Electrotonic suppression of early afterdepolarizations in isolated rabbit Purkinje myocytes

2000 ◽  
Vol 279 (1) ◽  
pp. H250-H259 ◽  
Author(s):  
Delilah J. Huelsing ◽  
Kenneth W. Spitzer ◽  
Andrew E. Pollard
Keyword(s):  
Ventricular Myocytes ◽  
Single Cells ◽  
Late Phase ◽  
Input Resistance ◽  
Membrane Resistance ◽  
Early Afterdepolarizations ◽  
Model Cell ◽  
Triggered Arrhythmias ◽  
Electrotonic Interactions ◽  
Coupling Current

Many studies suggest that early afterdepolarizations (EADs) arising from Purkinje fibers initiate triggered arrhythmias under pathological conditions. However, electrotonic interactions between Purkinje and ventricular myocytes may either facilitate or suppress EAD formation at the Purkinje-ventricular interface. To determine conditions that facilitated or suppressed EADs during Purkinje-ventricular interactions, we coupled single Purkinje myocytes and aggregates isolated from rabbit hearts to a passive model cell via an electronic circuit with junctional resistance ( R j). The model cell had input resistance ( R m,v) of 50 MΩ, capacitance of 39 pF, and a variable rest potential ( V rest,v). EADs were induced in Purkinje myocytes during superfusion with 1 μM isoproterenol. Coupling at high R j to normally polarized V rest,v established a repolarizing coupling current during all phases of the Purkinje action potential. This coupling current preferentially suppressed EADs in single cells with mean membrane resistance ( R m,p) of 297 MΩ, whereas EAD suppression in larger aggregates with mean R m,p of 80 MΩ required larger coupling currents. In contrast, coupling to elevated V rest,v established a depolarizing coupling current during late phase 2, phase 3, and phase 4 that facilitated EAD formation and induced spontaneous activity in single Purkinje myocytes and aggregates. These results have important implications for arrhythmogenesis in the infarcted heart when reduction of the ventricular mass due to scarring alters the R m,p-to- R m,v ratio and in the ischemic heart when injury currents are established during coupling between polarized Purkinje myocytes and depolarized ventricular myocytes.

Membrane resistance increases when automaticity develops in explanted rat heart cells

1990 ◽  
Vol 258 (1) ◽  
pp. H145-H152 ◽  
Author(s):  
O. F. Schanne ◽  
M. Lefloch ◽  
B. Fermini ◽  
E. Ruiz-Petrich
Keyword(s):  
Spontaneous Activity ◽  
Action Potentials ◽  
Ventricular Myocytes ◽  
Input Resistance ◽  
Membrane Resistance ◽  
Resting Potential ◽  
Heart Cells ◽  
Membrane Capacity ◽  
Rat Heart Cells ◽  
Specific Membrane

We compared the passive electrical properties of isolated ventricular myocytes (resting potential -65 mV, fast action potentials, and no spontaneous activity) with those of 2- to 7-day-old cultured ventricle cells from neonatal rats (resting potential -50 mV, slow action potentials, and presence of spontaneous activity). In myocytes the specific membrane capacity was 0.99 microF/cm2, and the specific membrane resistance increased from 2.46 k omega.cm2 at -65 mV to 7.30 k omega.cm2 at -30 mV. In clusters, the current-voltage relationships measured under current-clamp conditions showed anomalous rectification and the input resistance decreased from 1.05 to 0.48 M omega when external K+ concentration was increased from 6 to 100 mM. Using the model of a finite disk we determined the specific membrane resistance (12.9 k omega.cm2), the effective membrane capacity (17.8 microF/cm2), and the lumped resistivity of the disk interior (1,964 omega.cm). We conclude that 1) the voltage dependence of the specific membrane resistance cannot completely explain the membrane resistance increase that accompanies the appearance of spontaneous activity; 2) a decrease of the inwardly rectifying conductance (gk1) is mainly responsible for the increase in the specific membrane resistance and depolarization; and 3) approximately 41% of the inward-rectifying channels are electrically silent when spontaneous activity develops in explanted ventricle cells.

