cAMP-activated chloride conductance in the colonic cell line, Caco-2

1992 ◽  
Vol 262 (1) ◽  
pp. C251-C256 ◽  
Author(s):  
C. E. Bear ◽  
E. F. Reyes
Keyword(s):  
Reversal Potential ◽  
Channel Activity ◽  
Cell Potential ◽  
Linear Current ◽  
Current Voltage ◽  
Patch Pipette ◽  
Cl Channels ◽  
Patch Clamp Electrophysiology ◽  
Best Fit ◽  
Linear Regressions

In this study we investigated the properties of adenosine 3',5'-cyclic monophosphate (cAMP)-stimulated Cl- efflux in Caco-2 monolayers by measuring 125I efflux rates from preloaded cells and using patch-clamp electrophysiology. The addition of a cocktail containing 100 microM dibutyryl cAMP (DBcAMP), 10 microM forskolin, and 1 mM 3-isobutyl-1-methylxanthine caused a significant (P less than 0.05) increase in the rate of 125I efflux. Dissipation of cell potential by adding valinomycin (4.5 microM) with 135 mM extracellular KCl reduced the cAMP-evoked 125I efflux. These results suggest that cAMP-stimulated anion efflux occurs through a conductive pore or channel. Whole cell currents evoked with DBcAMP or forskolin were anion selective, PCl greater than PI greater than Pgluconate, and exhibited a linear current-voltage (I-V) relationship. Currents evoked with depolarizing or hyperpolarizing voltage steps showed no evidence of time-dependent activation or inactivation. Single Cl- channels were stimulated in cell-attached patches after treatment with cAMP. Onset of channel activity occurred after 20-30s of cAMP treatment, and the response was long lasting. The I-V relationship for the channel activated in cell-attached patches by cAMP was best fit using two linear regressions. The slope conductance of the channel was 3.2 +/- 0.6 and 7.4 +/- 0.3 pS at hyperpolarizing and depolarizing potentials, respectively. Substitution of 140 mM NaCl with 70 mM NaCl in the patch pipette resulted in a positive shift in reversal potential, indicating that the channel is anion selective.(ABSTRACT TRUNCATED AT 250 WORDS)

Ca(2+)-activated nonselective cation channel in apical membrane of vestibular dark cells

1992 ◽  
Vol 262 (6) ◽  
pp. C1423-C1429 ◽  
Author(s):  
D. C. Marcus ◽  
S. Takeuchi ◽  
P. Wangemann
Keyword(s):  
Single Channel ◽  
Apical Membrane ◽  
Cation Channel ◽  
Nonselective Cation Channel ◽  
Channel Activity ◽  
Dark Cells ◽  
Linear Current ◽  
Current Voltage ◽  
K Secretion ◽  
Vestibular Dark Cells

Patch-clamp recordings were made on cell-attached and excised apical membrane from dark cells of the semicircular canal of the gerbil. These cells are thought to secrete K+ and absorb Na+ from the luminal fluid (endolymph). Single-channel events were identified as being equally conductive (27.6 +/- 0.4 pS; n = 48) for K+, Na+, Rb+, Li+, and Cs+ and 1.4 times more permeable to NH4+ but not permeable to Cl-, Ca2+, Ba2+, nor to N-methyl-D-glucamine. The channels displayed linear current-voltage relations that passed nearly through the origin (intercept: -2.6 +/- 0.5 mV; n = 48) when conductive monovalent cations were present on both sides of the membrane in equal concentrations. Channel activity required the presence of Ca2+ at the cytosolic face; there was no activity at less than or equal to 10(-7) M Ca2+ and full activity at greater than or equal to 10(-5) M Ca2+. Cell-attached recordings had a mean reversal voltage of -36.4 +/- 7.9 mV (n = 7), which was interpreted to reflect the intracellular potential of dark cells under the present conditions. We have identified a nonselective cation channel in the apical membrane of vestibular dark cells that might participate in K+ secretion or Na+ absorption under stimulated conditions, but the density appears to be insufficient to fully account for the transepithelial K+ flux.

