Ionic Currents Underlying Fast Action Potentials in the Obliquely Striated Muscle Cells of the Octopus Arm

2002 ◽  
Vol 88 (6) ◽  
pp. 3386-3397 ◽  
Author(s):  
Dan Rokni ◽  
Binyamin Hochner
Keyword(s):  
Time Constant ◽  
Action Potentials ◽  
Striated Muscle ◽  
Muscle Cells ◽  
Ionic Currents ◽  
Delayed Rectifier ◽  
Inactivation Kinetics ◽  
Activation Kinetics ◽  
Voltage Dependent ◽  
Neuromuscular Systems

The octopus arm provides a unique model for neuromuscular systems of flexible appendages. We previously reported the electrical compactness of the arm muscle cells and their rich excitable properties ranging from fast oscillations to overshooting action potentials. Here we characterize the voltage-activated ionic currents in the muscle cell membrane. We found three depolarization-activated ionic currents: 1) a high-voltage-activated L-type Ca2+ current, which began activating at approximately −35 mV, was eliminated when Ca2+ was substituted by Mg2+, was blocked by nifedipine, and showed Ca2+-dependent inactivation. This current had very rapid activation kinetics (peaked within milliseconds) and slow inactivation kinetics (τ in the order of 50 ms). 2) A delayed rectifier K+ current that was totally blocked by 10 mM TEA and partially blocked by 10 mM 4-aminopyridine (4AP). This current exhibited relatively slow activation kinetics (τ in the order of 15 ms) and inactivated only partially with a time constant of ∼150 ms. And 3) a transient A-type K+ current that was totally blocked by 10 mM 4AP and was partially blocked by 10 mM TEA. This current exhibited very fast activation kinetics (peaked within milliseconds) and inactivated with a time constant in the order of 60 ms. Inactivation of the A-type current was almost complete at −40 mV. No voltage-dependent Na+ current was found in these cells. The octopus arm muscle cells generate fast (∼3 ms) overshooting spikes in physiological conditions that are carried by a slowly inactivating L-type Ca2+ current.

Effects of melatonin on ionic currents in cultured ocular tissues

1999 ◽  
Vol 276 (4) ◽  
pp. C923-C929 ◽  
Author(s):  
Adam Rich ◽  
Gianrico Farrugia ◽  
James L. Rae
Keyword(s):  
Epithelial Cells ◽  
Trabecular Meshwork ◽  
Cell Voltage ◽  
Ionic Currents ◽  
Lens Epithelial Cells ◽  
Delayed Rectifier ◽  
Inactivation Kinetics ◽  
Epithelial Cell Line ◽  
Voltage Dependent ◽  
Mouse Lens

The effects of melatonin on ionic conductances in a cultured mouse lens epithelial cell line (α-TN4) and in cultured human trabecular meshwork (HTM) cells were measured using the amphotericin perforated patch whole cell voltage-clamp technique. Melatonin stimulated a voltage-dependent Na+-selective current in lens epithelial cells and trabecular meshwork cells. The effects of melatonin were observed at 50 pM and were maximal at 100 μM. Melatonin enhanced activation and inactivation kinetics, but no change was observed in the voltage dependence of activation. The results are consistent with an increase in the total number of ion channels available for activation by membrane depolarization. Melatonin was also found to stimulate a K+-selective current at high doses (1 mM). Melatonin did not affect the inwardly rectifying K+ current or the delayed rectifier type K+ current that has been described in cultured mouse lens epithelial cells. The results show that melatonin specifically stimulated the TTX-insensitive voltage-dependent Na+ current by an apparently novel mechanism.