Intracellular calcium activates a chloride current in canine ventricular myocytes

1994 ◽  
Vol 267 (5) ◽  
pp. H1984-H1995 ◽  
Author(s):  
A. C. Zygmunt
Keyword(s):  
Potassium Current ◽  
Ventricular Myocytes ◽  
Single Cells ◽  
Reversal Potential ◽  
Outward Current ◽  
Chloride Current ◽  
Disulfonic Acid ◽  
Transient Outward Current ◽  
Triggered Arrhythmias ◽  
Inward Currents

The contribution of chloride and potassium to the 4-aminopyridine (4-AP)-resistant transient outward current was investigated in dog cardiac myocytes. Whole cell currents were recorded at 37 degrees C in single cells dissociated from epicardial and midmyocardial regions of the canine ventricle. Sodium-calcium exchange current and voltage-dependent transient outward potassium current (IA) were blocked in sodium-free solutions containing 2 mM 4-AP; sodium channels were inactivated by the -50-mV holding potential. When patch pipettes contained 0.4–0.8 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, voltage-clamp steps over the range -20 to +50 mV activated an inward calcium current (ICa) and a Ca(2+)-activated chloride current [ICl(Ca)]. ICl(Ca) was blocked by 200 microM 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, 1 mM 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS), or reduction of external chloride. Independent of the presence of potassium, the reversal potential of the SITS-sensitive current varied with extracellular chloride, as predicted for a chloride-selective conductance. The bell-shaped current-voltage relation of ICl(Ca) has a threshold of -20 mV and a peak at +40 mV. No evidence could be found for a Ca(2+)-activated potassium current or a Ca(2+)-activated nonspecific cation current under these conditions. ICl(Ca) contributed to oscillatory inward currents at diastolic potentials in cells superfused by isoproterenol and high Ca2+, suggesting a role for this current in triggered arrhythmias associated with delayed afterdepolarizations. In the normal heart, ICl(Ca) is likely to contribute to rate- and rhythm-dependent repolarization of the cardiac action potential.

Voltage clamping single cells in intact Malpighian tubules of mosquitoes

2000 ◽  
Vol 279 (4) ◽  
pp. F747-F754 ◽  
Author(s):  
R. Masia ◽  
D. Aneshansley ◽  
W. Nagel ◽  
R. J. Nachman ◽  
K. W. Beyenbach
Keyword(s):  
Apical Membrane ◽  
Single Cells ◽  
Basolateral Membrane ◽  
Malpighian Tubule ◽  
Input Resistance ◽  
Membrane Resistance ◽  
Malpighian Tubules ◽  
Shunt Resistance ◽  
Transport Pathways ◽  
Principal Cells

Principal cells of the Malpighian tubule of the yellow fever mosquito were studied with the methods of two-electrode voltage clamp (TEVC). Intracellular voltage ( V pc) was −86.7 mV, and input resistance ( R pc) was 388.5 kΩ ( n = 49 cells). In six cells, Ba2+ (15 mM) had negligible effects on V pc, but it increased R pc from 325.3 to 684.5 kΩ ( P< 0.001). In the presence of Ba2+, leucokinin-VIII (1 μM) increased V pc to −101.8 mV ( P < 0.001) and reduced R pc to 340.2 kΩ ( P < 0.002). Circuit analysis yields the following: basolateral membrane resistance, 652.0 kΩ; apical membrane resistance, 340.2 kΩ; shunt resistance ( R sh), 344.3 kΩ; transcellular resistance, 992.2 kΩ. The fractional resistance of the apical membrane (0.35) and the ratio of transcellular resistance and R sh (3.53) agree closely with values obtained by cable analysis in isolated perfused tubules and confirm the usefulness of TEVC methods in single principal cells of the intact Malpighian tubule. Dinitrophenol (0.1 mM) reversibly depolarized V pc from −94.3 to −10.7 mV ( P< 0.001) and reversibly increased R pc from 412 to 2,879 kΩ ( P < 0.001), effects that were duplicated by cyanide (0.3 mM). Significant effects of metabolic inhibition on voltage and resistance suggest a role of ATP in electrogenesis and the maintenance of conductive transport pathways.

Beat-to-beat repolarization variability in ventricular myocytes and its suppression by electrical coupling

2000 ◽  
Vol 278 (3) ◽  
pp. H677-H687 ◽  
Author(s):  
Massimiliano Zaniboni ◽  
Andrew E. Pollard ◽  
Lin Yang ◽  
Kenneth W. Spitzer
Keyword(s):  
Ventricular Myocytes ◽  
Single Cells ◽  
Electrical Coupling ◽  
Delayed Rectifier ◽  
Delayed Rectifier Current ◽  
Constant Rate ◽  
Electrotonic Interactions ◽  
Single Ventricular Myocytes ◽  
The Mean ◽  
Junctional Resistance

