scholarly journals Calcium current activated upon hyperpolarization of Paramecium tetraurelia.

1992 ◽  
Vol 100 (2) ◽  
pp. 233-251 ◽  
Author(s):  
R R Preston ◽  
Y Saimi ◽  
C Kung
Keyword(s):  
Dissociation Constant ◽  
Calcium Current ◽  
Divalent Cations ◽  
Dependent Manner ◽  
Paramecium Tetraurelia ◽  
Apparent Dissociation Constant ◽  
Voltage Dependent ◽  
Effective Valence ◽  
The One ◽  
K Currents

Hyperpolarization of Paramecium tetraurelia under conditions where K+ currents are suppressed elicits an inward current that activates rapidly toward a peak at 25-80 ms and decays thereafter. This peak current (Ihyp) is not affected by removing Cl ions from the microelectrodes used to clamp membrane potential, or by changing extracellular Cl- concentration, but is lost upon removing extracellular Ca2+. Ihyp is also lost upon replacing extracellular Ca2+ with equimolar concentrations of Ba2+, Co2+, Mg2+, Mn2+, or Sr2+, suggesting that the permeability mechanism that mediates Ihyp is highly selective for Ca2+. Divalent cations also inhibit Ihyp when introduced extracellularly, in a concentration- and voltage-dependent manner. Ba2+ inhibits Ihyp with an apparent dissociation constant of 81 microM at -110 mV, and with an effective valence of 0.42. Ihyp is also inhibited reversibly by amiloride, with a dissociation constant of 0.4 mM. Ihyp is not affected significantly by changes in extracellular Na+, K+, or H+ concentration, or by EGTA injection. Also, it is unaffected by manipulations or mutations that suppress the depolarization-activated Ca2+ current or the various Ca(2+)-dependent currents of Paramecium. We suggest that Ihyp is mediated by a novel, hyperpolarization-activated calcium conductance that is distinct from the one activated by depolarization.

scholarly journals Batrachotoxin-modified sodium channels from squid optic nerve in planar bilayers. Ion conduction and gating properties.

1989 ◽  
Vol 93 (1) ◽  
pp. 23-41 ◽  
Author(s):  
M I Behrens ◽  
A Oberhauser ◽  
F Bezanilla ◽  
R Latorre
Keyword(s):  
Optic Nerve ◽  
Dissociation Constant ◽  
Sodium Channels ◽  
Single Channel ◽  
Dependent Manner ◽  
Channel Conductance ◽  
Apparent Dissociation Constant ◽  
Planar Bilayers ◽  
Voltage Range ◽  
Voltage Dependent

Squid optic nerve sodium channels were characterized in planar bilayers in the presence of batrachotoxin (BTX). The channel exhibits a conductance of 20 pS in symmetrical 200 mM NaCl and behaves as a sodium electrode. The single-channel conductance saturates with increasing the concentration of sodium and the channel conductance vs. sodium concentration relation is well described by a simple rectangular hyperbola. The apparent dissociation constant of the channel for sodium is 11 mM and the maximal conductance is 23 pS. The selectivity determined from reversal potentials obtained in mixed ionic conditions is Na+ approximately Li+ greater than K+ greater than Rb+ greater than Cs+. Calcium blocks the channel in a voltage-dependent manner. Analysis of single-channel membranes showed that the probability of being open (Po) vs. voltage relation is sigmoidal with a value of 0.5 between -90 and -100 mV. The fitting of Po requires at least two closed and one open state. The apparent gating charge required to move through the whole transmembrane voltage during the closed-open transition is four to five electronic charges per channel. Distribution of open and closed times are well described by single exponentials in most of the voltage range tested and mean open and mean closed times are voltage dependent. The number of charges associated with channel closing is 1.6 electronic charges per channel. Tetrodotoxin blocked the BTX-modified channel being the blockade favored by negative voltages. The apparent dissociation constant at zero potential is 16 nM. We concluded that sodium channels from the squid optic nerve are similar to other BTX-modified channels reconstituted in bilayers and to the BTX-modified sodium channel detected in the squid giant axon.

scholarly journals Ion permeation and block of the gating pore in the voltage sensor of NaV1.4 channels with hypokalemic periodic paralysis mutations

