scholarly journals The pacemaker current in cardiac Purkinje myocytes.

1995 ◽  
Vol 106 (3) ◽  
pp. 559-578 ◽  
Author(s):  
M Vassalle ◽  
H Yu ◽  
I S Cohen
Keyword(s):  
Reversal Potential ◽  
Time Dependent ◽  
Tyrode Solution ◽  
Voltage Range ◽  
Inward Current ◽  
Whole Cell Patch Clamp ◽  
Diastolic Depolarization ◽  
Cardiac Purkinje Fibers ◽  
K Current ◽  
K Depletion

It is generally assumed that in cardiac Purkinje fibers the hyperpolarization activated inward current i(f) underlies the pacemaker potential. Because some findings are at odds with this interpretation, we used the whole cell patch clamp method to study the currents in the voltage range of diastolic depolarization in single canine Purkinje myocytes, a preparation where many confounding limitations can be avoided. In Tyrode solution ([K+]o = 5.4 mM), hyperpolarizing steps from Vh = -50 mV resulted in a time-dependent inwardly increasing current in the voltage range of diastolic depolarization. This time-dependent current (iKdd) appeared around -60 mV and reversed near EK. Small superimposed hyperpolarizing steps (5 mV) applied during the voltage clamp step showed that the slope conductance decreases during the development of this time-dependent current. Decreasing [K+]o from 5.4 to 2.7 mM shifted the reversal potential to a more negative value, near the corresponding EK. Increasing [K+]o to 10.8 mM almost abolished iKdd. Cs+ (2 mM) markedly reduced or blocked the time-dependent current at potentials positive and negative to EK. Ba2+ (4 mM) abolished the time-dependent current in its usual range of potentials and unmasked another time-dependent current (presumably i(f)) with a threshold of approximately -90 mV (> 20 mV negative to that of the time-dependent current in Tyrode solution). During more negative steps, i(f) increased in size and did not reverse. During i(f) the slope conductance measured with small (8-10 mV) superimposed clamp steps increased. High [K+]o (10.8 mM) markedly increased and Cs+ (2 mM) blocked i(f). We conclude that: (a) in the absence of Ba2+, a time-dependent current does reverse near EK and its reversal is unrelated to K+ depletion; (b) the slope conductance of that time-dependent current decreases in the absence of K+ depletion at potentials positive to EK where inactivation of iK1 is unlikely to occur. (c) Ba2+ blocks this time-dependent current and unmasks another time-dependent current (i(f)) with a more negative (> 20 mV) threshold and no reversal at more negative values; (d) Cs+ blocks both time-dependent currents recorded in the absence and presence of Ba2+. The data suggest that in the diastolic range of potentials in Purkinje myocytes there is a voltage- and time-dependent K+ current (iKdd) that can be separated from the hyperpolarization-activated inward current i(f).

scholarly journals A time-dependent and voltage-sensitive K+ current in single cells from frog atrium.

1986 ◽  
Vol 88 (6) ◽  
pp. 739-755 ◽  
Author(s):  
M A Simmons ◽  
T Creazzo ◽  
H C Hartzell
Keyword(s):  
Single Cells ◽  
Quantitative Description ◽  
Reversal Potential ◽  
Outward Current ◽  
Time Dependent ◽  
Patch Clamp Technique ◽  
Whole Cell Patch Clamp ◽  
K Current ◽  
And Shifts ◽  
Frog Atrium

A quantitative description of the time-dependent and voltage-sensitive outward currents in heart has been hampered by the complications inherent to the multicellular preparations previously used. We have used the whole-cell patch-clamp technique to record the delayed outward K+ current, IK, in single cells dissociated from frog atrium. Na+ currents were blocked with tetrodotoxin and Ca2+ currents with Mn2+ or Cd2+. After depolarizations from -50 mV to potentials positive to -30 mV, a time-dependent outward current was observed. This current has been characterized according to its steady state activation, kinetics, and ion transfer function. The current is well described as a single Hodgkin-Huxley conductance. The deactivation of the current is a single exponential. Activation of the current is sigmoid and is fitted well by raising the activation variable to the second power. The reversal potential of IK is near EK and shifts by 57 mV/10-fold change in [K+]o. This suggests that the current is carried selectively by K ions. The threshold for activation is near -30 mV. IK is maximally activated positive to +20 mV and shows no inactivation. The fully activated current-voltage relationship is linear between -110 and +50 mV. Neither Ba2+ (250 microM) nor Cd2+ (100 microM) affects IK.

scholarly journals Two components of cardiac delayed rectifier K+ current. Differential sensitivity to block by class III antiarrhythmic agents.

