α4β2 to form a high-sensitivity α5α4β2 nAChR...
2016: PloS One
https://www.readbyqxmd.com/read/27260673/inhibition-by-ketamine-and-amphetamine-analogs-of-the-neurogenic-nitrergic-vasodilations-in-porcine-basilar-arteries
#7
Mei-Fang Chen, Su-Yu Lai, Po-Cheng Kung, Yo-Cheng Lin, Hui-I Yang, Po-Yi Chen, Ingrid Y Liu, Ahai Chang Lua, Tony Jer-Fu Lee
The abuse of ketamine and amphetamine analogs is associated with incidence of hypertension and strokes involving activation of sympathetic activities. Large cerebral arteries at the base of the brain from several species receive dense sympathetic innervation which upon activation causes parasympathetic-nitrergic vasodilation with increased regional blood flow via axo-axonal interaction mechanism, serving as a protective mechanism to meet O2 demand in an acutely stressful situation. The present study was designed to examine effects of ketamine and amphetamine analogs on axo-axonal interaction-mediated neurogenic nitrergic vasodilation in porcine basilar arteries using techniques of blood-vessel myography, patch clamp and two-electrode voltage clamp, and calcium imaging...
August 15, 2016: Toxicology and Applied Pharmacology
https://www.readbyqxmd.com/read/27256075/nicotine-inhibits-activation-of-microglial-proton-currents-via-interactions-with-%C3%AE-7-acetylcholine-receptors
#8
Mami Noda, A I Kobayashi
Alpha 7 subunits of nicotinic acetylcholine receptors (nAChRs) are expressed in microglia and are involved in the suppression of neuroinflammation. Over the past decade, many reports show beneficial effects of nicotine, though little is known about the mechanism. Here we show that nicotine inhibits lipopolysaccharide (LPS)-induced proton (H(+)) currents and morphological change by using primary cultured microglia. The H(+) channel currents were measured by whole-cell patch clamp method under voltage-clamp condition...
June 2, 2016: Journal of Physiological Sciences: JPS
https://www.readbyqxmd.com/read/27233449/analysis-of-neuronal-nicotinic-acetylcholine-receptor-%C3%AE-4%C3%AE-2-activation-at-the-single-channel-level
#9
Camila Carignano, Esteban Pablo Barila, Guillermo Spitzmaul
The neuronal nicotinic acetylcholine receptor α4β2 forms pentameric proteins with two alternate stoichiometries. The high-sensitivity receptor is related to (α4)2(β2)3 stoichiometry while the low-sensitivity receptor to (α4)3(β2)2 stoichiometry. Both subtypes share two binding sites at the α4((+))/β2((-)) interface with high affinity for agonists. (α4)3(β2)2 has an additional binding site at the α4((+))/α4((-)) interface with low affinity for agonists. We investigated activation kinetics of both receptor subtypes by patch-clamp recordings of single-channel activity in the presence of several concentrations of acetylcholine (0...
September 2016: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/26806304/differential-modulation-of-gabaa-and-nmda-receptors-by-%C3%AE-7-nicotinic-receptor-desensitization-in-cultured-rat-hippocampal-neurons
#10
Lei Shen, Wen-yu Cui, Ru-zhu Chen, Hai Wang
AIM: To explore the modulatory effect of desensitized α7-containing nicotinic receptors (α7-nAChRs) on excitatory and inhibitory amino acid receptors in cultured hippocampal neurons and to identify the mechanism underlying this effect. METHODS: Whole-cell patch-clamp recordings were performed on cultured rat hippocampal neurons to measure α7-nAChR currents and to determine the role of desensitized α7-nAChRs on brain amino acid receptor activity. RESULTS: Pulse and perfusion applications of the α7-nAChR agonist choline were applied to induce different types of α7-nAChR desensitization in cultured hippocampal neurons...
March 2016: Acta Pharmacologica Sinica
https://www.readbyqxmd.com/read/26585318/nicotinic-acetylcholine-receptors-nachrs-at-zebrafish-red-and-white-muscle-show-different-properties-during-development
#11
Kazi T Ahmed, Declan W Ali
Nicotinic acetylcholine receptors (nAChRs) are highly expressed at the vertebrate neuromuscular junction (NMJ) where they are required for muscle activation. Understanding the factors that underlie NMJ development is critical for a full understanding of muscle function. In this study we performed whole cell and outside-out patch clamp recordings, and single-cell RT-qPCR from zebrafish red and white muscle to examine the properties of nAChRs during the first 5 days of development. In red fibers miniature endplate currents (mEPCs) exhibit single exponential time courses at 1...
