Temperature-dependence of L-type Ca(2+) channel current in atrial myocytes from rainbow trout

2000 ◽  
Vol 203 (18) ◽  
pp. 2771-2780 ◽  
Author(s):  
H.A. Shiels ◽  
M. Vornanen ◽  
A.P. Farrell
Keyword(s):  
Rainbow Trout ◽  
Temperature Dependence ◽  
Voltage Clamp ◽  
Pulse Voltage ◽  
Kinetic Properties ◽  
Temperature Dependency ◽  
Atrial Myocytes ◽  
Patch Clamp Technique ◽  
Channel Current ◽  
Series Of Experiments

Rainbow trout, Oncorhynchus mykiss, inhabit eurythermal environments and must therefore be able to cope with changes in environmental temperature. As ectotherms, their heart is required to maintain cardiac function over a range of ambient water temperatures. This raises important questions concerning the temperature-dependence of cardiac ion channel function in fish hearts, in particular, the channels involved in Ca(2+) transport. Thus, we studied the effects of acute, physiologically relevant temperature changes on the density and kinetics of the L-type Ca(2+) channel current (I(Ca)) in rainbow trout atrial myocytes using the whole-cell patch-clamp technique. Myocytes from fish acclimated to 14 degrees C were first tested at 14 degrees C, then at 21 degrees C and finally at 7 degrees C. Using a square-pulse voltage-clamp in the first series of experiments, the peak density of I(Ca) increased (Q(10)=1.9) as temperature was increased from 14 to 21 degrees C and decreased (Q(10)=2.1) as temperature was decreased from 14 to 7 degrees C. In contrast to current density, the charge carried by I(Ca) was inversely related to temperature as a result of changes in the kinetic properties of the channel; both the fast (tau(f)) and slow (tau(s)) components of inactivation were slower at 7 degrees C than at 14 and 21 degrees C. Action potentials were recorded at the three test temperatures and then used as voltage-clamp stimulus waveforms to reassess I(Ca) in a second series of experiments. While the temperature-dependency of I(Ca) was similar to that found with the square-pulse voltage-clamp, the charge carried by I(Ca) was temperature-independent. These results show that the temperature-dependency of I(Ca) in rainbow trout is in the lower range of that reported in mammals and, although this could have profound effects on Ca(2+) delivery to the myofilaments, the temperature-induced modifications in the action potential may help to maintain a fairly constant Ca(2+) delivery during an acute temperature change in rainbow trout.

Voltage-dependent calcium channels in ventricular cells of rainbow trout: effect of temperature changes in vitro

2000 ◽  
Vol 278 (6) ◽  
pp. R1524-R1534 ◽  
Author(s):  
Catherine S. Kim ◽  
Mary D. Coyne ◽  
Judith K. Gwathmey
Keyword(s):  
Rainbow Trout ◽  
Calcium Channels ◽  
Temperature Sensitivity ◽  
Calcium Currents ◽  
Temperature Dependency ◽  
Patch Clamp Technique ◽  
Ventricular Cells ◽  
Voltage Dependent ◽  
Voltage Dependent Calcium Channels

Voltage-dependent calcium channels (VDCC) in ventricular myocytes from rainbow trout ( Oncorhynchus mykiss) were investigated in vitro using the perforated patch-clamp technique, which maintains the integrity of the intracellular milieu. First, we characterized the current using barium as the charge carrier and established the doses of various pharmacological agents to use these agents in additional studies. Second, we examined the current at several physiological temperatures to determine temperature dependency. The calcium currents at 10°C (acclimation temperature) were identified as l-type calcium currents based on their kinetic behavior and response to various calcium channel agonists and antagonists. Myocytes were chilled (4°C) and warmed (18 and 22°C), and the response of VDCC to varying temperatures was observed. There was no significant dependency of the current amplitude and kinetics on temperature. Amplitude decreased 25–36% at 4°C (Q10 ∼1.89) and increased 18% at 18°C (Q10 ∼1.23) in control, Bay K8644 (Bay K)-, and forskolin-enhanced currents. The inactivation rates (τi) did not demonstrate a temperature sensitivity for the VDCC (Q10 1.23–1.92); Bay K treatment, however, increased temperature sensitivity of τi between 10 and 18°C (Q10 3.98). The low Q10 values for VDCC are consistent with a minimal temperature sensitivity of trout myocytes between 4 and 22°C. This low-temperature dependency may provide an important role for sarcolemmal calcium channels in adaptation to varying environmental temperatures in trout.

