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S L Lipsius

Publications and source records attributed to S L Lipsius.

At least 37 records · Page 2Linked to original sources
PubMed1995

Acetylcholine elicits a rebound stimulation of Ca2+ current mediated by pertussis toxin-sensitive G protein and cAMP-dependent protein kinase A in atrial myocytes.

Cholinergic inhibition of atrial contraction is typically followed by a rebound positive inotropic response. In the present study, we used a nystatin-perforated patch whole-cell recording method to determine whether acetylcholine (ACh) elicits a rebound stimulation of L-type Ca2+ current (ICa,L) in cat atrial myocytes. ACh (1 mumol/L) decreased basal ICa,L (-19 +/- 2%). Within approximately 30 s of returning to ACh-free solution, basal ICa,L exhibited a rebound increase above the control level (+61 +/- 7%) that returned to the control level within 4 to 5 minutes. ACh elicited concomitant changes in cell shortening, ie, a decrease followed by a rebound increase. The EC50 and maximal response of ACh-induced inhibition and rebound stimulation of ICa,L were 1.9 x 10(-9) mol/L and -30%, respectively, and 2.9 x 10(-8) mol/L and +64%, respectively. All effects of ACh on ICa,L were blocked by prior exposure to 1 mumol/L atropine or 100 mumol/L AFDX116 and unaffected by 0.2 mumol/L pirenzepine or 1 mumol/L propranolol. In the presence of ACh, exposure to atropine elicited stimulation of ICa,L.ACh-induced inhibition and rebound stimulation of current were independent of external Ca2+. Rebound stimulation of ICa,L was associated with a negative shift in the voltage dependence of ICa,L activation. Inhibition of protein kinase A by 50 mumol/L Rp-cAMPs decreased basal ICa,L by 36 +/- 1% and abolished the rebound stimulation of ICa,L. Forskolin (0.01 mumol/L) or isoproterenol (0.01 mumol/L) had no effect on basal ICa,L, but each accentuated the rebound increase in ICa,L. When adenylate cyclase was maximally stimulated with 1 mumol/L isoproterenol plus 2 mumol/L forskolin, ACh decreased ICa,L but failed to elicit rebound stimulation of ICa,L. Milrinone (10 mumol/L) increased basal ICa,L by 70 +/- 7% and significantly attenuated the rebound stimulation of ICa,L. Exposure to 1 mmol/L 8-bromo-cGMP elicited a small decrease in basal ICa,L, attenuated ACh-induced inhibition, and enhanced the rebound stimulation of ICa,L. Incubation in pertussis toxin prevented all ACh-induced changes in ICa,L. Inhibition of nitric oxide synthase by 100 mumol/L NG-monomethyl-L-arginine (L-NMMA) decreased basal ICa,L by -20 +/- 5%, prevented ACh-induced inhibition, and markedly attenuated the rebound stimulation of ICa,L. We conclude that in cat atrial myocytes ACh acts via M2 muscarinic receptors and pertussis toxin-sensitive G protein to inhibit basal ICa,L and that on withdrawal ACh elicits a rebound stimulation of ICa,L. Rebound stimulation of ICa,L is mediated via cAMP-dependent protein kinase A enhanced by ACh-induced inhibition of phosphodiesterase.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine↗
PubMed1995

beta-Adrenergic stimulation induces acetylcholine to activate ATP-sensitive K+ current in cat atrial myocytes.