August 2016: Developmental Neurobiology
https://www.readbyqxmd.com/read/26441658/agonist-and-antagonist-effects-of-tobacco-related-nitrosamines-on-human-%C3%AE-4%C3%AE-2-nicotinic-acetylcholine-receptors
#12
Simone Brusco, Paola Ambrosi, Simone Meneghini, Andrea Becchetti
Regulation of the "neuronal" nicotinic acetylcholine receptors (nAChRs) is implicated in both tobacco addiction and smoking-dependent tumor promotion. Some of these effects are caused by the tobacco-derived N-nitrosamines, which are carcinogenic compounds that avidly bind to nAChRs. However, the functional effects of these drugs on specific nAChR subtypes are largely unknown. By using patch-clamp methods, we tested 4-(methylnitrosamine)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN) on human α4β2 nAChRs...
2015: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/26354438/lateral-diffusion-function-and-expression-of-the-slow-channel-congenital-myasthenia-syndrome-%C3%AE-c418w-nicotinic-receptor-mutation-with-changes-in-lipid-raft-components
#13
Jessica Oyola-Cintrón, Daniel Caballero-Rivera, Leomar Ballester, Carlos A Baéz-Pagán, Hernán L Martínez, Karla P Vélez-Arroyo, Orestes Quesada, José A Lasalde-Dominicci
Lipid rafts, specialized membrane microdomains in the plasma membrane rich in cholesterol and sphingolipids, are hot spots for a number of important cellular processes. The novel nicotinic acetylcholine receptor (nAChR) mutation αC418W, the first lipid-exposed mutation identified in a patient that causes slow channel congenital myasthenia syndrome was shown to be cholesterol-sensitive and to accumulate in microdomains rich in the membrane raft marker protein caveolin-1. The objective of this study is to gain insight into the mechanism by which lateral segregation into specialized raft membrane microdomains regulates the activable pool of nAChRs...
October 30, 2015: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/26330550/%C3%AE-conotoxins-identify-the-%C3%AE-3%C3%AE-4-subtype-as-the-predominant-nicotinic-acetylcholine-receptor-expressed-in-human-adrenal-chromaffin-cells
#14
Arik J Hone, J Michael McIntosh, Layla Azam, Jon Lindstrom, Linda Lucero, Paul Whiteaker, Juan Passas, Jesús Blázquez, Almudena Albillos
Ligands that selectively inhibit human α3β2 and α6β2 nicotinic acetylcholine receptor (nAChRs) and not the closely related α3β4 and α6β4 subtypes are lacking. Current α-conotoxins (α-Ctxs) that discriminate among these nAChR subtypes in rat fail to discriminate among the human receptor homologs. In this study, we describe the development of α-Ctx LvIA(N9R,V10A) that is 3000-fold more potent on oocyte-expressed human α3β2 than α3β4 and 165-fold more potent on human α6/α3β2β3 than α6/α3β4 nAChRs...
November 2015: Molecular Pharmacology
https://www.readbyqxmd.com/read/26283025/inhibitory-actions-of-bisabolol-on-%C3%AE-7-nicotinic-acetylcholine-receptors
#15
S Nurulain, T Prytkova, A M Sultan, O Ievglevskyi, D Lorke, K-H S Yang, G Petroianu, F C Howarth, N Kabbani, M Oz
Bisabolol is a plant-derived monocyclic sesquiterpene alcohol with antinociceptive and antiinflammatory actions. However, molecular targets mediating these effects of bisabolol are poorly understood. In this study, using a two-electrode voltage-clamp and patch-clamp techniques and live cellular calcium imaging, we have investigated the effect of bisabolol on the function of human α7 subunit of nicotinic acetylcholine receptor (nAChR) in Xenopus oocytes, interneurons of rat hippocampal slices. We have found that bisabolol reversibly and concentration dependently (IC50 = 3...
October 15, 2015: Neuroscience
https://www.readbyqxmd.com/read/26210579/%C3%AE-6-containing-nicotinic-acetylcholine-receptors-in-midbrain-dopamine-neurons-are-poised-to-govern-dopamine-mediated-behaviors-and-synaptic-plasticity
#16
J N Berry, S E Engle, J M McIntosh, R M Drenan
Acetylcholine (ACh) acts through nicotinic and muscarinic ACh receptors in the ventral midbrain and striatal areas to influence dopamine (DA) transmission. This cholinergic control of DA transmission is important for processes such as attention and motivated behavior, and is manipulated by nicotine in tobacco products. Identifying and characterizing the key ACh receptors involved in cholinergic control of DA transmission could lead to small molecule therapeutics for treating disorders involving attention, addiction, Parkinson's disease, and schizophrenia...