Na+/Ca2+-exchange activity regulates contraction and SR Ca2+ content in rainbow trout atrial myocytes

2000 ◽  
Vol 279 (5) ◽  
pp. R1856-R1864 ◽  
Author(s):  
Leif Hove-Madsen ◽  
Anna Llach ◽  
Lluis Tort
Keyword(s):  
Rainbow Trout ◽  
Sarcoplasmic Reticulum ◽  
Atrial Myocytes ◽  
Initial Value ◽  
Inhibitory Peptide ◽  
Patch Clamp Technique ◽  
Cell Shortening ◽  
Control Value ◽  
Patch Pipette ◽  
Exchange Activity

We have used the whole cell configuration of the patch-clamp technique to measure sarcolemmal Ca2+ transport by the Na+/Ca2+ exchanger (NCX) and its contribution to the activation and relaxation of contraction in trout atrial myocytes. In contrast to mammals, cell shortening continued, increasing at membrane potentials above 0 mV in trout atrial myocytes. Furthermore, 5 μM nifedipine abolished L-type Ca2+current ( I Ca) but only reduced cell shortening and the Ca2+ carried by the tail current to 66 ± 5 and 67 ± 6% of the control value. Lowering of the pipette Na+ concentration from 16 to 10 or 0 mM reduced Ca2+ extrusion from the cell from 2.5 ± 0.2 to 1.0 ± 0.2 and 0.5 ± 0.06 amol/pF. With 20 μM exchanger inhibitory peptide (XIP) in the patch pipette Ca2+extrusion 20 min after patch break was 39 ± 8% of its initial value. With 16, 10, and 0 mM Na+ in the pipette, the sarcoplasmic reticulum (SR) Ca2+ content was 47 ± 4, 29 ± 6, and 10 ± 3 amol/pF, respectively. Removal of Na+ from or inclusion of 20 μM XIP in the pipette gradually eliminated the SR Ca2+ content. Whereas I Ca was the same at −10 or +10 mV, Ca2+ extrusion from the cell and the SR Ca2+content at −10 mV were 65 ± 7 and 80 ± 4% of that at +10 mV. The relative amount of Ca2+ extruded by the NCX (about 55%) and taken up by the SR (about 45%) was, however, similar with depolarizations to −10 and +10 mV. We conclude that modulation of the NCX activity critically determines Ca2+ entry and cell shortening in trout atrial myocytes. This is due to both an alteration of the transsarcolemmal Ca2+ transport and a modulation of the SR Ca2+ content.

Permeation and gating properties of a cloned renal K+ channel

1995 ◽  
Vol 268 (2) ◽  
pp. C389-C401 ◽  
Author(s):  
S. Chepilko ◽  
H. Zhou ◽  
H. Sackin ◽  
L. G. Palmer
Keyword(s):  
Single Channel ◽  
Reversal Potential ◽  
Cation Binding ◽  
K Channel ◽  
Patch Clamp Technique ◽  
Open Probability ◽  
Membrane Hyperpolarization ◽  
Channel Current ◽  
Inward Currents ◽  
Kinetics Of