2010 ◽  
Vol 136 (2) ◽  
pp. 225-236 ◽  
Author(s):  
Stanislav Sokolov ◽  
Todd Scheuer ◽  
William A. Catterall
Keyword(s):  
Divalent Cations ◽  
Periodic Paralysis ◽  
Voltage Sensor ◽  
Dependent Manner ◽  
Hypokalemic Periodic Paralysis ◽  
Independent Manner ◽  
Apparent Dissociation Constant ◽  
Voltage Dependent ◽  
Arginine Residues ◽  
Trivalent Cations

Hypokalemic periodic paralysis and normokalemic periodic paralysis are caused by mutations of the gating charge–carrying arginine residues in skeletal muscle NaV1.4 channels, which induce gating pore current through the mutant voltage sensor domains. Inward sodium currents through the gating pore of mutant R666G are only ∼1% of central pore current, but substitution of guanidine for sodium in the extracellular solution increases their size by 13- ± 2-fold. Ethylguanidine is permeant through the R666G gating pore at physiological membrane potentials but blocks the gating pore at hyperpolarized potentials. Guanidine is also highly permeant through the proton-selective gating pore formed by the mutant R666H. Gating pore current conducted by the R666G mutant is blocked by divalent cations such as Ba2+ and Zn2+ in a voltage-dependent manner. The affinity for voltage-dependent block of gating pore current by Ba2+ and Zn2+ is increased at more negative holding potentials. The apparent dissociation constant (Kd) values for Zn2+ block for test pulses to −160 mV are 650 ± 150 µM, 360 ± 70 µM, and 95.6 ± 11 µM at holding potentials of 0 mV, −80 mV, and −120 mV, respectively. Gating pore current is blocked by trivalent cations, but in a nearly voltage-independent manner, with an apparent Kd for Gd3+ of 238 ± 14 µM at −80 mV. To test whether these periodic paralyses might be treated by blocking gating pore current, we screened several aromatic and aliphatic guanidine derivatives and found that 1-(2,4-xylyl)guanidinium can block gating pore current in the millimolar concentration range without affecting normal NaV1.4 channel function. Together, our results demonstrate unique permeability of guanidine through NaV1.4 gating pores, define voltage-dependent and voltage-independent block by divalent and trivalent cations, respectively, and provide initial support for the concept that guanidine-based gating pore blockers could be therapeutically useful.

scholarly journals Ion conductance and selectivity of single calcium-activated potassium channels in cultured rat muscle.

1984 ◽  
Vol 84 (1) ◽  
pp. 1-23 ◽  
Author(s):  
A L Blatz ◽  
K L Magleby
Keyword(s):  
Dissociation Constant ◽  
Single Channel ◽  
Reversal Potential ◽  
Ion Concentration ◽  
Dependent Manner ◽  
Apparent Dissociation Constant ◽  
Rat Muscle ◽  
Voltage Dependent ◽  
Single Channel Currents ◽  
Channel Currents

The conductance and selectivity of the Ca-activated K channel in cultured rat muscle was studied. Shifts in the reversal potential of single channel currents when various cations were substituted for Ki+ were used with the Goldman-Hodgkin-Katz equation to calculate relative permeabilities. The selectivity was Tl+ greater than K+ greater than Rb+ greater than NH4+, with permeability ratios of 1.2, 1.0, 0.67, and 0.11. Na+, Li+, and Cs+ were not measurably permeant, with permeabilities less than 0.05 that of K+. Currents with the various ions were typically less than expected on the basis of the permeability ratios, which suggests that the movement of an ion through the channel was not independent of the other ions present. For a fixed activity of Ko+ (77 mM), plots of single channel conductance vs. activity of Ki+ were described by a two-barrier model with a single saturable site. This observation, plus the finding that the permeability ratios of Rb+ and NH+4 to K+ did not change with ion concentration, is consistent with a channel that can contain a maximum of one ion at any time. The empirically determined dissociation constant for the single saturable site was 100 mM, and the maximum calculated conductance for symmetrical solutions of K+ was 640 pS. TEAi+ (tetraethylammonium ion) reduced single channel current amplitude in a voltage-dependent manner. This effect was accounted for by assuming voltage-dependent block by TEA+ (apparent dissociation constant of 60 mM at 0 mV) at a site located 26% of the distance across the membrane potential, starting at the inner side. TEAo+ was much more effective in reducing single channel currents, with an apparent dissociation constant of approximately 0.3 mM.

scholarly journals Charybdotoxin selectively blocks small Ca-activated K channels in Aplysia neurons.