September 24, 2015: Neuroscience
https://www.readbyqxmd.com/read/26162864/at-1001-is-a-partial-agonist-with-high-affinity-and-selectivity-at-human-and-rat-%C3%AE-3%C3%AE-4-nicotinic-cholinergic-receptors
#17
Edward W Tuan, Andrew G Horti, Thao T Olson, Yongiun Gao, Craig A Stockmeier, Nour Al-Muhtasib, Carrie Bowman Dalley, Amanda E Lewin, Barry B Wolfe, Niaz Sahibzada, Yingxian Xiao, Kenneth J Kellar
AT-1001 [N-(2-bromophenyl)-9-methyl-9-azabicyclo[3.3.1] nonan-3-amine] is a high-affinity and highly selective ligand at α3β4 nicotinic cholinergic receptors (nAChRs) that was reported to decrease nicotine self-administration in rats. It was initially reported to be an antagonist at rat α3β4 nAChRs heterologously expressed in HEK293 cells. Here we compared AT-1001 actions at rat and human α3β4 and α4β2 nAChRs similarly expressed in HEK 293 cells. We found that, as originally reported, AT-1001 is highly selective for α3β4 receptors over α4β2 receptors, but its binding selectivity is much greater at human than at rat receptors, because of a higher affinity at human than at rat α3β4 nAChRs...
October 2015: Molecular Pharmacology
https://www.readbyqxmd.com/read/26039516/nicotinic-acetylcholine-receptor-nachr-dependent-chorda-tympani-taste-nerve-responses-to-nicotine-ethanol-and-acetylcholine
#18
Zuo Jun Ren, Shobha Mummalaneni, Jie Qian, Clive M Baumgarten, John A DeSimone, Vijay Lyall
Nicotine elicits bitter taste by activating TRPM5-dependent and TRPM5-independent but neuronal nAChR-dependent pathways. The nAChRs represent common targets at which acetylcholine, nicotine and ethanol functionally interact in the central nervous system. Here, we investigated if the nAChRs also represent a common pathway through which the bitter taste of nicotine, ethanol and acetylcholine is transduced. To this end, chorda tympani (CT) taste nerve responses were monitored in rats, wild-type mice and TRPM5 knockout (KO) mice following lingual stimulation with nicotine free base, ethanol, and acetylcholine, in the absence and presence of nAChR agonists and antagonists...
2015: PloS One
https://www.readbyqxmd.com/read/26038197/cholinergic-enhancement-of-cell-proliferation-in-the-postnatal-neurogenic-niche-of-the-mammalian-spinal-cord
#19
Laura F Corns, Lucy Atkinson, Jill Daniel, Ian J Edwards, Lauryn New, Jim Deuchars, Susan A Deuchars
The region surrounding the central canal (CC) of the spinal cord is a highly plastic area, defined as a postnatal neurogenic niche. Within this region are ependymal cells that can proliferate and differentiate to form new astrocytes and oligodendrocytes following injury and cerebrospinal fluid contacting cells (CSFcCs). The specific environmental conditions, including the modulation by neurotransmitters that influence these cells and their ability to proliferate, are unknown. Here, we show that acetylcholine promotes the proliferation of ependymal cells in mice under both in vitro and in vivo conditions...
September 2015: Stem Cells
https://www.readbyqxmd.com/read/25959067/a-novel-nicotinic-mechanism-underlies-%C3%AE-amyloid-induced-neurotoxicity
#20
Qiang Liu, Xitao Xie, Sharareh Emadi, Michael R Sierks, Jie Wu
Loss of basal forebrain cholinergic neurons (BFCN) correlates with cognitive deficits in Alzheimer disease (AD). Our recent evidence suggests that chronic exposure to Aβ up-regulated neuronal α7-nAChRs and increased neuronal excitability in cultured hippocampal neurons. However, the impact of the up-regulated α7-nAChRs on Aβ-induced neurotoxicity remains unclear. In this study, we investigated the role of α7-nAChRs in the mediation of Aβ-induced neurotoxicity. The effects of Aβ exposure on α7-nAChRs and cytotoxicity were examined using whole-cell patch clamp recordings, atomic force microscope (AFM) imaging, immunoprecipitation, and lactate dehydrogenase (LDH) release assay in primary cultured hippocampal neurons as well as differentiated human neuroblastoma (SH-SY5Y) cells with cholinergic characteristics...
October 2015: Neuropharmacology
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