Two Types of Voltage-Gated K+ Currents in Dissociated Heart Ventricular Muscle Cells of the Snail Lymnaea stagnalis

1999 ◽  
Vol 82 (5) ◽  
pp. 2415-2427 ◽  
Author(s):  
M. S. Yeoman ◽  
P. R. Benjamin
Keyword(s):  
Voltage Clamp ◽  
Action Potentials ◽  
Muscle Cells ◽  
Delayed Rectifier ◽  
Voltage Sensitivity ◽  
Ventricular Muscle ◽  
Current Time ◽  
Voltage Gated ◽  
Time To Peak ◽  
Voltage Dependent

We have used a combination of current-clamp and voltage-clamp techniques to characterize the electrophysiological properties of enzymatically dissociated Lymnaea heart ventricle cells. Dissociated ventricular muscle cells had average resting membrane potentials of −55 ± 5 mV. When hyperpolarized to potentials between −70 and −63 mV, ventricle cells were capable of firing repetitive action potentials (8.5 ± 1.2 spikes/min) that failed to overshoot 0 mV. The action potentials were either simple spikes or more complex spike/plateau events. The latter were always accompanied by strong contractions of the muscle cell. The waveform of the action potentials were shown to be dependent on the presence of extracellular Ca2+ and K+ ions. With the use of the single-electrode voltage-clamp technique, two types of voltage-gated K+ currents were identified that could be separated by differences in their voltage sensitivity and time-dependent kinetics. The first current activated between −50 and −40 mV. It was relatively fast to activate (time-to-peak; 13.7 ± 0.7 ms at +40 mV) and inactivated by 53.3 ± 4.9% during a maintained 200-ms depolarization. It was fully available for activation below −80 mV and was completely inactivated by holding potentials more positive than −40 mV. It was completely blocked by 5 mM 4-aminopyridine (4-AP) and by concentrations of tetraethylammonium chloride (TEA) >10 mM. These properties characterize this current as a member of the A-type family of voltage-dependent K+ currents. The second voltage-gated K+ current activated at more depolarized potentials (−30 to −20 mV). It activated slower than the A-type current (time-to-peak; 74.1 ± 3.9 ms at +40 mV) and showed little inactivation (6.2 ± 2.1%) during a maintained 200-ms depolarization. The current was fully available for activation below −80 mV with a proportion of the current still available for activation at potentials as positive as 0 mV. The current was completely blocked by 1–3 mM TEA. These properties characterize this current as a member of the delayed rectifier family of voltage-dependent K+ currents. The slow activation rates and relatively depolarized activation thresholds of the two K+ currents are suggestive that their main role is to contribute to the repolarization phase of the action potential.

Altered Regulation of Potassium and Calcium Channels by GABAB and Adenosine Receptors in Hippocampal Neurons From Mice Lacking Gαo

2000 ◽  
Vol 83 (2) ◽  
pp. 1010-1018 ◽  
Author(s):  
Gabriela J. Greif ◽  
Deborah L. Sodickson ◽  
Bruce P. Bean ◽  
Eva J. Neer ◽  
Ulrike Mende
Keyword(s):  
Time Constant ◽  
Hippocampal Neurons ◽  
Ionic Currents ◽  
Absolute Requirement ◽  
Voltage Dependent ◽  
Hippocampal Ca3 ◽  
Ca3 Neurons ◽  
Inwardly Rectifying ◽  
G Protein Coupled

To examine the role of Go in modulation of ion channels by neurotransmitter receptors, we characterized modulation of ionic currents in hippocampal CA3 neurons from mice lacking both isoforms of Gαo. In CA3 neurons from Gαo −/− mice, 2-chloro-adenosine and the GABAB-receptor agonist baclofen activated inwardly rectifying K+ currents and inhibited voltage-dependent Ca2+ currents just as effectively as in Gαo +/+ littermates. However, the kinetics of transmitter action were dramatically altered in Gαo −/− mice in that recovery on washout of agonist was much slower. For example, recovery from 2-chloro-adenosine inhibition of calcium current was more than fourfold slower in neurons from Gαo −/− mice [time constant of 12.0 ± 0.8 (SE) s] than in neurons from Gαo +/+ mice (time constant of 2.6 ± 0.2 s). Recovery from baclofen effects was affected similarly. In neurons from control mice, effects of both baclofen and 2-chloro-adenosine on Ca2+ currents and K+currents were abolished by brief exposure to external N-ethyl-maleimide (NEM). In neurons lacking Gαo, some inhibition of Ca2+ currents by baclofen remained after NEM treatment, whereas baclofen activation of K+ currents and both effects of 2-chloro-adenosine were abolished. These results show that modulation of Ca2+ and K+ currents by G protein-coupled receptors in hippocampal neurons does not have an absolute requirement for Gαo. However, modulation is changed in the absence of Gαo in having much slower recovery kinetics. A likely possibility is that the very abundant Gαo is normally used but, when absent, can readily be replaced by G proteins with different properties.