The renal K+ channel (ROMK2) was expressed in Xenopus oocytes, and the patch-clamp technique was used to assess its conducting and gating properties. In cell-attached patches with 110 mM K+ in the bath and pipette, the reversal potential was near zero and the inward conductance (36 pS) was larger than the outward conductance (17 pS). In excised inside-out patches the channels showed rectification in the presence of 5 mM Mg2+ on the cytoplasmic side but not in Mg(2+)-free solution. Inward currents were also observed when K+ was replaced in the pipette by Rb+, NH4+, or thallium (Tl+). The reversal potentials under these conditions yielded a selectivity sequence of Tl+ > K+ > Rb+ > NH4+. On the other hand, the slope conductances for inward current gave a selectivity sequence of K+ = NH4+ > Tl+ > Rb+. The differences in the two sequences can be explained by the presence of cation binding sites within the channel, which interact with Rb+ and Tl+ more strongly and with NH4+ less strongly than with K+. Two other ions, Ba2+ and Cs+, blocked the channel from the outside. The effect of Ba2+ (1 mM) was to reduce the open probability of the channels, whereas Cs+ (10 mM) reduced the apparent single-channel current. The effects of both blockers are enhanced by membrane hyperpolarization. The kinetics of the channel were also studied in cell-attached patches. With K+ in the pipette the distribution of open times could be described by a single exponential (tau 0 = 25 ms), whereas two exponentials (tau 1 = 1 ms, tau 2 = 30 ms) were required to describe the closed-time distribution. Hyperpolarization of the oocyte membrane decreased the open probability and tau 0, and increased tau 1, tau 2, and the number of long closures. The presence of Tl+ in the pipette significantly altered the kinetics, reducing tau 0 and eliminating the long-lived closures. These results suggest that the gating of the channel may depend on the nature of the ion in the pore.

scholarly journals A Trek-Like K+ Channel Current Inhibited by Norepinephrine in Rat Atrial Myocytes

2014 ◽  
Vol 106 (2) ◽  
pp. 120a
Author(s):  
Richard C. Bond ◽  
Stephanie C. Choisy ◽  
Simon M. Bryant ◽  
Jules C. Hancox ◽  
Andrew F. James
Keyword(s):  
K Channel ◽  
Atrial Myocytes ◽  
Channel Current

Abstract 17193: Inhibiting Late Sodium Current Attenuates Isoproterenol-Induced Transient Inward Current and Delayed Afterdepolarizations in Ventricular Myocytes

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Yejia Song ◽  
Nesrine El-Bizri ◽  
Sridharan Rajamani ◽  
Luiz Belardinelli
Keyword(s):  
Ventricular Myocytes ◽  
Sodium Current ◽  
Selective Inhibition ◽  
Adrenergic Stimulation ◽  
Patch Clamp Technique ◽  
Cardiac Tissues ◽  
Inward Current ◽  
Transient Inward Current ◽  
Whole Cell Patch Clamp ◽  
Series Of Experiments

Introduction: The β-adrenergic agonist isoproterenol (ISO) is known to induce the arrhythmogenic transient inward current (I Ti ) and delayed afterdepolarization (DAD) via a stimulation of L-type Ca 2+ current. Recent studies found that ISO-induced DADs in cardiac tissues are inhibited by GS967, a selective blocker of the late Na + current (I NaL ). Thus, we hypothesize that I NaL contributes to the actions of ISO, and selective inhibition of this current will reduce ISO-induced I Ti and DADs. Methods: Transmembrane currents and action potentials of rabbit and guinea pig (GP) ventricular myocytes were recorded using the whole-cell patch-clamp technique. ISO (0.1 μM), GS967 (1 μM) and the Na + channel blocker tetrodotoxin (TTX, 3 μM) were used in the experiments. Results: In rabbit myocytes, application of ISO caused an increase in the amplitude of I NaL from -0.10±0.03 to -0.32±0.04 pA/pF (n = 17, p < 0.05). The ISO-stimulated I NaL was inhibited by GS967 and TTX. In one series of experiments, ISO increased the I NaL from -0.14±0.04 to -0.35±0.06 pA/pF, and GS967 applied in the presence of ISO reduced the current to -0.14±0.03 pA/pF (n = 9, p < 0.05). In another series of experiments, the amplitude of I NaL was increased by ISO from -0.17±0.08 to -0.41±0.09 pA/pF, and was decreased to -0.09±0.08 pA/pF when TTX was applied with ISO (n = 5, p < 0.05). Application of ISO also induced I Ti and DADs. GS967 applied in the presence of ISO inhibited the amplitude of I Ti by 52±6%, from -1.79±0.30 to -0.87±0.16 pA/pF (n = 8, p < 0.05). Consistent with the inhibition of I Ti , GS967 suppressed the amplitude of ISO-induced DADs by 56±12%, from 6.54±1.59 to 3.22±1.27 mV (n = 5, p < 0.05). Similarly, in GP myocytes ISO-induced I Ti and DADs were decreased by GS967 from -1.14±0.21 to -0.73±0.16 pA/pF (n = 7, p < 0.05) and from 7.16±0.59 to 4.67±0.24 mV (n = 5, p < 0.05), respectively. Conclusions: An increased I NaL is likely to contribute to the proarrhythmic effects of ISO in cardiac myocytes. GS967 significantly attenuated ISO-induced I NaL , I Ti and DADs, suggesting that inhibiting this current could be an effective strategy to antagonize the arrhythmogenic actions of β-adrenergic stimulation.