1987 ◽  
Vol 90 (1) ◽  
pp. 27-47 ◽  
Author(s):  
A Hermann ◽  
C Erxleben
Keyword(s):  
Single Channel ◽  
Delayed Rectifier ◽  
Pacemaker Activity ◽  
Dependent Manner ◽  
Open Probability ◽  
Apparent Dissociation Constant ◽  
External Application ◽  
K Channels ◽  
Voltage Dependent ◽  
K Current

The action of charybdotoxin (ChTX), a peptide component isolated from the venom of the scorpion Leiurus quinquestriatus, was investigated on membrane currents of identified neurons from the marine mollusk, Aplysia californica. Macroscopic current recordings showed that the external application of ChTX blocks the Ca-activated K current in a dose- and voltage-dependent manner. The apparent dissociation constant is 30 nM at V = -30 mV and increases e-fold for a +50- to +70-mV change in membrane potential, which indicates that the toxin molecule is sensitive to approximately 35% of the transmembrane electric field. The toxin is bound to the receptor with a 1:1 stoichiometry and its effect is reversible after washout. The toxin also suppresses the membrane leakage conductance and a resting K conductance activated by internal Ca ions. The toxin has no significant effect on the inward Na or Ca currents, the transient K current, or the delayed rectifier K current. Records from Ca-activated K channels revealed a single channel conductance of 35 +/- 5 pS at V = 0 mV in asymmetrical K solution. The channel open probability increased with the internal Ca concentration and with membrane voltage. The K channels were blocked by submillimolar concentrations of tetraethylammonium ions and by nanomolar concentrations of ChTX, but were not blocked by 4-aminopyridine if applied externally on outside-out patches. From the effects of ChTX on K current and on bursting pacemaker activity, it is concluded that the termination of bursts is in part controlled by a Ca-activated K conductance.

scholarly journals Open Channel Block by Ca2+ Underlies the Voltage Dependence of Drosophila TRPL Channel

2006 ◽  
Vol 129 (1) ◽  
pp. 17-28 ◽  
Author(s):  
Moshe Parnas ◽  
Ben Katz ◽  
Baruch Minke
Keyword(s):  
Open Channel ◽  
Transient Receptor Potential ◽  
Voltage Dependence ◽  
Trp Channels ◽  
Divalent Cations ◽  
Dependent Manner ◽  
Channel Block ◽  
Open Channel Block ◽  
Gating Mechanism ◽  
Voltage Dependent

The light-activated channels of Drosophila photoreceptors transient receptor potential (TRP) and TRP-like (TRPL) show voltage-dependent conductance during illumination. Recent studies implied that mammalian members of the TRP family, which belong to the TRPV and TRPM subfamilies, are intrinsically voltage-gated channels. However, it is unclear whether the Drosophila TRPs, which belong to the TRPC subfamily, share the same voltage-dependent gating mechanism. Exploring the voltage dependence of Drosophila TRPL expressed in S2 cells, we found that the voltage dependence of this channel is not an intrinsic property since it became linear upon removal of divalent cations. We further found that Ca2+ blocked TRPL in a voltage-dependent manner by an open channel block mechanism, which determines the frequency of channel openings and constitutes the sole parameter that underlies its voltage dependence. Whole cell recordings from a Drosophila mutant expressing only TRPL indicated that Ca2+ block also accounts for the voltage dependence of the native TRPL channels. The open channel block by Ca2+ that we characterized is a useful mechanism to improve the signal to noise ratio of the response to intense light when virtually all the large conductance TRPL channels are blocked and only the low conductance TRP channels with lower Ca2+ affinity are active.

Substance P potentiates calcium channel modulation by somatostatin in chick sympathetic ganglia

1994 ◽  
Vol 72 (6) ◽  
pp. 2683-2690 ◽  
Author(s):  
A. Golard ◽  
L. Role ◽  
S. A. Siegelbaum
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Substance P ◽  
Dose Response ◽  
Calcium Current ◽  
Response Curve ◽  
Dose Response Curve ◽  
Dependent Manner ◽  
Kinase C ◽  
Voltage Dependent