NMDA-receptor-mediated synaptic currents in guinea pig laterodorsal tegmental neurons in vitro

1996 ◽  
Vol 76 (2) ◽  
pp. 1101-1111 ◽  
Author(s):  
R. Sanchez ◽  
C. S. Leonard
Keyword(s):  
Nmda Receptor ◽  
Guinea Pig ◽  
Nmda Receptors ◽  
Brain Stem ◽  
Reversal Potential ◽  
Negative Slope ◽  
Synaptic Currents ◽  
Linear Current ◽  
Current Voltage ◽  
Current Voltage Relation

1. Whole cell voltage-clamp techniques were used to record glutamate-receptor-mediated synaptic currents from neurons of the laterodorsal tegmental nucleus (LDT). The principal cells of the LDT contain acetylcholine and nitric oxide synthase, and are believed to be involved in the control of sleep-waking behavior via widespread projections to the thalamus and brain stem. LDT cells were recorded from slices of mature guinea pig brain stem with patch pipette solutions containing cesium as the primary cation. 2. Application of N-methyl-D-aspartate (NMDA) elicited currents that were strongly voltage dependent with a mean reversal potential of +16.3 mV. Peak currents occurred near -15 mV, and a region of negative slope conductance was seen at more negative potentials. Application of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid evoked currents that exhibited a nearly linear current-voltage relation with a mean reversal potential of -3.4 mV. 3. Electrical stimulation of local afferents elicited dual-component excitatory postsynaptic currents (EPSCs) with decays that were well fitted by the sum of two exponentials. Mean decay time constants at -60 mV were 8.77 ms for the faster component and 129.4 ms for the slower component. The faster component displayed a linear current-voltage relation and was blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) or 6,7-dinitroquinoxaline-2,3-dione, indicating that it was mediated by non-NMDA receptors, whereas the slower component displayed a voltage dependence similar to that for NMDA-evoked currents and was blocked by 2-amino-5-phosphonopentanoic acid (AP-5), indicating its mediation by NMDA receptors. 4. The fractional contribution of NMDA receptors to the EPSC was estimated from double-exponential curve fits to the decay phases. With this method, NMDA receptors were estimated on average to carry 10.1% of the total peak EPSC at -60 mV. Blockade of the non-NMDA-receptor-mediated component with CNQX revealed a residual EPSC whose amplitude was 14.4% of the control value, whereas AP-5 alone reduced the control EPSC peak by 16.1%, both values were comparable with those obtained from curve fit estimates. 5. Previous work has shown that the presence of 4-aminopyridine-sensitive, A-like transient current in LDT cells is correlated with the cholinergic phenotype. The majority of cells in this study exhibited A-like transient currents that were blocked by 4-amino-pyridine, suggesting that the majority of the data were obtained from the cholinergic and NOS-containing neurons of the LDT nucleus. 6. These experiments demonstrate the synaptic activation of functional NMDA and non-NMDA receptors in LDT neurons, and indicate that NMDA receptors contribute to fast excitatory transmission in these cells. The results suggest that afferents releasing excitatory amino acids may play an important role in controlling the state-dependent activity of LDT neurons.

More than One Type of Conductance is Activated During Responses of Blowfly Monopolar Neurones

1989 ◽  
Vol 144 (1) ◽  
pp. 147-154 ◽  
Author(s):  
M. WECKSTRÖM ◽  
E. KOUVALAINEN ◽  
K. DJUPSUND ◽  
M. JÄRVILEHTO
Keyword(s):  
Compound Eye ◽  
Reversal Potential ◽  
Input Resistance ◽  
Current Clamp ◽  
Voltage Range ◽  
Continuous Release ◽  
Linear Current ◽  
Current Voltage ◽  
Current Voltage Relationship ◽  
Voltage Relationship