Our previous work on atrial myocytes suggested that the effect of acetylcholine (ACh) to increase K+ conductance can be potentiated by prior loading of the sarcoplasmic reticulum (SR) with Ca2+. The present study, therefore, sought to determine whether prior exposure to isoproterenol (ISO) potentiates ACh-induced increases in K+ conductance and the underlying mechanisms. A nystatin-perforated patch whole-cell configuration was used to record from cat atrial myocytes. Voltage-clamp ramps (40 mV/s) were used to assess total membrane conductance. The experimental protocol consisted of two consecutive 30-second ACh exposures (ACh1 and ACh2) separated by a 6-minute recovery period in ACh-free solution. In general, experimental interventions, such as exposure to ISO, were imposed during the period between ACh1 and ACh2 to determine their effects on the response to ACh2 in relation to ACh1. Under control conditions, K+ conductances induced by ACh1 and ACh2 were not different from one another with or without activation of L-type Ca2+ current (ICa,L) during the recovery period. When 1 mumol/L ISO plus ICa,L activation was imposed during the recovery period, ACh2 induced a significantly larger increase in K+ conductance than ACh1. The ACh2-induced K+ current potentiated by ISO was time independent and selectively blocked by 10 mumol/L glibenclamide and therefore identified as ATP-sensitive K+ current (IK,ATP). The effect of ISO to induce ACh2-activated IK,ATP was mimicked by 1 mumol/L forskolin or 200 mumol/L 8-(4-chlorophenylthio)-cAMP, but not by 0.5 mumol/L BAY K 8644, and was selectively abolished by (1) 5 mumol/L thapsigargin or 1 mumol/L ryanodine, agents that prevent accumulation of SR Ca2+, (2) inhibition of L-type Ca2+ current (ICa,L) by 1 mumol/L nisoldipine or zero external Ca2+, (3) 50 mumol/L Rp-cAMPs, an inhibitor of cAMP-dependent protein kinase A, or (4) 2 mumol/L propranolol. Atropine (1 mumol/L) abolished all ACh-induced currents. Moreover, ACh2-activated IK,ATP was selectively blocked by 0.2 mumol/L pirenzepine, an M1 muscarinic receptor antagonist, or 0.1 mumol/L calphostin C, a selective inhibitor of protein kinase C. AFDX116 (100 mumol/L), an M2 muscarinic receptor antagonist, blocked the conventional ACh-activated K+ current and revealed ACh2-activated IK,ATP. These results indicate that prior exposure to ISO potentiates ACh-induced increases in K+ current via ACh-activated IK,ATP.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine↗
PubMed1994

T-type calcium current in latent pacemaker cells isolated from cat right atrium.

Whole-cell voltage clamp techniques were used to study Ca2+ currents in latent pacemaker cells isolated from cat right atrium. T-type (ICa,T) and L-type (ICa.L) Ca2+ currents were distinguished by their voltage dependence of activation, and sensitivity to channel blocking agents. In 2.7 mM [Ca]o, ICa,T activation exhibited a voltage threshold of about -50mV and maximum amplitude at -10mV, whereas ICa,L threshold was about -30mV and maximum amplitude was at +10mV. The half-maximal activation voltages of ICa,T was -31.4 +/- 0.2mV and ICa,L was -6.2 +/- 2.0mV. Overlap of the steady-state activation-inactivation curves for ICa,T showed a "window" current at voltages compatible with the late phase of diastolic depolarization. Maximum ICa,T and ICa,L current densities were 3.3 +/- 0.4pA/pF and 12.5 +/- 1.3pA/pF, respectively. ICa,T current density in working atrial muscle cells was 0.73 +/- 0.31pA/pF. Both ICa,L and ICa,T were blocked by 2mM cobalt. ICa,L but not ICa,T was blocked by 1 microM nifedipine. Nickel (Ni2+; 40 microM) inhibited ICa,T primarily at more negative voltages. In addition, Ni2+ decreased the late phase of diastolic depolarization and significantly increased pacemaker cycle length. These results indicate that latent atrial pacemaker cells exhibit ICa,T that is significantly larger in amplitude than in working atrial muscle cells. ICa,T may contribute current during the late phase of diastolic depolarization. Because latent pacemakers exhibit a more negative maximum diastolic potential, ICa,T may contribute more to latent than to primary pacemaker activity.

Animals↗
PubMed1994

Delayed rectifier potassium current (IK) in latent atrial pacemaker cells isolated from cat right atrium.