1990 ◽  
Vol 96 (1) ◽  
pp. 195-215 ◽  
Author(s):  
M C Sanguinetti ◽  
N K Jurkiewicz
Keyword(s):  
Antiarrhythmic Agent ◽  
Reversal Potential ◽  
Differential Sensitivity ◽  
Inward Rectification ◽  
Delayed Rectifier ◽  
Antiarrhythmic Agents ◽  
Class Iii ◽  
Activation Curve ◽  
Voltage Range ◽  
K Current

An envelope of tails test was used to show that the delayed rectifier K+ current (IK) of guinea pig ventricular myocytes results from the activation of two outward K+ currents. One current was specifically blocked by the benzenesulfonamide antiarrhythmic agent, E-4031 (IC50 = 397 nM). The drug-sensitive current, "IKr" exhibits prominent rectification and activates very rapidly relative to the slowly activating drug-insensitive current, "IKs." IKs was characterized by a delayed onset of activation that occurs over a voltage range typical of the classically described cardiac IK. Fully activated IKs, measured as tail current after 7.5-s test pulses, was 11.4 times larger than the fully activated IKr. IKr was also blocked by d-sotalol (100 microM), a less potent benzenesulfonamide Class III antiarrhythmic agent. The activation curve of IKr had a steep slope (+7.5 mV) and a negative half-point (-21.5 mV) relative to the activation curve of IKs (slope = +12.7 mV, half-point = +15.7 mV). The reversal potential (Erev) of IKr (-93 mV) was similar to EK (-94 mV for [K+]o = 4 mM), whereas Erev of IKs was -77 mV. The time constants for activation and deactivation of IKr made up a bell-shaped function of membrane potential, peaking between -30 and -40 mV (170 ms). The slope conductance of the linear portion of the fully activated IKr-V relation was 22.5 S/F. Inward rectification of this relation occurred at potentials greater than -50 mV, resulting in a voltage-dependent decrease in peak IKr at test potentials greater than 0 mV. Peak IKr at 0 mV averaged 0.8 pA/pF (n = 21). Although the magnitude of IKr was small relative to fully activated IKs, the two currents were of similar magnitude when measured during a relatively short pulse protocol (225 ms) at membrane potentials (-20 to +20 mV) typical of the plateau phase of cardiac action potentials.

Multiple Effects of Serotonin and Acetylcholine on Hyperpolarization-Activated Inward Current in Locomotor Activity-Related Neurons in Cfos-EGFP Mice

2010 ◽  
Vol 104 (1) ◽  
pp. 366-381 ◽  
Author(s):  
Yue Dai ◽  
Larry M. Jordan
Keyword(s):  
Spinal Cord ◽  
Motor System ◽  
Reversal Potential ◽  
Rhythmic Activity ◽  
Maximal Conductance ◽  
Inward Current ◽  
Spinal Interneurons ◽  
Whole Cell Patch Clamp ◽  
Activation Rate ◽  
Maximal Activation

Hyperpolarization-activated inward current ( Ih) has been shown to be involved in production of bursting during various forms of rhythmic activity. However, details of Ih in spinal interneurons related to locomotion remain unknown. Using Cfos-EGFP transgenic mice (P6–P12) we are able to target the spinal interneurons activated by locomotion. Following a locomotor task, whole cell patch-clamp recordings were obtained from ventral EGFP+ neurons in spinal cord slices (T13–L4, 200–250 μm). Ih was found in 51% of EGFP+ neurons ( n = 149) with almost even distribution in lamina VII (51%), VIII (47%), and X (55%). Ih could be blocked by ZD7288 (10–20 μM) or cesium (1–1.5 mM) but was insensitive to barium (2–2.5 mM). Ih activated at −80.1 ± 9.2 mV with half-maximal activation −95.5 ± 13.3 mV, activation rate 10.0 ± 3.2 mV, time constant 745 ± 501 ms, maximal conductance 1.0 ± 0.7 nS, and reversal potential −34.3 ± 3.6 mV. 5-HT (15–20 μM) and ACh (20–30 μM) produced variable effects on Ih. 5-HT increased Ih in 43% of EGFP+ neurons ( n = 37), decreased Ih in 24%, and had no effect on Ih in 33% of the neurons. ACh decreased Ih in 67% of EGFP+ neurons ( n = 18) with unchanged Ih in 33% of the neurons. This study characterizes the Ih in locomotor-related interneurons and is the first to demonstrate the variable effects of 5-HT and ACh on Ih in rodent spinal interneurons. The finding of 5-HT and ACh-induced reduction of Ih in EGFP+ neurons suggests a novel mechanism that the motor system could use to limit the participation of certain neurons in locomotion.