The principal second-order neurones in the blowfly compound eye, the large monopolar neurones (LMCs), were studied using intracellular recording and discontinuous current-clamp techniques, in combination with measurement of dynamic input resistance. The LMCs had resting potentials of −35 to −45 mV and showed a linear current-voltage relationship in the lamina in the physiological voltage range. The hyperpolarizing light-on transient was associated with a drop in input resistance from 17 ± 5 to 3 ± 1MΩ, and had a reversal potential between −60 and −90 mV. The dynamic input resistance of saturated responses and the properties of reversed responses suggested that more than one conductance was activated during the response of the LMCs. In lamina recordings, the input resistance increased beyond the resting level during repolarization, which can be interpreted in terms of a continuous release of transmitter by the photoreceptor terminals, even in darkness. The input resistance of LMCs in axon recordings in darkness and during the light-on response was generally higher than in the lamina recordings. The responses to light in axons also differed from those recorded in lamina by showing regenerative properties.

scholarly journals Lack of negative slope in I-V plots for BK channels at positive potentials in the absence of intracellular blockers

2013 ◽  
Vol 141 (4) ◽  
pp. 493-497 ◽  
Author(s):  
Yanyan Geng ◽  
Xiaoyu Wang ◽  
Karl L. Magleby
Keyword(s):  
Single Channel ◽  
Bk Channels ◽  
Negative Slope ◽  
Channel Conductance ◽  
Single Channel Conductance ◽  
Linear Current ◽  
Joint Application ◽  
Current Voltage ◽  
Α Subunits

Large-conductance, voltage- and Ca2+-activated K+ (BK) channels display near linear current–voltage (I-V) plots for voltages between −100 and +100 mV, with an increasing sublinearity for more positive potentials. As is the case for many types of channels, BK channels are blocked at positive potentials by intracellular Ca2+ and Mg2+. This fast block progressively reduces single-channel conductance with increasing voltage, giving rise to a negative slope in the I-V plots beyond about +120 mV, depending on the concentration of the blockers. In contrast to these observations of pronounced differences in the magnitudes and shapes of I-V plots in the absence and presence of intracellular blockers, Schroeder and Hansen (2007. J. Gen. Physiol. http://dx.doi.org/10.1085/jgp.200709802) have reported identical I-V plots in the absence and presence of blockers for BK channels, with both plots having reduced conductance and negative slopes, as expected for blockers. Schroeder and Hansen included both Ca2+ and Mg2+ in the intracellular solution rather than a single blocker, and they also studied BK channels expressed from α plus β1 subunits, whereas most previous studies used only α subunits. Although it seems unlikely that these experimental differences would account for the differences in findings between previous studies and those of Schroeder and Hansen, we repeated the experiments using BK channels comprised of α plus β1 subunits with joint application of 2.5 mM Ca2+ plus 2.5 mM Mg2+, as Schroeder and Hansen did. In contrast to the findings of Schroeder and Hansen of identical I-V plots, we found marked differences in the single-channel I-V plots in the absence and presence of blockers. Consistent with previous studies, we found near linear I-V plots in the absence of blockers and greatly reduced currents and negative slopes in the presence of blockers. Hence, studies of conductance mechanisms for BK channels should exclude intracellular Ca2+/Mg2+, as they can reduce conductance and induce negative slopes.

scholarly journals Plasticity of calcium-permeable AMPA glutamate receptors in Pro-opiomelanocortin neurons

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Shigetomo Suyama ◽  
Alexandra Ralevski ◽  
Zhong-Wu Liu ◽  
Marcelo O Dietrich ◽  
Toshihiko Yada ◽  
...  
Keyword(s):  
Food Deprivation ◽  
High Fat Diet ◽  
Receptor Complex ◽  
High Fat ◽  
Fasted State ◽  
Linear Current ◽  
Current Voltage ◽  
Relationship Of ◽  
Current Voltage Relationship ◽  
Voltage Relationship