Whole-cell recording techniques were used to study the delayed rectifier K+ current (IK) in latent pacemaker cells isolated from cat right atrium. From a holding potential of -40 mV, depolarizing clamp steps elicited L-type Ca2+ current followed by an increasing outward current (IK). The time course of tail current amplitudes paralleled that of the time-dependent activation of outward current. Activation of IK exhibited a sigmoidal time course that was best fit by a power function where the activation variable was raised to the second power. The voltage-dependence of IK activation exhibited a sigmoidal relationship between -40 and +30 mV. The half-maximal activation voltage and slope factor were -21.9 +/- 1.3 and 13.8 +/- 0.9 mV respectively (n = 6). The fully activated I/V relationship of IK was linear between -100 and -30 mV and inwardly rectified at more positive voltages. Following IK activation, hyperpolarizations more negative than about -50 mV elicited tail currents that consisted of both IK deactivation and I(f) activation. A subtraction protocol was used to isolate IK tail currents. In 5.4 mM extracellular [K+], IK tail currents exhibited a reversal potential of -78.2 +/- 0.3 mV (n = 6). The reversal potential of IK was linearly related to log extracellular [K] and the slope was 51.5 mV per ten-fold change in extracellular [K]. At -70 mV, IK tail currents decayed as a single exponential function with a time constant of 159 +/- 16 ms (n = 6). These results indicate that latent atrial pacemakers exhibit IK activated by depolarization.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1993

Effect of isoprenaline on I(f) current in latent pacemaker cells isolated from cat right atrium: ruptured vs. perforated patch whole-cell recording methods.

Whole-cell recording techniques were used to study the time-dependent inward current activated on hyperpolarization (I(f)) and its response to isoprenaline in latent atrial pacemaker cells isolated from cat right atrium. To determine whether the response to isoprenaline depended on the type of recording method, we analysed I(f) using either a standard ruptured-patch, or a nystatin-perforated-patch, whole-cell recording method. All cells beat rhythmically at 35 degrees C and exhibited normal pacemaker action potentials and I(f) current, regardless of the recording method. With the ruptured-patch method, pacemaker action potentials ceased activity within a few minutes and I(f) amplitude decreased "ran down" to 74% of control within 10 min of rupturing the patch. Isoprenaline (1 microM) elicited variable changes in I(f) amplitude among different latent pacemaker cells resulting in no net change in mean current amplitude (n = 6). In addition, isoprenaline failed to change the voltage dependence of the I(f) activation curve. On the other hand, using a nystatin-perforated-patch method, pacemaker action potentials and I(f) exhibited no significant changes over the same 10 min period. Under these conditions, isoprenaline consistently increased I(f) in all cells studied (+90%) at -80 mV; n = 8), and increased the spontaneous rate of pacemaker action potentials by 58 +/- 7% (n = 5). Moreover, isoprenaline elicited a significant positive shift (+11 mV) in the half-maximal activation voltage of the I(f) activation curve (n = 3). We conclude that latent atrial pacemakers consistently exhibit I(f) current, and that isoprenaline consistently elicits an increase in I(f) amplitude.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1993

Phase-dependent properties of the cardiac sarcoplasmic reticulum oscillator in cat right atrium: a mechanism contributing to dysrhythmias induced by Ca2+ overload.