Ca2+-activated Cl− current in cultured myenteric neurons from murine proximal colon

2003 ◽  
Vol 284 (4) ◽  
pp. C839-C847 ◽  
Author(s):  
Sok Han Kang ◽  
Pieter Vanden Berghe ◽  
Terence K. Smith
Keyword(s):  
Membrane Potential ◽  
Channel Blocker ◽  
Reversal Potential ◽  
Outward Current ◽  
Proximal Colon ◽  
Time Dependent ◽  
Equilibrium Potential ◽  
Resting Membrane Potential ◽  
Myenteric Neurons ◽  
Whole Cell Patch Clamp

Whole cell patch-clamp recordings were made from cultured myenteric neurons taken from murine proximal colon. The micropipette contained Cs+ to remove K+ currents. Depolarization elicited a slowly activating time-dependent outward current ( I tdo), whereas repolarization was followed by a slowly deactivating tail current ( I tail). I tdo and I tail were present in ∼70% of neurons. We identified these currents as Cl− currents ( I Cl), because changing the transmembrane Cl− gradient altered the measured reversal potential ( E rev) of both I tdo and I tail with that for I tailshifted close to the calculated Cl− equilibrium potential ( E Cl). I Cl are Ca2+-activated Cl− current [ I Cl(Ca)] because they were Ca2+dependent. E Cl, which was measured from the E rev of I Cl(Ca) using a gramicidin perforated patch, was −33 mV. This value is more positive than the resting membrane potential (−56.3 ± 2.7 mV), suggesting myenteric neurons accumulate intracellular Cl−. ω-Conotoxin GIVA [0.3 μM; N-type Ca2+ channel blocker] and niflumic acid [10 μM; known I Cl(Ca) blocker], decreased the I Cl(Ca). In conclusion, these neurons have I Cl(Ca) that are activated by Ca2+entry through N-type Ca2+ channels. These currents likely regulate postspike frequency adaptation.

A Novel Ca2+ Influx Pathway in Mammalian Primary Sensory Neurons Is Activated by Caffeine

2001 ◽  
Vol 86 (1) ◽  
pp. 190-196 ◽  
Author(s):  
Robert E. Hoesch ◽  
Daniel Weinreich ◽  
Joseph P. Y. Kao
Keyword(s):  
Sensory Neurons ◽  
Ryanodine Receptors ◽  
Reversal Potential ◽  
Significant Rise ◽  
Adult Rabbit ◽  
Pharmacological Agents ◽  
Inward Current ◽  
Whole Cell Patch Clamp ◽  
Intracellular Ca ◽  
Ganglion Neurons

Single-cell microfluorimetry and electrophysiology techniques were used to identify and characterize a novel Ca2+ influx pathway in adult rabbit vagal sensory neurons. Acutely dissociated nodose ganglion neurons (NGNs) exhibit robust Ca2+-induced Ca2+ release (CICR) that can be triggered by 10 mM caffeine, the classic agonist of CICR. A caffeine-induced increase in cytosolic-free Ca2+ concentration ([Ca2+]i) is considered diagnostic evidence of the existence of CICR. However, when CICR was disabled through depletion of intracellular Ca2+stores or pharmacological blockade of intracellular Ca2+ release channels (ryanodine receptors), caffeine still elicited a significant rise in [Ca2+]i in ∼50% of NGNs. The same response was not elicited by pharmacological agents that elevate cyclic nucleotide concentrations. Moreover, extracellular Ca2+ was obligatory for such caffeine-induced [Ca2+]i rises in this population of NGNs, suggesting that Ca2+ influx is responsible for this rise. Simultaneous microfluorimetry with whole cell patch-clamp studies showed that caffeine activates an inward current that temporally parallels the rise in [Ca2+]i. The inward current had a reversal potential of +8.1 ± 6.1 (SE) mV ( n = 4), a mean peak amplitude of −126 ± 24 pA ( n = 4) at E m = −50 mV, and a slope conductance of 1.43 ± 0.79 nS ( n= 4). Estimated EC50 values for caffeine-induced CICR and for caffeine-activated current were 1.5 and ∼0.6 mM, respectively. These results indicate that caffeine-induced rises in [Ca2+]i, in the presence of extracellular Ca2+, can no longer be interpreted as unequivocal diagnostic evidence for CICR in neurons. These results also indicate that sensory neurons possess a novel Ca2+ influx pathway.