POMC neurons integrate metabolic signals from the periphery. Here, we show in mice that food deprivation induces a linear current-voltage relationship of AMPAR-mediated excitatory postsynaptic currents (EPSCs) in POMC neurons. Inhibition of EPSCs by IEM-1460, an antagonist of calcium-permeable (Cp) AMPARs, diminished EPSC amplitude in the fed but not in the fasted state, suggesting entry of GluR2 subunits into the AMPA receptor complex during food deprivation. Accordingly, removal of extracellular calcium from ACSF decreased the amplitude of mEPSCs in the fed but not the fasted state. Ten days of high-fat diet exposure, which was accompanied by elevated leptin levels and increased POMC neuronal activity, resulted in increased expression of Cp-AMPARs on POMC neurons. Altogether, our results show that entry of calcium via Cp-AMPARs is inherent to activation of POMC neurons, which may underlie a vulnerability of these neurons to calcium overload while activated in a sustained manner during over-nutrition.

ZINC OXIDE NANOPARTICLES FOR ULTRAVIOLET PHOTODETECTION

2011 ◽  
Vol 20 (01) ◽  
pp. 183-194 ◽  
Author(s):  
SHAYLA SAWYER ◽  
LIQIAO QIN ◽  
CHRISTOPHER SHING
Keyword(s):  
Zinc Oxide ◽  
Gallium Nitride ◽  
Zno Nanoparticles ◽  
Green Emission ◽  
Synthesis Process ◽  
Voltage Response ◽  
Zno Nanoparticle ◽  
Linear Current ◽  
Current Voltage ◽  
Lift Off

Zinc Oxide ( ZnO ) nanoparticles were created by a top-down wet-chemistry synthesis process ( ZnO - A ) and then coated with polyvinyl-alcohol (PVA) ( ZnO - U ). In ZnO - U , strong UV emission was apparent while the parasitic green emission, which normally appears in ZnO suspensions, was suppressed. A standard lift-off process via e-beam lithography was used to fabricate a detector by evaporating Aluminum ( Al ) as ohmic electrodes on the ZnO nanoparticle film. Photoconductivity experiments showed that linear current-voltage response were achieved and the ZnO - U nanoparticles based detector had a ratio of UV photo-generated current more than 5 times better than that of the ZnO - A based detector. In addition, non-linear current-voltage responses were observed when interdigitated finger Gold ( Au ) contacts were deposited on ZnO - U . The UV generated current to dark current ratios were between 4 and 7 orders of magnitude, showing better performance than the photodetector with Al contacts. ZnO - U were also deposited on Gallium Nitride ( GaN ) and Aluminum Gallium Nitride ( AlGaN ) substrates to create spectrally selective photodetectors. The responsivity of detector based on AlGaN is twice that of commercial UV enhanced Silicon photodiodes. These results confirmed that ZnO nanoparticles coating with PVA is a good material for small-signal, visible blind, and wavelength selective UV detection.

PKC-sensitive Cl- channels associated with ciliary epithelial homologue of pICln

1995 ◽  
Vol 268 (3) ◽  
pp. C572-C579 ◽  
Author(s):  
M. Coca-Prados ◽  
J. Anguita ◽  
M. L. Chalfant ◽  
M. M. Civan
Keyword(s):  
Northern Analysis ◽  
Transmembrane Conductance Regulator ◽  
Transmembrane Conductance ◽  
Current Voltage ◽  
Cl Channels ◽  
Polymerase Chain ◽  
Reaction Cloning ◽  
Cystic Fibrosis Transmembrane ◽  
Nonpigmented Ciliary Epithelial ◽  
Current Voltage Relationship