These experiments analyse the phase-dependent properties of spontaneous oscillations of the sarcoplasmic reticulum (SR) induced by Ca2+ overload. Right atrial tissue was loaded with intracellular Ca2+ by exposure to a modified Tyrode solution containing 50% of normal Na+ and 0.5 mM K+. Verapamil (2 microM) was added to block regenerative activity. Intracellular Ca2+ overload elicited spontaneous, rhythmic voltage and tension oscillations that were phase locked 1:1. Voltage and tension oscillations were abolished by exposure to low (0.9 mM) external Ca2+, 1 microM ryanodine, or 10 mM caffeine, indicating that both voltage and tension oscillations resulted from spontaneous oscillations in SR Ca2+ release. Single pulses of nerve-stimulated ACh release elicited phase shifts in both voltage and tension oscillations. Sinusoidal current was used as a periodic stimulus to drive membrane voltage and elicit periodic voltage oscillations. Stimulated voltage oscillations entrained spontaneous tension oscillations 1:1 in a range of frequencies close to the basic spontaneous SR oscillatory cycle length, or 2:1 at frequencies close to one-half the spontaneous SR oscillatory cycle length. Stimulation frequencies between these two regions entrained tension oscillations in predictable fixed coupled ratios (4:3, 3:2) and resulted in Wenckeback-like voltage patterns. Stimulation frequencies between phase-locked regions resulted in complex coupling relationships and irregular voltage patterns. Exposure to 1 microM ryanodine, 0.9 mM external Ca2+, or 10 mM caffeine abolished irregular voltage patterns and tension. We conclude that the SR oscillator exhibits phase-dependent sensitivity to perturbations at the surface membrane. As a result, external perturbations can elicit phase differences between spontaneous SR oscillations and membrane voltage that cause either phase-locked or irregular voltage patterns. These findings identify an intracellular mechanism that may contribute to the development of cardiac dysrhythmias resulting from intracellular Ca2+ overload.

Acetylcholine↗
PubMed1993

Na(+)-Ca2+ exchange current in latent pacemaker cells isolated from cat right atrium.

1. Single latent pacemaker cells were isolated from cat right atrium, and studied in a whole-cell configuration using a nystatin-perforated patch recording method. The nystatin method avoids alterations in intracellular Ca2+, cellular constituents and run-down of ionic currents. 2. Depolarizing voltage clamp pulses from -40 mV elicited L-type Ca2+ current (ICa) that exhibited an initial rapid phase of inactivation followed by a secondary slower inward current component that decayed over about 100 ms. The secondary inward component appeared as a slowly decaying inward tail current following short (10-40 ms) depolarizing clamp steps. 3. Slowly decaying inward currents were abolished by internally dialysing pacemaker cells with 2 mM EGTA using a ruptured patch recording method. Inward tail currents were also abolished by exposure to 1 microM ryanodine and significantly decreased by replacing 85% of external Na+ with lithium, without effect on peak ICa. These findings identify a Na(+)-Ca2+ exchange current (INa-Ca) that is mediated by sarcoplasmic reticulum (SR) Ca2+ release. 4. Properties of INa-Ca and ICa differed significantly: (i) ICa exhibited a bell-shaped voltage dependence that peaked at 0 mV and decreased at more positive voltages. INa-Ca was maximal at -10 mV and remained relatively constant at more positive voltages; (ii) a paired pulse protocol showed that the time course of INa-Ca recovery (5 s) was significantly longer than that of ICa (2 s); (iii) cadmium (50 microM) induced an inhibition of ICa that did not correlate in time with changes in INa-Ca. 5. The duration of depolarizing steps between 10 and 120 ms had no effect on the time course of INa-Ca tail currents. 6. Isoprenaline > or = 5 x 10(-8) M significantly increased peak ICa amplitude, peak INa-Ca amplitude, accelerated INa-Ca rate of decay and decreased the absolute time of INa-Ca decay. 7. Free-running pacemaker action potentials were clamped during diastole at either -40 or -70 mV (maximum diastolic potential) for variable periods of time. At times between 0.2 and 1 s, INa-Ca exhibited a voltage-dependent increase in amplitude over time, i.e. INa-Ca recovered more rapidly from -70 mV than from -40 mV. At times > 2 s, INa-Ca exhibited a voltage-dependent decline in amplitude over time, i.e. from -40 mV INa-Ca decreased by 10% of maximum whereas from -70 mV INa-Ca decreased by 60% of maximum.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1992

Properties of the pacemaker current (If) in latent pacemaker cells isolated from cat right atrium.