Single ventricular myocytes paced at a constant rate and held at a constant temperature exhibit beat-to-beat variations in action potential duration (APD). In this study we sought to quantify this variability, assess its mechanism, and determine its responsiveness to electrotonic interactions with another myocyte. Interbeat APD90 (90% repolarization) of single cells was normally distributed. We thus quantified APD90 variability as the coefficient of variability, CV = (SD/mean APD90) × 100. The mean ± SD of the CV in normal solution was 2.3 ± 0.9 (132 cells). Extracellular TTX (13 μM) and intracellular EGTA (14 mM) both significantly reduced the CV by 44 and 26%, respectively. When applied in combination the CV fell by 54%. In contrast, inhibition of the rapid delayed rectifier current with L-691,121 (100 nM) increased the CV by 300%. The CV was also significantly reduced by 35% when two normal myocytes were electrically connected with a junctional resistance ( R j) of 100 MΩ. Electrical coupling ( R j = 100 MΩ) of a normal myocyte to one producing early afterdepolarization (EAD) completely blocked EAD formation. These results indicate that beat-to-beat APD variability is likely mediated by stochastic behavior of ion channels and that electrotonic interactions act to limit temporal dispersion of refractoriness, a major contributor to arrhythmogenesis.

scholarly journals Enhanced activity of ventricular Na+-HCO3− cotransport in pressure overload hypertrophy

2007 ◽  
Vol 293 (2) ◽  
pp. H1254-H1264 ◽  
Author(s):  
Taku Yamamoto ◽  
Takeshi Shirayama ◽  
Tomohiko Sakatani ◽  
Tomosaburo Takahashi ◽  
Hideo Tanaka ◽  
...  
Keyword(s):  
Ventricular Myocytes ◽  
Single Cells ◽  
Ischemia Reperfusion ◽  
Pressure Overload ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Aortic Constriction ◽  
Ph Indicator ◽  
Adult Rat ◽  
Acid Extrusion

The Na+-HCO3− cotransporter (NBC) plays a key role in intracellular pH (pHi) regulation in normal ventricular muscle. However, the state of NBC in nonischemic hypertrophied hearts is unresolved. In this study, we examined functional and molecular properties of NBC in adult rat ventricular myocytes. The cells were enzymatically isolated from both normal and hypertrophied hearts. Ventricular hypertrophy was induced by pressure overload created by suprarenal abdominal aortic constriction of 50% for 7 wk. pHi was measured in single cells using the fluorescent pH indicator 2′,7′-bis(2-carboxyethyl)5-( 6 )carboxyfluorescein. Real-time PCR analysis was used to quantitatively assess expression of NBC-encoding mRNA, including SLC4A4 (encoding electrogenic NBC, NBCe1) and SLC4A7 (electroneutral NBC, NBCn1). Our results demonstrate that: 1) mRNA levels of both the electrogenic NBCe1 (SLC4A4) and electroneutral NBCn1 (SLC4A7) forms of NBC were increased by aortic constriction, 2) the onset of NBC upregulation occurred within 3 days after constriction, 3) normal and hypertrophied ventricles displayed regional differences in NBC expression, 4) acid extrusion via NBC ( JNBC) was increased significantly in hypertrophied myocytes, 5) although acid extrusion via Na+/H+ exchange was also increased in hypertrophied myocytes, the relative enhancement of JNBC was larger, 6) membrane depolarization markedly increased JNBC in hypertrophied myocytes, and 7) losartan, an ANG II AT1 receptor antagonist, significantly attenuated the upregulation of both NBCs induced by 3 wk of aortic constriction. Enhanced NBC activity during hypertrophic development provides a mechanism for intracellular Na+ overload, which may render the ventricles more vulnerable to Ca2+ overload during ischemia-reperfusion.

Structure and electrical properties of eye muscles in cave- and surface-dwelling crayfishes

1980 ◽  
Vol 84 (1) ◽  
pp. 187-199
Author(s):  
D. Mellon ◽  
G. Lnenicka
Keyword(s):  
Input Resistance ◽  
Selective Advantage ◽  
Oculomotor System ◽  
Membrane Resistance ◽  
Muscle Fibres ◽  
Cross Sectional ◽  
Eye Muscles ◽  
Current Voltage ◽  
The Difference ◽  
Cave Dweller

The morphologies and passive electrical parameters of fibres in two eye muscles of a surface- and a cave-dwelling crayfish were compared. In the cave-dwelling form the muscles contained fewer fibres, of less diameter, and hence had a smaller cross-sectional area. Current-voltage relationships were similar in both species. Input resistance was higher in the cave-dweller, but the difference was not as great as would be expected on the basis of geometry alone. Accordingly, the specific membrane resistance of muscle fibres in the cave-dweller is 50–60% smaller than that in the surface-dweller. This may account partially for the observation that identified excitatory junctional potentials in muscles of cave- and surface dwellers have similar amplitudes. We conclude that a functional oculomotor system is maintained in cave-dwelling crayfish, and that this system confers some positive selective advantage.