1. The whole cell patch clamp was used to measure calcium current in isolated chick sympathetic ganglion neurons. Previous results showed that somatostatin inhibits calcium currents (ICa) in a voltage-dependent manner. The effect of somatostatin rapidly desensitizes. In addition, the action of somatostatin on the calcium current is inhibited by activation of protein kinase C (PKC). Because substance P (SP) has been shown to activate PKC in the chick sympathetic neurons, we here test the effects of SP on the calcium current and on the modulatory action of somatostatin. 2. At a concentration of 1 microM, SP had small, variable effects on ICa. 3. SP in the presence of guanosine 5'-triphosphate-gamma-S, or at higher concentrations (10 microM), inhibited ICa in a voltage-dependent manner, similar to the action of somatostatin. 4. Rather than inhibiting the action of somatostatin, SP (1 microM) potentiated the response to somatostatin. This effect of SP was only observed after the response to somatostatin had partially desensitized. SP had no effect on nondesensitized responses to somatostatin. 5. Desensitization of the somatostatin response involved a shift in its dose-response curve toward higher somatostatin concentrations as well as a decrease in the maximal response. SP appears to counteract the shift of the dose-response curve selectively. 6. The potentiation of the somatostatin response by SP is blocked by 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), but not by Calphostin C, Compound 5, k252a, protein kinase C (PKC)19-36, or adenylyl-imidodiphosphate (AMP-PNP), suggesting that phosphorylation is not involved and that the H-7 action does not depend on kinase inhibition.(ABSTRACT TRUNCATED AT 250 WORDS)

Permeation and block of dopamine-modulated potassium channels on rat striatal neurons by cesium and barium ions

1996 ◽  
Vol 76 (3) ◽  
pp. 1413-1422 ◽  
Author(s):  
Y. J. Lin ◽  
G. J. Greif ◽  
J. E. Freedman
Keyword(s):  
Dissociation Constant ◽  
Single Channel ◽  
Reversal Potential ◽  
Negative Slope ◽  
K Channel ◽  
Striatal Neurons ◽  
Apparent Dissociation Constant ◽  
Voltage Dependent ◽  
Single Channel Currents ◽  
Inwardly Rectifying

1. In cell-attached patch-clamp recordings from freshly dissociated rat caudate-putamen neurons, an 85-pS inwardly rectifying K+ channel, which was previously found to be modulated by D2-like dopamine receptors, was blocked by externally applied BaCl2 or CsCl. 2. At concentrations between 100 and 500 microM, Ba2+ blockade was voltage dependent, with a greater block at hyperpolarized voltages, in a manner consistent with blockade of the channel pore. Single-channel currents were flickery, with intervening periods of more complete blockade, and block appeared to be time dependent, with an estimated electrical distance of 0.24 and an apparent dissociation constant of 205 microM at 0 mV. 3. At concentrations between 1 and 3 mM, Cs+ blockade was similarly voltage dependent, but without periods of longer blockade, with an electrical distance of 0.81 and an apparent dissociation constant of 625 microM at 0 mV. Cs+ could also permeate this channel at voltages near the K+ reversal potential. The current-voltage relationship displayed an anomalous negative slope conductance, in a manner inconsistent with a single-ion pore. 4. Smaller-conductance, dopamine-insensitive channels were blocked more potently by both Ba2+ and Cs+ than was the 85-pS channel, reflecting differences between inwardly rectifying K+ channels mediating resting conductance and those mediating dopamine receptor responses in striatal neurons.

Separation and modulation of calcium currents in bullfrog sympathetic neurons

1992 ◽  
Vol 70 (S1) ◽  
pp. S56-S63 ◽  
Author(s):  
Stephen W. Jones ◽  
Keith S. Elmslie
Keyword(s):  
G Protein ◽  
Calcium Current ◽  
Sympathetic Neurons ◽  
Calcium Currents ◽  
Dependent Manner ◽  
Activation Kinetics ◽  
Channel Opening ◽  
Voltage Dependent ◽  
Rapid Activation ◽  
Kinetics Of

The calcium current of frog sympathetic neurons has relatively rapid activation kinetics (τ < 3 ms) in response to changes in voltage. Pharmacologically, the current is blocked ~90% by ω-conotoxin, but < 10% by dihydropyridine antagonists. This suggests that nearly all of the current is N type. However, inactivation is slow and incomplete even for depolarizations lasting > 1 s, consistent with recent evidence that N-type channels do not always inactivate rapidly. The calcium current is partially inhibited via receptors for acetylcholine, luteinizing hormone releasing hormone, substance P, ATP, and norepinephrine. These effects are mimicked by internal dialysis with GTP-γ-S, suggesting involvement of a G protein. The transmitters affect the activation kinetics of the calcium current in a voltage-dependent manner, which can be modeled as a reversible shift of some channels to "reluctant" states in which strong depolarization is needed to produce channel opening. The effects of transmitters develop and recover with t½ ~ 1–2 s, so if a second messenger is involved in receptor – calcium channel coupling, it must act rapidly.Key words: norepinephrine, ω-conotoxin, dihydropyridine, inactivation, G protein.