1. Single latent pacemaker cells were isolated from the Eustachian ridge of cat right atrium using Langendorff perfusion and enzyme dispersion techniques. Whole-cell patch-clamp techniques were used to study the hyperpolarization-activated inward current (I(f)). 2. All cells studied beat rhythmically. Pacemaker activity was recorded in the voltage range -68 +/- 1 to -54 +/- 2 mV and its cycle length was 901 +/- 67 ms (72 +/- 5 beats min-1) at 34-36 degrees C. Cells were elongated with tapered ends, and appeared bent or crinkled without obvious striations. Mean cell diameter and length were 7.4 +/- 0.5 microns and 93.1 +/- 5.9 microns, respectively (n = 15). Input resistance and total membrane capacitance were 2.2 +/- 0.2 G omega and 27.8 +/- 3.1 pF, respectively. 3. Hyperpolarizing clamp steps more negative than -50 mV elicited a time-dependent increasing inward current that was maximally activated at -120 mV. Activation of I(f) was well within the pacemaker voltage range. Half-maximal activation voltage and slope factor were calculated, using a Boltzmann function, to be -80.5 mV and 8.4, respectively. 4. The fully activated current-voltage (I-V) relationship was approximately linear at voltages more negative than -30 mV and showed outward rectification at more positive voltages. The reversal potential of I(f) was -26 mV and the fully activated conductance was 1.75 +/- 0.14 nS (n = 21). Caesium (2 mM) blocked I(f) at voltages more negative than the reversal potential. Reducing extracellular Na+ or K+ shifted the reversal potential more negative, and increasing extracellular K+ exerted the opposite effect. Reducing extracellular Na+ decreased I(f) amplitude and the slope of the fully activated I-V relationship, and elevated extracellular K+ increased I(f) amplitude and the slope of the fully activated I-V relationship. 5. Some pacemaker cells exhibited a short delay in the onset of I(f) activation whereas other pacemaker cells exhibited little, if any, delay in activation. I(f) currents exhibiting no delay in activation were best fitted by a single exponential function with a mean time constant of 3.20 +/- 1.03 s at -70 mV (n = 4). 6. A nystatin-permeabilized patch recording method was used to record spontaneous pacemaker action potentials and I(f) from the same pacemaker cell. Caesium (2 mM) inhibited I(f) by more than 90% (at -70 mV), and decreased the slope of diastolic depolarization, resulting in a 48 +/- 5% decrease in spontaneous rate.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1991

Ionic currents activated during hyperpolarization of single right atrial myocytes from cat heart.

Whole-cell recording techniques were used on single right atrial myocytes to study the ionic currents that may be responsible for the diverse diastolic voltage characteristics of atrial tissue. Ionic currents were activated by hyperpolarizing voltage pulses negative to -30 mV. In general, four different types of cells were identified based primarily on the ionic currents elicited during hyperpolarization. The first cell type exhibited an inward current that decayed with time at more negative voltages, reversed near the potassium equilibrium potential, inwardly rectified at more positive voltages, increased in elevated extracellular potassium, and was blocked by 3 mM barium or 10 mM cesium. This current was identified as the potassium current iK1. A second cell type exhibited a time-dependent inward current that increased at more negative voltages, had an activation range between -50 and -110 mV, had a reversal potential of -26 mV, and was blocked by 3 mM cesium. This current was identified as an if current. A third cell type exhibited an inward current that initially decayed and then became more inward with time. Barium (3 mM) abolished the initial inward current and revealed a time-dependent increasing inward current that was blocked by 3 mM cesium. This current was composed of both the iK1 and if currents. A fourth cell type exhibited only small time-independent leak currents in response to hyperpolarization. These results indicate that individual cells within the right atrium are electrophysiologically heterogeneous with respect to the types of ionic channels present in their sarcolemmal membranes. This specialization in ionic currents partially explains the diverse diastolic voltage characteristics and functional properties of atrial tissue.

Action Potentials↗
PubMed1990

Effects of extracellular Mg2+ on T- and L-type Ca2+ currents in single atrial myocytes.