Differential contribution of sialic acid to the function of repolarizing K+ currents in ventricular myocytes

2001 ◽  
Vol 281 (2) ◽  
pp. C464-C474 ◽  
Author(s):  
Carmen A. Ufret-Vincenty ◽  
Deborah J. Baro ◽  
L. F. Santana
Keyword(s):  
Sialic Acid ◽  
Voltage Dependence ◽  
Ventricular Myocytes ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Early Afterdepolarizations ◽  
Kv4 Channels ◽  
Ovary Cell Line ◽  
Differential Contribution ◽  
K Currents

We investigated the contribution of sialic acid residues to the K+ currents involved in the repolarization of mouse ventricular myocytes. Ventricular K+ currents had a rapidly inactivating component followed by slowly decaying and sustained components. This current was produced by the summation of three distinct currents: I to, which contributed to the transient component; I ss, which contributed to the sustained component; and I K,slow, which contributed to both components. Incubation of ventricular myocytes with the sialidase neuraminidase reduced the amplitude of I to without altering I K,slow and I ss. We found that the reduction in I to amplitude resulted from a depolarizing shift in the voltage of activation and a reduction in the conductance of I to. Expression of Kv4.3 channels, a major contributor to I to in the ventricle, in a sialylation-deficient Chinese hamster ovary cell line (lec2) mimicked the effects of neuraminidase on the ventricular I to. Furthermore, we showed that sialylated glycolipids have little effect on the voltage dependence of I to. Finally, consistent with its actions on I to, neuraminidase produced an increase in the duration of the action potential of ventricular myocytes and the frequency of early afterdepolarizations. We conclude that sialylation of the proteins forming Kv4 channels is important in determining the voltage dependence and conductance of I to and that incomplete glycosylation of these channels could lead to arrhythmias.

Abstract 3503: Hydrogen Peroxide-Induced Afterdepolarizations are Dependent on Calmodulin Kinase II Signaling

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Lai-Hua Xie ◽  
Fuhua Chen ◽  
James N Weiss
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Ventricular Myocytes ◽  
Heart Association ◽  
Early Afterdepolarizations ◽  
Calmodulin Kinase ◽  
Repolarization Reserve ◽  
Delayed Afterdepolarizations ◽  
Rabbit Ventricular Myocytes ◽  
Camkii Activation

Background: In the heart, hydrogen peroxide (H 2 O 2 ) has been shown to cause early afterdepolarizations (EADs) and triggered activity by impairing Na current (I Na ) inactivation. Since H 2 O 2 has been recently shown to activate Ca 2+ /calmodulin kinase II (CaMKII), and since CaMKII activation has also been reported to impair I Na inactivation and predispose to EADs, we hypothesized that CaMKII activation by H 2 O 2 may be an important factor in the genesis of EADs induced by oxidative stress. Methods and Results: Patch-clamped Fluo-4 AM-loaded rabbit ventricular myocytes were exposed to H 2 O 2 (0.1–1mM), which induced spontaneous EADs after 5–15 min. Both the I Na blocker tetrodoxtin (TTX, 10 μM) and the I Ca,L blocker nifedipine shortened AP duration (APD) and suppressed EADs. H 2 O 2 increased both peak and steady-state I Ca,L under square-pulse voltage clamp, and enhanced I Ca,L to a greater extent during the AP plateau than during the AP upstroke under AP clamp conditions. In addition, by prolonging the AP plateau and increasing Ca influx via maintained I Ca,L , H 2 O 2 -induced EADs frequently caused DADs delayed afterdepolarizations (DADs) due to spontaneous SR Ca release waves after repolarization. KN-93(1 μM), a CaMKII inhibitor, prevented H 2 O 2 -induced EADs (n=4), whereas the inactive analogue KN-92 did not (n=5). Conclusion: These findings indicate that H 2 O 2 -induced EADs depend on both impaired I Na inactivation to reduce repolarization reserve and enhanced I Ca,L to reverse repolarization. Intact CaMKII signaling is necessary for EAD generation in this setting, presumably via its actions on I Na and I Ca,L , although direct redox effects on other ion channels/transporters may also be important. Our observations support a link between increased oxidative stress, CaMKII activation and afterdepolarizations as triggers of lethal ventricular arrhythmias in diseased heart. This research has received full or partial funding support from the American Heart Association, AHA National Center.