scholarly journals Polyvalent Cations Constitute the Voltage Gating Particle in Human Connexin37 Hemichannels

2004 ◽  
Vol 124 (5) ◽  
pp. 587-603 ◽  
Author(s):  
Michael C. Puljung ◽  
Viviana M. Berthoud ◽  
Eric C. Beyer ◽  
Dorothy A. Hanck
Keyword(s):  
Divalent Cations ◽  
Voltage Gating ◽  
Bathing Solution ◽  
Local Concentration ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Current Decay ◽  
Gap Junction Channel ◽  
Polyvalent Cations ◽  
Voltage Dependent

Connexins oligomerize to form intercellular channels that gate in response to voltage and chemical agents such as divalent cations. Historically, these are believed to be two independent processes. Here, data for human connexin37 (hCx37) hemichannels indicate that voltage gating can be explained as block/unblock without the necessity for an independent voltage gate. hCx37 hemichannels closed at negative potentials and opened in a time-dependent fashion at positive potentials. In the absence of polyvalent cations, however, the channels were open at relatively negative potentials, passing current linearly with respect to voltage. Current at negative potentials could be inhibited in a concentration-dependent manner by the addition of polyvalent cations to the bathing solution. Inhibition could be explained as voltage-dependent block of hCx37, with the field acting directly on polyvalent cations, driving them through the pore to an intracellular site. At positive potentials, in the presence of polyvalent cations, the field favored polyvalent efflux from the intracellular blocking site, allowing current flow. The rate of appearance of current depended on the species and valence of the polyvalent cation in the bathing solution. The rate of current decay upon repolarization depended on the concentration of polyvalent cations in the bathing solution, consistent with deactivation by polyvalent block, and was rapid (time constants of tens of milliseconds), implying a high local concentration of polyvalents in or near the channel pore. Sustained depolarization slowed deactivation in a flux-dependent, voltage- and time-independent fashion. The model for hCx37 voltage gating as polyvalent block/unblock can be expanded to account for observations in the literature regarding hCx37 gap junction channel behavior.

Three distinct G protein pathways mediate inhibition of neuronal calcium current by bradykinin

1996 ◽  
Vol 76 (5) ◽  
pp. 3559-3562 ◽  
Author(s):  
M. A. Wilk-Blaszczak ◽  
W. D. Singer ◽  
F. Belardetti
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Pertussis Toxin ◽  
Calcium Current ◽  
Suppressive Effect ◽  
Calcium Currents ◽  
Tetraacetic Acid ◽  
Combined Application ◽  
Voltage Dependent ◽  
K Currents

1. In NG108-15 cells dialyzed with 10 mM ethylene glycolbis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) or bis (o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA), bradykinin (BK) selectively inhibited the N-type calcium current. This effect of BK was blocked by an antibody directed against the G protein G13. Thus under these conditions G13 mediates the inhibition of voltage-dependent calcium current (ICa, V) by BK. In contrast, activation of K+ currents by BK is mediated by Gq/11. BK also couples to Gi2. 2. We now examine the involvement of G proteins in the inhibition of ICa, V by BK when NG108-15 cells are dialyzed with 1 mM BAPTA. Under these conditions, BK inhibited both the N- and L-type, but not the T-type, calcium currents. Intracellular application of anti-G13 antibody did not suppress the response to BK. Applications of either anti-Gq/11 antibody or pertussis toxin (PTX, to block Gi2) were similarly ineffective. Even combined application of anti-Gq/11 and -G13 antibodies, or PTX together with either antibody, did not block inhibition of ICa, V by BK. However, the combination of both antibodies with PTX blocked the response to BK in low BAPTA. In conclusion, both Gq/11 and a PTX-sensitive G protein (presumably Gi2), together with G13, are involved in the inhibition of ICa, V by BK. 3. Gq/11 inhibited only the L-type calcium current, whereas the PTX-sensitive G protein inhibited both the N- and L-type calcium currents. 4. The BAPTA dependence of the Gq/11 and PTX-sensitive inhibitions may reflect a Ca2+ requirement of the pathway(s) acting on the L current and/or a direct suppressive effect of BAPTA.

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