Whole cell voltage-clamp techniques were used to study the effects of extracellular Mg2+ on T- and L-type Ca2+ currents recorded from single atrial myocytes from cat heart. T and L currents were distinguished primarily by their voltage dependence. With 5.4 mM Ca2+ as charge carrier, maximal T- and L-current densities were 1.0 +/- 0.06 and 9.7 +/- 0.4 pA/pF, respectively. Nickel (Ni2+, 50 microM) inhibited maximal T current (-65.6 +/- 5.9%) more than L current (-15.7 +/- 2.4%), and 10 microM cadmium (Cd2+) inhibited L current (-65.5 +/- 5.9%) without significant effect on T current (-8.7 +/- 8.1%). Mg2+ elicited a dose-dependent inhibition of both T and L currents. Mg2+ less than 8.4 mM inhibited T current more than L current. At Mg2+ greater than or equal to 8.4 mM, T-current inhibition reach a plateau at approximately 52%, whereas L current was further inhibited (-65%) at 16.8 mM Mg2+. Mg2+ elicited a dose-dependent positive shift in half-maximal voltages of activation and inactivation for both T and L currents. Mg2(+)-induced inhibition of both T and L currents was greater in lower (2.7 mM) external Ca2+. Finally, 4.2 mM Mg2+ and 50 microM Ni2+ elicited a similar decrease in the late diastolic slope of subsidiary pacemaker action potentials, whereas 10 microM Cd2+ markedly inhibited action potential amplitude.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1989

Mechanisms of automaticity in subsidiary pacemakers from cat right atrium.

Intracellular recordings were made from eustachian ridge of cat right atrium to determine mechanisms responsible for subsidiary pacemaker automaticity. Pacemaker action potentials exhibited two phases of diastolic depolarization: an initial steeper slope (D1) followed by a more gradual slope (D2). Cesium (1 mM) decreased D1 (-45.6%) to a significantly greater extent than D2 (-33.6%) and increased spontaneous cycle length (SCL) (+37.7%). Tetrodotoxin (10(-6) M) had no effect on maximum rate of rise of upstroke, although it increased SLC (+23.9%). Verapamil (0.4-1.0 microM) progressively increased SCL by decreasing late diastolic slope, resulting in oscillatory potentials and eventual quiescence. Both norepinephrine (2 x 10(-9) M) and Bay K 8644 (10(-7) M) elicited a significantly greater increase in D2 than in D1, resulting in a decrease in SCL. Ryanodine (10(-6) M) caused a small but significant initial decrease (-3.7%) followed by a progressive increase in SCL (+172%). Ryanodine decreased D2 without changing D1, increased maximum rate of rise and overshoot potential, and abolished tension. In the presence of ryanodine, Bay K 8644 progressively increased D1 amplitude, resulting in a cyclic pattern of dysrhythmic activity. In the presence of ryanodine, cesium significantly decreased D1 (-39.3%), shifted the late diastolic potential more negative, and increased SCL (+25.7%). These results indicated that multiple mechanisms participate in subsidiary pacemaker automaticity. They include 1) a cesium-sensitive component that contributes to a greater extent during the initial phase of diastolic depolarization, 2) a component mediated via calcium released from the sarcoplasmic reticulum that contributes primarily during the latter half of diastolic depolarization, and 3) possibly a direct contribution by the slow inward calcium current.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1987

Triggered rhythms in atrial muscle.

Intracellular and extracellular recording techniques were used to study triggered activity in tissues isolated from the inferior right atrium of the dog. In the presence of norepinephrine (greater than 10(-7) M) single stimuli or short rapid pacing elicited action potentials with delayed after-depolarizations, leading to single or repetitive non-driven beats. Most commonly, during sustained triggered activity action potential and electrogram configurations remained constant. In addition, cycle length changed gradually, initially decreasing and then increasing before activity stopped. However in some preparations, once sustained triggered activity was initiated there were abrupt spontaneous changes in action potential and electrogram configurations that were coincident with abrupt spontaneous changes in cycle length. When quiescent, short rapid pacing initiated triggered activity with a relatively short cycle length that abruptly shifted to a longer cycle length before stopping. During sustained triggered activity, short rapid pacing caused an abrupt decrease in cycle length that was coincident with changes in action potential and electrogram configurations. The present results indicate that abrupt changes in cycle length can occur during bouts of triggered activity resulting in unique patterns of arrhythmias. These rhythms may be due to interactions and shifts between multiple sites of triggered pacemaker activity.

