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V L Zamoyski, V V Grigoriev
A decrease in the external chloride ion concentration to 4 mM caused a decrease in the fast sodium current to 85-100% in rat cerebellum Purkinje neurons in the whole-cell configuration using the patchclamp method. This effect did not depend on the main cation from the internal side of the cell membrane (120 mM of potassium or cesium) and also appeared when Cl- was substituted with sulfate (SO2 -4 ) or phosphate (H2 PO4 - ) anion outside of the cell. The effect was reversible after washing out the low chloride by the standard saline...
November 2017: Doklady. Biochemistry and Biophysics
Josef Buttigieg, Jie Pan, Herman Yeger, Ernest Cutz
Pulmonary neuroepithelial bodies (NEBs), composed of clusters of amine [serotonin (5-HT)] and peptide-producing cells, are widely distributed within the airway mucosa of human and animal lungs. NEBs are thought to function as airway O(2)-sensors, since they are extensively innervated and release 5-HT upon hypoxia exposure. The small cell lung carcinoma cell line (H146) provides a useful model for native NEBs, since they contain (and secrete) 5-HT and share the expression of a membrane-delimited O(2) sensor [classical NADPH oxidase (NOX2) coupled to an O(2)-sensitive K(+) channel]...
October 1, 2012: American Journal of Physiology. Lung Cellular and Molecular Physiology
Jun Ando, Nicholas I Smith, Katsumasa Fujita, Satoshi Kawata
We monitored femtosecond laser induced membrane potential changes in non-excitable cells using patchclamp analysis. Membrane potential hyperpolarization of HeLa cells was evoked by 780 nm, 80 fs laser pulses focused in the cellular cytoplasm at average powers of 30-60 mW. Simultaneous detection of intracellular Ca2+ concentration and membrane potential revealed coincident photogeneration of Ca2+ waves and membrane potential hyperpolarization. By using non-excitable cells, the cell dynamics are slow enough that we can calculate the membrane potential using the steady-state approximation for ion gradients and permeabilities, as formulated in the GHK equations...
February 2009: European Biophysics Journal: EBJ
F Homblé, J M Ferrier, J Dainty
Passive transport of potassium through the plasma membrane of a protoplasmic droplet isolated from large internodal cells of Chara corallina Klein ex Willd., em, R.D.W. has been investigated using the patchclamp technique. When the membrane is hyperpolarized the conductance of a single K(+)-channel is of the order of magnitude of 100 picoSiemens and is reduced by tetraethylammonium chloride. Its open time is voltage dependent. This voltage-dependent K(+)-channel displays rectifying properties. The channel density is about 0...
January 1987: Plant Physiology
Henrik Alle, Jörg R P Geiger
In the mammalian cortex, it is generally assumed that the output information of neurons is encoded in the number and the timing of action potentials. Here, we show, by using direct patchclamp recordings from presynaptic hippocampal mossy fiber boutons, that axons transmit analog signals in addition to action potentials. Excitatory presynaptic potentials result from subthreshold dendritic synaptic inputs, which propagate several hundreds of micrometers along the axon and modulate action potential-evoked transmitter release at the mossy fiber-CA3 synapse...
March 3, 2006: Science
William R Kem, Kristin Wildeboer, Susan LeFrancois, Ma'an Raja, William Marszalec, Jean-Claude Braekman
1. Tetraponerines are a group of alkaloids occurring in the venoms of ants belonging to the genus Tetraponera. Eight compounds had been isolated and their structures elucidated, but their mechanisms of action had not yet been reported. We have studied the actions of several of these tetraponerines on vertebrate neuromuscular, ganglionic, and brain nicotinic acetylcholine receptors (nAChRs) using a variety of techniques including muscle contracture, cultured cell functional assays, neuronal patch clamping, and radioligand binding methods...
August 2004: Cellular and Molecular Neurobiology
Jie Wu, Kevin Ellsworth, Marc Ellsworth, Katherine M Schroeder, Kris Smith, Robert S Fisher
Brain cholesterol synthesis inhibition (CSI) at a young age in rats has been shown to be a faithful model of acquired absence epilepsy, a devastating condition for which few therapies or models exist. We employed the CSI model to study cellular mechanisms of acquired absence epilepsy in Long-Evans Hooded rats. Patch-clamp, whole-cell recordings were compared from neurons acutely dissociated from the nucleus reticularis of thalamus (nRt) treated and untreated with a cholesterol synthesis inhibitor, U18666A. In U18666A-treated animals, 91% of rats developed EEG spike-waves (SWs)...