Input Resistance, Time Constants, and Spike Initiation

1998 ◽  
Author(s):  
Christof Koch
Keyword(s):  
Membrane Potential ◽  
Time Constant ◽  
Action Potentials ◽  
Input Resistance ◽  
Current Injection ◽  
Membrane Conductances ◽  
Voltage Dependent ◽  
Dc Component ◽  
Definition Of ◽  
A Current

This chapter represents somewhat of a tephnical interlude. Having introduced the reader to both simplified and more complex compartmental single neuron models, we need to revisit terrain with which we are already somewhat familiar. In the following pages we reevaluate two important concepts we defined in the first few chapters: the somatic input resistance and the neuronal time constant. For passive systems, both are simple enough variables: Rin is the change in somatic membrane potential in response to a small sustained current injection divided by the amplitude of the current injection, while τm is the slowest time constant associated with the exponential charging or discharging of the neuronal membrane in response to a current pulse or step. However, because neurons express nonstationary and nonlinear membrane conductances, the measurement and interpretation of these two variables in active structures is not as straightforward as before. Having obtained a more sophisticated understanding of these issues, we will turn toward the question of the existence of a current, voltage, or charge threshold at which a biophysical faithful model of a cell triggers action potentials. We conclude with recent work that suggests how concepts from the subthreshold domain, like the input resistance or the average membrane potential, could be extended to the case in which the cell is discharging a stream of action potentials. This chapter is mainly for the cognoscendi or for those of us that need to make sense of experimental data by comparing therp to theoretical models that usually fail to reflect reality adequately. In Sec. 3.4, we defined Kii (f) for passive cable structures as the voltage change at location i in response to a sinusoidal current injection of frequency f at the same location. Its dc component is also referred to as input resistance or Rin. Three difficulties render this definition of input resistance problematic in real cells: (1) most membranes, in particular at the soma, show voltage-dependent nonlinearities, (2) the associated ionic membrane conductances are time dependent and (3) instrumental aspects, such as the effect of the impedance of the recording electrode on Rin, add uncertainty to the measuring process.

Expression of voltage-dependent K+ channel genes in mesenteric artery smooth muscle cells

1999 ◽  
Vol 277 (5) ◽  
pp. G1055-G1063 ◽  
Author(s):  
Chuanli Xu ◽  
Yanjie Lu ◽  
Guanghua Tang ◽  
Rui Wang
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Mesenteric Artery ◽  
Muscle Cells ◽  
Mesenteric Arteries ◽  
Delayed Rectifier ◽  
Kv Channel ◽  
Channel Proteins ◽  
Voltage Dependent ◽  
Encoding Genes

Molecular basis of native voltage-dependent K+(Kv) channels in smooth muscle cells (SMCs) from rat mesenteric arteries was investigated. The whole cell patch-clamp study revealed that a 4-aminopyridine-sensitive delayed rectifier K+ current ( I K) was the predominant K+ conductance in these cells. A systematic screening of the expression of 18 Kv channel genes using RT-PCR technique showed that six I K-encoding genes (Kv1.2, Kv1.3, Kv1.5, Kv2.1, Kv2.2, and Kv3.2) were expressed in mesenteric artery. Although no transient outward Kv currents ( I A) were recorded in the studied SMCs, transcripts of multiple I A-encoding genes, including Kv1.4, Kv3.3, Kv3.4, Kv4.1, Kv4.2, and Kv4.3 as well as I A-facilitating Kv β-subunits (Kvβ1, Kvβ2, and Kvβ3), were detected in mesenteric arteries. Western blot analysis demonstrated that four I K-related Kv channel proteins (Kv1.2, Kv1.3, Kv1.5, and Kv2.1) were detected in mesenteric artery tissues. The presence of Kv1.2, Kv1.3, Kv1.5, and Kv2.1 channel proteins in isolated SMCs was further confirmed by immunocytochemistry study. Our results suggest that the native I K in rat mesenteric artery SMCs might be generated by heteromultimerization of Kv genes.