Properties of potassium currents in Purkinje cells of failing human hearts

2002 ◽  
Vol 283 (6) ◽  
pp. H2495-H2503 ◽  
Author(s):  
Wei Han ◽  
Liming Zhang ◽  
Gernot Schram ◽  
Stanley Nattel
Keyword(s):  
Purkinje Cells ◽  
Ventricular Myocytes ◽  
Outward Current ◽  
Ionic Currents ◽  
Time Dependent ◽  
Transient Outward Current ◽  
Whole Cell Patch Clamp ◽  
Cardiac Purkinje Fibers ◽  
Cardiac Purkinje Cells ◽  
Hyperpolarization Activated Current

Cardiac Purkinje fibers play an important role in cardiac arrhythmias, but no information is available about ionic currents in human cardiac Purkinje cells (PCs). PCs and midmyocardial ventricular myocytes (VMs) were isolated from explanted human hearts. K+ currents were evaluated at 37°C with whole cell patch clamp. PCs had clear inward rectifier K+current ( I K1), with a density not significantly different from VMs between −110 and −20 mV. A Cs+-sensitive, time-dependent hyperpolarization-activated current was measurable negative to −60 mV. Transient outward current ( I to) density was smaller, but end pulse sustained current ( I sus) was larger, in PCs vs. VMs. I to recovery was substantially slower in PCs, leading to strong frequency dependence. Unlike VM I to, which was unaffected by 10 mM tetraethylammonium, Purkinje I to was strongly inhibited by tetraethylammonium, and Purkinje I to was 10-fold more sensitive to 4-aminopyridine than VM. PC I sus was also reduced strongly by 10 mM tetraethylammonium. In conclusion, human PCs demonstrate a prominent I K1, a time-dependent hyperpolarization-activated current, and an I towith pharmacological sensitivity and recovery kinetics different from those in the atrium or ventricle and compatible with a different molecular basis.

Channels involved in transient currents unmasked by removal of extracellular calcium in cardiac cells

2002 ◽  
Vol 282 (5) ◽  
pp. H1879-H1888 ◽  
Author(s):  
Regina Macianskiene ◽  
Francesco Moccia ◽  
Karin R. Sipido ◽  
Willem Flameng ◽  
Kanigula Mubagwa
Keyword(s):  
Ventricular Myocytes ◽  
Reversal Potential ◽  
Monovalent Cation ◽  
Cardiac Cells ◽  
Time Dependent ◽  
Patch Clamp Technique ◽  
Whole Cell Patch Clamp ◽  
Voltage Dependent ◽  
High Concentrations ◽  
Low Sensitivity

In cardiac cells that lack macroscopic transient outward K+ currents ( I to), the removal of extracellular Ca2+ can unmask “ I to-like” currents. With the use of pig ventricular myocytes and the whole cell patch-clamp technique, we examined the possibility that cation efflux via L-type Ca2+channels underlies these currents. Removal of extracellular Ca2+ and extracellular Mg2+ induced time-independent currents at all potentials and time-dependent currents at potentials greater than −50 mV. Either K+ or Cs+ could carry the time-dependent currents, with reversal potential of +8 mV with internal K+ and +34 mV with Cs+. Activation and inactivation were voltage dependent [Boltzmann distributions with potential of half-maximal value ( V 1/2) = −24 mV and slope = −9 mV for activation; V 1/2 = −58 mV and slope = 13 mV for inactivation]. The time-dependent currents were resistant to 4-aminopyridine and to DIDS but blocked by nifedipine at high concentrations (IC50 = 2 μM) as well as by verapamil and diltiazem. They could be increased by BAY K-8644 or by isoproterenol. We conclude that the I to-like currents are due to monovalent cation flow through L-type Ca2+ channels, which in pig myocytes show low sensitivity to nifedipine.

Mechanosensitive currents in putative aortic baroreceptor neurons in vitro

1995 ◽  
Vol 73 (5) ◽  
pp. 2094-2098 ◽  
Author(s):  
J. T. Cunningham ◽  
R. E. Wachtel ◽  
F. M. Abboud
Keyword(s):  
Reversal Potential ◽  
Tumor Cell Line ◽  
Mouse Tumor ◽  
Mechanosensitive Channels ◽  
Inward Current ◽  
Voltage Gated ◽  
Voltage Gated Calcium Channels ◽  
Whole Cell Patch Clamp ◽  
Nodose Ganglia