Swelling activates and protein kinase C (PKC) downregulates Cl- channels in cultured nonpigmented ciliary epithelial (NPE) cells. We now report that the PKC inhibitor staurosporine upregulates whole cell Cl- currents isosmotically. The kinetics and current-voltage relationship are similar to those of volume-activated Cl- channels of these cells. These properties are inconsistent with cloned ClC-0, ClC-1, ClC-2, and MDR1 channels but could reflect the cystic fibrosis transmembrane conductance regulator (CFTR) channel or the Cl- channel regulator pICln. CFTR mRNA was undetectable by Northern analysis of cultured NPE cells or ciliary body tissue. In contrast, a human pICln probe obtained by polymerase chain reaction cloning and showing 90% identity with the rat cDNA clone detected high levels of transcripts in NPE cells. The level was low in tissue, where the NPE message was diluted by RNA from other cells. We conclude that NPE cells display staurosporine-activated Cl- channels [gSt(Cl)] likely identical with the volume-activated channels. The same cells expressing gSt(Cl) transcribe mRNA for a novel homologue (pHCBICln) of pICln that may regulate Cl- transport into the aqueous humor.

Measurement of parameters of the superconducting non-insulated coils

2021 ◽  
pp. 41-46
Author(s):  
Evgeniy P. Krasnoperov ◽  
Valentin V. Guryev ◽  
Vasyli V. Sychugov ◽  
Dmitri S. Yashkin ◽  
Sergei V. Shavkin
Keyword(s):  
Normal State ◽  
Superconducting State ◽  
Critical Value ◽  
Magnetic Characteristics ◽  
Electrical Characteristics ◽  
Linear Current ◽  
Superconducting Coils ◽  
Current Voltage ◽  
Current Input

The electrical characteristics of superconducting coils with non-insulated windings are studied. The procedures for measuring the parameters of uninsulated superconducting windings are described. In particular, the inductance is measured by voltage with a linear current input at a given rate. Attention is focused on the impossibility of correctly determining the inductance in a winding with an uninsulated superconductor in a normal state. It is noted that in a superconducting state at currents below the critical value, the inductance of the winding is comparable to the inductance with an insulated wire. The results of measurements of inductance, radial resistance, static current-voltage and magnetic characteristics of two tape coils with non-insulated superconducting windings, one of which had a soldered connection, are presented. Conditions for measuring the parameters of non-insulated superconducting windings are formulated when they are compared with insulated windings.

scholarly journals TOK Homologue in Neurospora crassa: First Cloning and Functional Characterization of an Ion Channel in a Filamentous Fungus

2003 ◽  
Vol 2 (1) ◽  
pp. 181-190 ◽  
Author(s):  
Stephen K. Roberts
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
Neurospora Crassa ◽  
Filamentous Fungus ◽  
Single Channel ◽  
Functional Characterization ◽  
Reversal Potential ◽  
Channel Activity ◽  
Biophysical Properties ◽  
Patch Clamp Technique

ABSTRACT In contrast to animal and plant cells, very little is known of ion channel function in fungal physiology. The life cycle of most fungi depends on the “filamentous” polarized growth of hyphal cells; however, no ion channels have been cloned from filamentous fungi and comparatively few preliminary recordings of ion channel activity have been made. In an attempt to gain an insight into the role of ion channels in fungal hyphal physiology, a homolog of the yeast K+ channel (ScTOK1) was cloned from the filamentous fungus, Neurospora crassa. The patch clamp technique was used to investigate the biophysical properties of the N. crassa K+ channel (NcTOKA) after heterologous expression of NcTOKA in yeast. NcTOKA mediated mainly time-dependent outward whole-cell currents, and the reversal potential of these currents indicated that it conducted K+ efflux. NcTOKA channel gating was sensitive to extracellular K+ such that channel activation was dependent on the reversal potential for K+. However, expression of NcTOKA was able to overcome the K+ auxotrophy of a yeast mutant missing the K+ uptake transporters TRK1 and TRK2, suggesting that NcTOKA also mediated K+ influx. Consistent with this, close inspection of NcTOKA-mediated currents revealed small inward K+ currents at potentials negative of EK. NcTOKA single-channel activity was characterized by rapid flickering between the open and closed states with a unitary conductance of 16 pS. NcTOKA was effectively blocked by extracellular Ca2+, verapamil, quinine, and TEA+ but was insensitive to Cs+, 4-aminopyridine, and glibenclamide. The physiological significance of NcTOKA is discussed in the context of its biophysical properties.

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