Abstract 17019: Two Distinct mechanisms by which Na+/Ca2+ dysregulation contributes to Arrhythmogenic Diastolic Ca2+ Release

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Przemyslaw Radwanski ◽  
Rengasayee Veeraraghavan ◽  
Björn Knollmann ◽  
Sándor Györke
Keyword(s):  
Ventricular Myocytes ◽  
Cellular Level ◽  
Polymorphic Ventricular Tachycardia ◽  
Cell Periphery ◽  
Individualized Approach ◽  
Triggered Arrhythmias ◽  
Local Contribution ◽  
Dependent Protein ◽  
Camkii Activation

Na + and Ca 2+ imbalance is associated with triggered arrhythmias resulting from diastolic Ca 2+ release (DCR) from sarcoplasmic reticulum (SR). Recent evidence suggests Na + channel blockade to be a promising therapy for pathologies, including catecholaminergic polymorphic ventricular tachycardia (CPVT). However, the specific mechanism(s) as to how Na + /Ca 2+ dysregulation contribute to arrhythmias is unknown. Confocal microscopy of ventricular myocytes isolated from CPVT mice lacking the cardiac calsequestrin was used to assess Ca 2+ handling response to isoproterenol (Iso) and various pharmacological interventions, while electrocardiograms were acquired during catecholamine challenge to assess the roles of various pools of Na + channels in CPVT. We identify two pools of Na + channels: one composed of cardiac-type Na + channels localized to cell periphery, and a ‘ local pool ’ comprised of neuronal Na + channels colocalizing with RyR2 in the T-tubules. Augmenting function of both Na + channel pools with ATX-II in the presence Iso resulted in SR Ca 2+ overload and activation of Ca 2+ /calmodulin-dependent protein kinase II (CaMKII), which precipitated DCR. These, in turn, translated into frequent arrhythmias in CPVT mice. Selectively augmenting function of ‘local pool’ neuronal Na + channels with β-Pompilidotoxin (β-PMTX) precipitated DCR on the cellular level causing frequent arrhythmias during catecholamine challenge in vivo . However, increasing local Na + fluxes reduced SR Ca 2+ load suggesting that local elevation in cytosolic Ca 2+ rather than global SR Ca 2+ overload underlies DCR and arrhythmias under such conditions. These data suggest two distinct mechanisms for Na + /Ca 2+ dysregulation-mediated arrhythmias. The first relies on SR Ca 2+ overload and CaMKII activation and the other on local contribution of Na + -Ca 2+ exchange to DCR. Consideration of these divergent mechanisms may enhance individualized approach to arrhythmia management.

Effects of ouabain on intracellular ion activities of sensory neurons of the leech central nervous system

1991 ◽  
Vol 65 (3) ◽  
pp. 736-746 ◽  
Author(s):  
W. R. Schlue
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Input Resistance ◽  
Membrane Resistance ◽  
Membrane Potential Change ◽  
Intracellular Na Activity ◽  
Order Of Magnitude ◽  
Intracellular Ion Activities ◽  
Ion Activities ◽  
K Concentration

1. The intracellular K+, Na+, and Ca2+ of mechanosensory neurons in the central nervous system of the leech Hirudo medicinalis was measured using double-barreled ion-sensitive microelectrodes. 2. After inhibition of the Na(+)-K+ pump with 5 x 10(-4) M ouabain, the intracellular K+ activity (aKi) decreased, while the intracellular Na+ activity (aNai) increased. The input resistance decreased in the presence of ouabain. The intracellular Ca2+ increased more than one order of magnitude after ouabain addition. All changes in intracellular ion activities and membrane resistance were fully reversible. 3. When extracellular Na+ concentration ([Na+]o) was removed [replaced by tris(hydroxymethyl)aminomethane (Tris)], aNai decreased. In the absence of [Na+]o, aKi and aNai remained unchanged after inhibition of the Na(+)-K+ pump by reducing the extracellular K+ concentration ([K+]o) to 0.2 mM. The membrane resistance increased under these conditions. 4. The intracellular Ca2+ decreased or remained constant after removal of [Na+]o. Addition of ouabain in the absence of [Na+]o did not change intracellular Ca2+, which only increased after readdition of [Na+]o. 5. The relative K+ permeability (PK) measured as membrane potential change during a brief increase of the [K+]o from 4 to 10 mM, increased manyfold after addition of ouabain but only little if [Na+]o had been removed before adding ouabain. 6. The results suggest that the intracellular Na+ increase after inhibition of the Na(+)-K+ pump affects the intracellular Ca2+ level by stimulating a Nai(+)-Ca2+ exchange mechanism. The subsequent intracellular Ca2+ activity (aCai) rise may result in an increase of the membrane permeability to K+ ions.

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