1. Whole cell patch-clamp experiments were conducted to determine whether rat aortic baroreceptor neurons contain mechano-sensitive conductances. 2. Putative aortic baroreceptor neurons in the nodose ganglia were identified by injecting DiI onto the adventitia of the aortic arch. Nodose ganglia neurons were dissociated after > or = 1 wk. A fluorescein-conjugated tetanus toxin fragment was used to confirm that the cells labeled with DiI in culture were neurons. 3. Hypoosmotic stretch significantly increased the conductance of DiI-labeled neurons (n = 19). The reversal potential of the response was -11 +/- 1 (SE) mV. 4. In experiments on unlabeled neurons, only 7 of 13 cells showed increases in conductance. BC3H1 cells, a mouse tumor cell line, showed no changes in conductance. 5. Gadolinium (20 microM), a putative blocker of mechanosensitive channels, prevented the increase in conductance produced by hypoosmolality in seven of seven labeled cells. Equimolar concentrations of lanthanum (n = 6) and omega-conotoxin GVIA (1 microM, n = 4), which block voltage-gated calcium channels, failed to significantly affect the inward current.

Intracellular pH modulates the activity of chloride channels in isolated lacrimal gland acinar cells

1995 ◽  
Vol 268 (3) ◽  
pp. C647-C650 ◽  
Author(s):  
K. Park ◽  
P. D. Brown
Keyword(s):  
Intracellular Ph ◽  
Lacrimal Gland ◽  
Chloride Channels ◽  
Reversal Potential ◽  
Acinar Cells ◽  
Secretory Activity ◽  
Time Dependent ◽  
Whole Cell ◽  
Whole Cell Patch Clamp ◽  
Cl Channels

The effects of intracellular pH (pHi) on Ca(2+)-activated Cl- currents in rat lacrimal gland acinar cells were examined. Cl- currents were recorded by conventional whole cell patch-clamp methods using K(+)-free and Na(+)-free solutions. pHi was varied by using electrode solutions with pH at 6.8, 7.3, or 7.8, and Ca2+ activity was buffered at 100 nM with ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. Increasing pH from 6.8 to 7.8 was found to increase whole cell currents. The currents observed exhibited time-dependent activation at depolarizing potentials and time-dependent inactivation at hyperpolarizing potentials (pH 7.8). This behavior is characteristic of Ca(2+)-activated Cl- channels in lacrimal gland cells. The selectivity of the current was examined at pH 7.8 by removing Cl- from the bath solution. This maneuver caused a positive shift in the reversal potential, as expected for a Cl(-)-selective current. Thus increasing pHi appears to activate Ca(2+)-activated Cl- channels. The possibility that an increase in pHi may help sustain Cl- channel activity during secretory activity is discussed.

scholarly journals Modulation of Calcium Channels in Human Erythroblasts by Erythropoietin

Blood ◽  
1997 ◽  
Vol 89 (1) ◽  
pp. 92-100 ◽  
Author(s):  
Joseph Y. Cheung ◽  
Xue-Qian Zhang ◽  
Krister Bokvist ◽  
Douglas L. Tillotson ◽  
Barbara A. Miller
Keyword(s):  
Single Channel ◽  
Reversal Potential ◽  
Erythropoietin Receptor ◽  
Single Channels ◽  
Open Probability ◽  
Whole Cell ◽  
Voltage Range ◽  
Whole Cell Patch Clamp ◽  
Open Time ◽  
High Doses

Abstract Erythropoietin (Epo) induces a dose-dependent increase in intracellular free Ca2+ ([Ca2+]i ) in human erythroblasts, which is dependent on extracellular Ca2+ and blocked by high doses of nifedipine or Ni2+. In addition, pretreatment of human erythroblasts with mouse antihuman erythropoietin receptor antibody but not mouse immunopure IgG blocked the Epo-induced [Ca2+]i increase, indicating the specificity of the Ca2+ response to Epo stimulation. In this study, the erythropoietin-regulated calcium channel was identified by single channel recordings. Use of conventional whole cell patch-clamp failed to detect Epo-induced whole cell Ca2+ current. To minimize washout of cytosolic constituents, we next used nystatin perforated patch, but did not find any Epo-induced whole cell Ca2+ current. Using Ba2+ (30 mmol/L) as charge carrier in cell-attached patches, we detected single channels with unitary conductance of 3.2 pS, reversal potential of +72 mV, and whose unitary current (at +10 mV) increased monotonically with increasing Ba2+ concentrations. Channel open probability did not appreciably change over the voltage range (−50 to +30 mV) tested. Epo (2 U/mL) increased both mean open time (from 4.27 ± 0.75 to 11.15 ± 1.80 ms) and open probability (from 0.26 ± 0.06 to 2.56 ± 0.59%) of this Ba2+-permeable channel. Our data strongly support the conclusion that the Epo-induced [Ca2+]i increase in human erythroblasts is mediated via Ca2+ entry through a voltage-independent Ca2+ channel.

Sign in / Sign up

Export Citation Format

Share Document