Atrial pressure, distension, and pacing frequency in ANP secretion in isolated perfused rabbit atria

1991 ◽  
Vol 260 (1) ◽  
pp. R39-R46 ◽  
Author(s):  
K. W. Cho ◽  
K. H. Seul ◽  
S. H. Kim ◽  
K. M. Seul ◽  
G. Y. Koh
Keyword(s):  
Electrical Stimulation ◽  
Atrial Natriuretic Peptide ◽  
Natriuretic Peptide ◽  
Atrial Pressure ◽  
Atrial Pacing ◽  
Atrial Myocytes ◽  
Volume Changes ◽  
Series Of Experiments ◽  
The Relationship ◽  
Rabbit Atria

It has been suggested in this laboratory that the principal stimulus for the secretion of atrial natriuretic peptide (ANP) is the reduction of atrial distension and that the secretion of ANP is dependent on both atrial reduction volume and reduction frequency. To investigate the relationship among the changes in atrial pressure, distension, pacing frequency, and ANP secretion, we performed a series of experiments in the isolated perfused rabbit atria. Increase in atrial pressure without changes in transmural pressure and thus without volume changes did not raise immunoreactive ANP (irANP) secretion. Atrial distension without changes in intracavitary atrial pressure increased irANP secretion with the reduction. Electrical stimulation with atrial distension resulted in an increase in irANP secretion in proportion to pacing frequency. Incremental response of irANP secretion to electrical stimulation was accentuated by increasing atrial distension. Neither atrial pacing without distension nor distension without pacing raised irANP secretion. These results suggest that the direct and principal stimulus for irANP secretion in response to atrial pacing and distension is the length shortening of atrial myocytes and that the incremental response of irANP secretion to increasing pacing frequency is the result of an increase in frequency of the length shortening of atrial myocytes.

Inhibition of L-type Ca2+ current by C-type natriuretic peptide in bullfrog atrial myocytes: an NPR-C-mediated effect

2003 ◽  
Vol 285 (6) ◽  
pp. H2454-H2462 ◽  
Author(s):  
R. A. Rose ◽  
A. E. Lomax ◽  
W. R. Giles
Keyword(s):  
Adenylyl Cyclase ◽  
Voltage Clamp ◽  
Natriuretic Peptide ◽  
Guanylyl Cyclase ◽  
Inhibitory Effect ◽  
Inward Rectifier ◽  
Atrial Myocytes ◽  
Adenylyl Cyclase Activity ◽  
Selective Agonist ◽  
Electrophysiological Effects

Single atrial myocytes were isolated from the bullfrog heart and studied under current and voltage clamp conditions to determine the electrophysiological effects of the C-type natriuretic peptide (CNP). CNP (10–8 M) significantly shortened the action potential and reduced its peak amplitude after the application of isoproteronol (10–7 M). In voltage clamp studies, CNP inhibited isoproteronol-stimulated L-type Ca2+ current ( ICa) without any significant effect on the inward rectifier K+ current. The effects of cANF (10–8 M), a selective agonist of the natriuretic peptide C receptor (NPR-C), were very similar to those of CNP. Moreover, HS-142-1, an antagonist of the guanylyl cyclase-linked NPR-A and NPR-B receptors did not alter the inhibitory effect of CNP on ICa. Inclusion of cAMP in the recording pipette to stimulate ICa at a point downstream from adenylyl cyclase increased ICa, but this effect was not inhibited by cANF. These results provide the first demonstration that CNP can inhibit ICa after binding to NPR-C, and suggest that this inhibition involves a decrease in adenylyl cyclase activity, which leads to reduced intracellular levels of cAMP.