scholarly journals High purity human-induced pluripotent stem cell-derived cardiomyocytes: electrophysiological properties of action potentials and ionic currents

2011 ◽  
Vol 301 (5) ◽  
pp. H2006-H2017 ◽  
Author(s):  
Junyi Ma ◽  
Liang Guo ◽  
Steve J. Fiene ◽  
Blake D. Anson ◽  
James A. Thomson ◽  
...  
Keyword(s):  
Cardiac Myocytes ◽  
High Purity ◽  
Action Potentials ◽  
Ionic Currents ◽  
Delayed Rectifier ◽  
Cardiovascular Research ◽  
Peak Voltage ◽  
Electrophysiological Properties ◽  
Induced Pluripotent ◽  
Multiple Drugs

Human-induced pluripotent stem cells (hiPSCs) can differentiate into functional cardiomyocytes; however, the electrophysiological properties of hiPSC-derived cardiomyocytes have yet to be fully characterized. We performed detailed electrophysiological characterization of highly pure hiPSC-derived cardiomyocytes. Action potentials (APs) were recorded from spontaneously beating cardiomyocytes using a perforated patch method and had atrial-, nodal-, and ventricular-like properties. Ventricular-like APs were more common and had maximum diastolic potentials close to those of human cardiac myocytes, AP durations were within the range of the normal human electrocardiographic QT interval, and APs showed expected sensitivity to multiple drugs (tetrodotoxin, nifedipine, and E4031). Early afterdepolarizations (EADs) were induced with E4031 and were bradycardia dependent, and EAD peak voltage varied inversely with the EAD take-off potential. Gating properties of seven ionic currents were studied including sodium ( INa), L-type calcium ( ICa), hyperpolarization-activated pacemaker ( If), transient outward potassium ( Ito), inward rectifier potassium ( IK1), and the rapidly and slowly activating components of delayed rectifier potassium ( IKr and IKs, respectively) current. The high purity and large cell numbers also enabled automated patch-clamp analysis. We conclude that these hiPSC-derived cardiomyocytes have ionic currents and channel gating properties underlying their APs and EADs that are quantitatively similar to those reported for human cardiac myocytes. These hiPSC-derived cardiomyocytes have the added advantage that they can be used in high-throughput assays, and they have the potential to impact multiple areas of cardiovascular research and therapeutic applications.

Effect of sodium nitroprusside on Ca2+ currents in opossum esophageal circular muscle cells

1994 ◽  
Vol 266 (6) ◽  
pp. G1036-G1042 ◽  
Author(s):  
H. I. Akbarali ◽  
R. K. Goyal
Keyword(s):  
Sodium Nitroprusside ◽  
Voltage Clamp ◽  
Action Potentials ◽  
Circular Muscle ◽  
Outward Current ◽  
Muscle Cells ◽  
Ionic Currents ◽  
Inhibitory Effect ◽  
High K ◽  
Direct Inhibitory Effect

The effects of sodium nitroprusside (SNP) on ionic currents in single opossum esophageal circular muscle cells were examined. In voltage clamp, Ca2+ currents were studied after K+ currents were blocked with Cs+ in the patch pipette. The threshold for inward Ca2+ currents was -30 mV with peak current between 0 and +10 mV from holding potentials of -90 mV. The Ca2+ currents had both transient and sustained phases. The transient phase was partially resistant to nifedipine (1 microM). SNP (100 microM) reversibly decreased both the transient and sustained phases of the Ca2+ currents by approximately 20%. In cells dialyzed with high-K+ solutions, voltage-clamp recordings demonstrated the presence of an inward current followed by an outward current at potentials positive to -30 mV. SNP under these conditions resulted in a decrease in the Ca2+ current and decreased the outward current during test depolarizations. Action potentials were evoked during current-clamp recordings that consisted of multiple spikes, depending on the stimulus strength. The threshold for spike generation was close to -30 mV and was blocked by Cd2+, suggesting that the upstroke of the action potential was dependent on Ca2+ influx. SNP significantly attenuated action potentials and produced a small hyperpolarization (5-7 mV). These results suggest that SNP has a direct inhibitory effect on Ca2+ currents and thereby decreases evoked action potentials, and that SNP hyperpolarization is not due to Ca(2+)-activated K+ channels.