July 9, 2004: Brain Research
Denise Y Burton, Cunxian Song, Ilia Fishbein, Senator Hazelwood, Quanyi Li, Suzanne DeFelice, Jeanne M Connolly, Itay Perlstein, Douglas A Coulter, Robert J Levy
The present studies investigated the cardiac potassium channel missense mutation, Q9E-hMiRP1, for potential use as a gene therapy construct for cardiac arrhythmias. This gene abnormality is one of a number of mutations that can cause the long QT syndrome (LQTS), a hereditary arrhythmia disorder that is associated with sudden death. However, individuals who carry the Q9E-hMiRP1 variant are predisposed to developing the LQTS only after clarithromycin administration. Because the electrophysiologic mechanism of action of Q9E-hMiRP1 (i...
June 10, 2003: Human Gene Therapy
Q Lu, Z Wu, D Li
OBJECTIVE: To investigate whether apoptotic injury is induced by Fas mRNA expression mediated by H(2)O(2) and concerned with intracellular Ca(2+) unbalance at a single cell level. METHODS: Fas mRNA expression in human hepatocyte (LO(2) cell) was analyzed by whole-cell patch-clamp combined with single cell reverse transcription polymerase chain reaction (RT-PCR). Cytosolic Ca(2+) concentration was measured with another whole-cell patch-clamp combined with single-cell microfluorescence [Ca(2+)] i measurement at the same time...
March 2000: Zhonghua Yi Xue za Zhi [Chinese medical journal]
A Michalke, H J Galla, C Steinem
Solid-supported membranes immobilized on gold electrodes were used to detect and characterize the spontaneously inserting anion-selective protein channel (Clavibacter anion channel, CAC) present in the culture fluid of Clavibacter michiganense ssp. nebraskense. Three different membrane systems varying in the composition of the first chemisorbed monolayer were investigated by means of impedance spectroscopy. Conductance changes of the immobilized lipid membranes were sensitively detected after adding the culture fluid of the bacteria to the solid-supported membranes, indicating that the relative change in conductance is largest if the lipid layer is attached to the surface via a flexible lipid anchor...
October 2001: European Biophysics Journal: EBJ
W Qu, A J Moorhouse, A M Cunningham, P H Barry
Anomalous mole-fraction effects (AMFE) were studied, using the inside-out configuration of the patchclamp technique, in both recombinant wild-type alpha-homomeric rat olfactory adenosine 3',5'-cyclic monophosphate (cAMP)-gated channels (rOCNC1) expressed in human embryonic kidney cells (HEK 293) and native cyclic nucleotide-gated (CNG) channels in acutely isolated rat olfactory receptor neurons. Single-channel and macroscopic currents were activated by 200 microM and 500 microM cAMP, respectively. Macroscopic currents, measured with mixtures of Na(+)-NH(4)(+) or Cs(+)-Li(+) in the cytoplasmic bathing solution, displayed AMFE in the rOCNC1 channels at both positive and negative membrane potentials...
July 7, 2001: Proceedings. Biological Sciences
J Zheng, W N Zagotta
Site-specific fluorescence recordings have shown great promise in understanding conformational changes in signaling proteins. The reported applications on ion channels have been limited to extracellular sites in whole oocyte preparations. We are now able to directly monitor gating movements of the intracellular domains of cyclic nucleotide-gated channels using simultaneous site-specific fluorescence recording and patchclamp current recording from inside-out patches. Fluorescence signals were reliably observed when fluorophore was covalently attached to a site between the cyclic nucleotide-binding domain and the pore...
November 2000: Neuron
J Lee, W Lim, S Y Eun, S J Kim, J Kim
The effect of hypoxia on small-conductance Ca(2+)-activated K+ current was investigated in a study of adult rat adrenomedullary chromaffin cells (AMCs), which were maintained in short-term culture. The nystatin-perforated, whole-cell patchclamp technique was used to study the effect of hypoxia with minimum perturbation of the intracellular milieu. Under voltage-clamp conditions, acute hypoxia (P(O2) approximately equal to 25 mmHg) suppressed the whole-cell outward currents of more than half the AMCs (24/46)...