The function of the sarcoplasmic reticulum is not inhibited by low temperatures in trout atrial myocytes

2001 ◽  
Vol 281 (6) ◽  
pp. R1902-R1906 ◽  
Author(s):  
Leif Hove-Madsen ◽  
Anna Llach ◽  
Lluis Tort
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Low Temperatures ◽  
Experimental Temperature ◽  
Effect Of Temperature ◽  
Fast Time ◽  
Atrial Myocytes ◽  
Patch Clamp Technique ◽  
Two Temperatures ◽  
Perforated Patch Clamp ◽  
The Relationship

The effect of temperature on sarcoplasmic reticulum (SR) Ca2+ uptake and release was measured in trout atrial myocytes using the perforated patch-clamp technique. Depolarization of the myocyte for 10 s to different membrane potentials ( V m) induced SR Ca2+ uptake. The relationship between V m and SR Ca2+ uptake was not significantly changed by lowering the experimental temperature from 21 to 7°C, and the relationship between total cytosolic Ca2+and SR Ca2+ uptake was similar at the two temperatures with a pooled Vmax = 66 amol/pF and K 0.5 = 4 amol/pF. Quantification of the Ca2+ release from the SR elicited by 10-ms depolarizations to different V m showed an increasing SR Ca2+ release at more positive V mbetween −50 and +10 mV, whereas SR Ca2+ release stagnated between +10 and +50 mV. Lowering of the temperature did not affect this relationship significantly, giving an SR Ca2+ release of 1.71 and 1.54 amol/pF at 21 and 7°C, respectively. Furthermore, clearance of the SR Ca2+ content slowed down inactivation of the L-type Ca2+ current at both temperatures (the fast time constant increased significantly from 10.4 ± 1.9 to 15.0 ± 2.0 ms at 21°C and from 38 ± 15 to 73 ± 24 ms at 7°C). Thus the SR has the capacity to remove the entire Ca2+ transient at physiologically relevant stimulation frequencies at both 21 and 7°C, although it is estimated that ∼40% of the total Ca2+ transient is liberated from and reuptaken by the SR with continuous stimulation at 0.5 Hz independently of the experimental temperature.

Differential Modulation of the Cardiac Adenosine Triphosphate-sensitive Potassium Channel by Isoflurane and Halothane

2002 ◽  
Vol 97 (1) ◽  
pp. 50-56 ◽  
Author(s):  
Wai-Meng Kwok ◽  
Anne T. Martinelli ◽  
Kazuhiro Fujimoto ◽  
Akihiro Suzuki ◽  
Anna Stadnicka ◽  
...  
Keyword(s):  
Potassium Channel ◽  
Adenosine Triphosphate ◽  
Volatile Anesthetic ◽  
Volatile Anesthetics ◽  
Dependent Manner ◽  
Patch Clamp Technique ◽  
Channel Activation ◽  
Beneficial Effects ◽  
Channel Current ◽  
Channel Opening