Characterization of macroscopic outward currents of canine colonic myocytes

1989 ◽  
Vol 257 (3) ◽  
pp. C461-C469 ◽  
Author(s):  
W. C. Cole ◽  
K. M. Sanders
Keyword(s):  
Outward Current ◽  
Cell Voltage ◽  
Muscle Cells ◽  
Time Dependent ◽  
Delayed Rectifier ◽  
Delayed Rectifier Current ◽  
Outward Currents ◽  
Voltage Dependent ◽  
K Current ◽  
Time Courses

Outward currents of colonic smooth muscle cells were characterized by the whole cell voltage-clamp method. Four components of outward current were identified: a time-independent and three time-dependent components. The time-dependent current showed strong outward rectification positive to -25 mV and was blocked by tetraethylammonium. The time-dependent components were separated on the basis of their time courses, voltage dependence, and pharmacological sensitivities. They are as follows. 1) A Ca2+-activated K current sensitive to external Ca2+ and Ca2+ influx was blocked by ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (0.1 X 10(-3) M) and nifedipine (1 X 10(-6) and was increased by elevated Ca2+ (8 X 10(-6) M) and BAY K 8644 (1 X 10(-6) M). 2) A "delayed rectifier" current was observed that decayed slowly with time and showed no voltage-dependent inactivation. 3) Spontaneous transient outward currents that were blocked by ryanodine (2 X 10(-6) M) were also recorded. The possible contributions of these currents to the electrical activity of colonic muscle cells in situ are discussed. Ca2+-activated K current may contribute a significant conductance to the repolarizing phase of electrical slow waves.

Ion Channel Compartments in Photoreceptors: Evidence From Salamander Rods With Intact and Ablated Terminals

2007 ◽  
Vol 98 (1) ◽  
pp. 86-95 ◽  
Author(s):  
Peter R. MacLeish ◽  
Colin A. Nurse
Keyword(s):  
Ion Channels ◽  
Imaging Techniques ◽  
Cell Voltage ◽  
Ionic Currents ◽  
Simultaneous Recording ◽  
Delayed Rectifier ◽  
Sensory Cells ◽  
Before And After ◽  
Voltage Dependent ◽  
Highly Correlated

Vertebrate photoreceptors are highly polarized sensory cells in which several different ionic currents have been characterized. In the present study we used whole cell voltage-clamp and optical imaging techniques, the former combined with microsurgical manipulations, and simultaneous recording of membrane current and intracellular calcium signals to investigate the spatial distribution of ion channels within isolated salamander rods. In recordings from intact rods with visible terminals, evidence for five previously identified ionic currents was obtained. These include two Ca2+-dependent, i.e., a Ca2+-dependent chloride current [ ICl(Ca)] and a large-conductance Ca2+- and voltage-dependent K+ or BK current [ IK(Ca)], and three voltage-dependent currents, i.e., a delayed-rectifier type current [ IK(V)], a hyperpolarization-activated cation current ( Ih), and a dihydropyridine-sensitive L-type calcium current ( ICa). Of these, ICl(Ca) was highly correlated with the presence of a terminal; rods with visible terminals expressed ICl(Ca) without exception ( n = 125), whereas approximately 71% of rods (40/56) without visible terminals lacked ICl(Ca). More significantly, ICl(Ca) was absent from all rods ( n = 33) that had their terminals ablated, and recordings from the same cell before and after terminal ablation led, in all cases ( n =10), to the loss of ICl(Ca). In contrast, IK(Ca), IK(V), and Ih remained largely intact after terminal ablation, suggesting that they arose principally from ion channels located in the soma and/or inner segment. The outward IK(Ca) in terminal-ablated rods was reversibly suppressed on “puffing” a Ca2+-free extracellular solution over the soma and was appreciably enhanced by the L-type Ca2+ channel agonist, Bay K 8644 (0.1–2 μM). These data indicate that rod photoreceptors possess discrete targeting mechanisms that preferentially sort ion channels mediating ICl(Ca) to the terminal.

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