April 2000: Pflügers Archiv: European Journal of Physiology
Y A Mei, L X Huang, H Wei, J T Sun, H Q Zhou, Z H Zhang
AIM: To characterize a voltage-dependent outward K+ current in cultured heart cells of 14-16-day-old embryos of yellow chick. METHODS: The patchclamp technique in the whole-cell configuration was used. RESULTS: The kinetics and the pharmacology of the outward K+ current in our cell mold were different from those described in white chick. Like the calcium-activated K+ current, blocker of calcium channel, CdCl2, eliminated the current of more than 95%...
April 1999: Zhongguo Yao Li Xue Bao, Acta Pharmacologica Sinica
R Skryma, F Van Coppenolle, L Dufy-Barbe, B Dufy, N Prevarskaya
Potassium plasma membrane channels have been studied in the LNCaP androgen-sensitive human prostate cancer cell line, derived from a lymph node of a subject with metastatic carcinoma of the prostate. Membrane currents were recorded by the patchclamp technique, using the cell-attached, cell-free and whole-cell mode. A voltage-dependent, non-inactivating potassium channel (delayed rectifier) was the most commonly observed ion channel in LNCaP cells. The slope conductance of K+ channels in a symmetrical 140 mM K+ gradient was 78 pS...
1999: Receptors & Channels
M Heine, D Wicher
The effect of a Ca2+ resting current on spiking and intracellular Ca2+ concentration [Ca2+]i of isolated cockroach dorsal unpaired median neurons was investigated by combining microfluometry (Fura2) with patchclamping. The resting current was reduced by 10 microM 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) and 10 microM mefenamic acid, which caused slower spiking of cells. Up-modulation of the resting current by neurohormone D (NHD, 100 fM to 10 pM) transiently increased [Ca2+]i. This Ca2+ transient was largely reduced by the ryanodine receptor antagonist dantrolene (10 microM) but it was not reduced by the IP3-receptor antagonist heparin which indicates Ca2+-induced Ca2+ release...
October 5, 1998: Neuroreport
J Krupp, A Bordey, P Feltz
The time pattern of glycinergic inhibitory postsynaptic currents (IPSCs) in sympathetic preganglionic neurons was studied in thin transverse spinal cord slices of neonatal (1-10 days postnatal) rats by means of the patchclamp technique. Three time patterns could be distinguished: (i) large events [mostly > 400 pA (30-36 degrees C)] occurring at regular intervals, (ii) small events occurring at irregular intervals, and (iii) small events occurring in transient (1.5-10 s), high-frequency (> 15 Hz) bursts of synaptic activity...
August 1997: European Journal of Neuroscience
K Imon, T Amano, K Ishihara, M Sasa, K Yajin
To determine whether functional Ca2+ channels are present in vestibular dark cells, changes in intracellular Ca2+ concentration ([Ca2+]i) due to K+ applications were measured using the Ca(2+)-sensitive dye (fura-2) and patchclamp whole-cell recordings were made in dark cells isolated from the ampullae of the semicircular canal of the guinea pig. Exchange of the external solution with a buffer medium containing a high K+ concentration (80 mM K+ or 150 mM K+) caused a concentration-dependent increase in [Ca2+]i in vestibular dark cells...
1997: European Archives of Oto-rhino-laryngology
C U Cotton, L al-Nakkash
We describe a method to isolate and culture epithelial cells from the main duct of the bovine pancreas. In primary cultures, secretin caused a dose-dependent increase in intracellular adenosine 3',5'-cyclic monophosphate (cAMP) and stimulated electrogenic transepithelial ion transport. Elevation of intracellular cAMP increased the rate coefficient for 36Cl- efflux from 0.14 +/- 0.03 to 0.47 +/- 0.12 min-1, and plasma membrane conductance, measured by the whole cell patchclamp technique, was increased from 0...
June 1997: American Journal of Physiology
C Cokmus, E W Davidson, K Cooper
The electrophysiological effects of both trypsinactivated native and 42- and 51-kDa cloned binary toxins of Bacillus sphaericus were investigated on cultured Culex quinquefasciatus cells using the patchclamp technique. Rates of reduction in whole-cell membrane resistance were correlated with increasing native toxin concentration. The 42- or 51-kDa cloned toxin alone at 50 micrograms/ml reduced the resistance. Electrophysiological effects occurred before any changes were visible by phase-contrast microscopy.
May 1997: Journal of Invertebrate Pathology
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