Background The cardiac adenosine triphosphate-sensitive potassium (K(ATP)) channel is activated during pathophysiological episodes such as ischemia and hypoxia and may lead to beneficial effects on cardiac function. Studies of volatile anesthetic interactions with the cardiac K(ATP) channel have been limited. The goal of this study was to investigate the ability of volatile anesthetics halothane and isoflurane to modulate the cardiac sarcolemmal K(ATP) channel. Methods The K(ATP) channel current (I(KATP)) was monitored using the whole cell configuration of the patch clamp technique from single ventricular cardiac myocytes enzymatically isolated from guinea pig hearts. I(KATP) was elicited by extracellular application of the potassium channel openers 2,4-dinitrophenol or pinacidil. Results Volatile anesthetics modulated I(KATP) in an anesthetic-dependent manner. Isoflurane facilitated the opening of the K(ATP) channel. Following initial activation of I(KATP) by 2,4-dinitrophenol, isoflurane at 0.5 and 1.3 mm further increased current amplitude by 40.4 +/- 11.1% and 58.4 +/- 20.6%, respectively. Similar results of isoflurane were obtained when pinacidil was used to activate I(KATP). However, isoflurane alone was unable to elicit K(ATP) channel opening. In contrast, halothane inhibited I(KATP) elicited by 2,4-dinitrophenol by 50.6 +/- 5.8% and 72.1 +/- 11.6% at 0.4 and 1.0 mm, respectively. When I(KATP) was activated by pinacidil, halothane had no significant effect on the current. Conclusions The cardiac sarcolemmal K(ATP) channel is differentially modulated by volatile anesthetics. Isoflurane can facilitate the further opening of the K(ATP) channel following initial channel activation by 2,4-dinitrophenol or pinacidil. The effect of halothane was dependent on the method of channel activation, inhibiting I(KATP) activated by 2,4-dinitrophenol but not by pinacidil.

scholarly journals Inhibition of L-type calcium-channel activity by thapsigargin and 2,5-t-butylhydroquinone, but not by cyclopiazonic acid

1994 ◽  
Vol 302 (1) ◽  
pp. 147-154 ◽  
Author(s):  
E J Nelson ◽  
C C R Li ◽  
R Bangalore ◽  
T Benson ◽  
R S Kass ◽  
...  
Keyword(s):  
Calcium Channels ◽  
Calcium Channel ◽  
Cyclopiazonic Acid ◽  
Calcium Currents ◽  
Pituitary Cells ◽  
Channel Activity ◽  
Patch Clamp Technique ◽  
Channel Current ◽  
Low Concentrations ◽  
Ic50 Values

Thapsigargin (TG), 2,5-t-butylhydroquinone (tBHQ) and cyclopiazonic acid (CPA) all inhibit the initial Ca(2+)-response to thyrotropin-releasing hormone (TRH) by depleting intracellular Ca2+ pools sensitive to inositol 1,4,5-trisphosphate (IP3). Treatment of GH3 pituitary cells for 30 min with 5 nM TG, 500 nM tBHQ or 50 nM CPA completely eliminated the TRH-induced spike in intracellular free Ca2+ ([Ca2+]i). Higher concentrations of TG and tBHQ, but not CPA, were also found to inhibit strongly the activity of L-type calcium channels, as measured by the increase in [Ca2+]i or 45Ca2+ influx stimulated by depolarization. TG and tBHQ blocked high-K(+)-stimulated 45Ca2+ uptake, with IC50 values of 10 and 1 microM respectively. Maximal inhibition of L-channel activity was achieved 15-30 min after drug addition. Inhibition by tBHQ was reversible, whereas inhibition by TG was not. TG and CPA did not affect spontaneous [Ca2+]i oscillations when tested at concentrations adequate to deplete the IP3-sensitive Ca2+ pool. However, 20 microM TG and 10 microM tBHQ blocked [Ca2+]i oscillations completely. The effect of drugs on calcium currents was measured directly by using the patch-clamp technique. When added to the external bath, 10 microM CPA caused a sustained increase in the calcium-channel current amplitude over 8 min, 10 microM tBHQ caused a progressive inhibition, and 10 microM TG caused an enhancement followed by a sustained block of the calcium current over 8 min. In summary, CPA depletes IP3-sensitive Ca2+ stores and does not inhibit voltage-operated calcium channels. At sufficiently low concentrations, TG depletes IP3-sensitive stores without inhibiting L-channel activity, but, for tBHQ, inhibition of calcium channels occurs at concentrations close to those needed to block agonist mobilization of intracellular Ca2+.

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