Action Potentials↗
PubMed1987

Electrophysiology and ultrastructure of eustachian ridge from cat right atrium: a comparison with SA node.

Intracellular microelectrode and electron microscopic techniques were used to investigate and correlate the electrophysiology of subsidiary pacemaker activity with the presence of cells having ultrastructural characteristics of pacemaker cells i.e. P cells, in Eustachian ridge tissue isolated from cat right atrium. In addition, the electrophysiological characteristics of subsidiary pacemaker activity and the ultrastructural characteristics of P cells in Eustachian ridge were compared to those of SA node obtained from the same hearts. Action potential recordings and morphological analysis were restricted to the endocardial site of earliest activation. Electrophysiological recordings revealed that spontaneously active Eustachian ridge tissues generate slow response action potentials with pacemaker characteristics similar, although not identical, to those of SA node. These included a relatively steep diastolic slope, low maximum diastolic potential (-70 mV), rate of rise (5.5 V/s), take-off potential (-52.5 mV), a relatively large overshoot potential (+7.7 mV) and a spontaneous cycle length (948 ms) about twice as long as SA node (434 ms). Morphological analysis revealed cells with ultrastructural characteristics of P cells, that were restricted to the endocardial site of earliest pacemaker activation. Morphological measurements indicate that Eustachian ridge P cells are not significantly different from P cells in SA node of the same hearts. However, Eustachian ridge P cells exhibit a unique apposition of subsarcolemmal cisternae between cells not seen in SA node. We conclude that pacemaker cells within the Eustachian ridge generate stable, spontaneous activity via slow response pacemaker action potentials. Cells responsible for this subsidiary pacemaker activity are most likely P cell types that are similar, although not identical, to P cells in SA node.

Action Potentials↗
PubMed1985

Electrophysiology of functional subsidiary pacemakers in canine right atrium.

Glass microelectrodes were used to study the electrical activity of the subsidiary atrial pacemaker (SAP) cells that maintain atrial excitation after suppression of the sinoatrial node. Tissues with documented SAP activity were isolated from the canine inferior right atrium and superfused in vitro with Tyrode solution containing norepinephrine (NE, 10(-8)-10(-7) M). SAP action potentials exhibited prominent diastolic depolarization and a significantly lower maximum diastolic potential, take-off potential, overshoot, rate of rise, and amplitude than typical atrial muscle. Withdrawal of NE completely blocked SAP propagation, although SAP automaticity continued at a slower rate. Acetylcholine (ACh, 5 X 10(-8) M) usually produced complete exit block and decreased spontaneous rate. Higher concentrations of ACh (10(-6) M) elicited a prominent hyperpolarization (19.2 +/- 6.6 mV), completely suppressing SAP automaticity. In quiescent preparations exposed to NE greater than or equal to 10(-7) M, external stimuli at short cycle lengths (less than 1,000 ms) elicited action potentials with delayed afterdepolarizations, which frequently caused nondriven repetitive activity. This triggered activity was inhibited by verapamil or withdrawal of NE. These studies identify and characterize the electrical activity of functional subsidiary pacemakers located in a specific region of the inferior right atrium. In addition, fibers within this region display triggered activity. Spontaneous activity generated by fibers within the SAP region may cause atrial dysrhythmias.

Acetylcholine↗
PubMed1984

Alterations in subsidiary pacemaker function after prolonged subsidiary pacemaker dominance in the canine right atrium.

In vivo and in vitro techniques were used to study the functional characteristics of subsidiary atrial pacemaker activity 4 to 11 months after surgical excision of the sinoatrial node region of the dog heart. Characteristics of this long-term subsidiary atrial pacemaker activity were compared with those of sinoatrial node and short-term subsidiary atrial pacemaker activities. Extracellular bipolar electrodes were used to estimate the site of earliest activation and monitor spontaneous rate. Under both in vivo and in vitro conditions, long-term pacemaker activity was located in the same region of the inferior right atrium as was short-term pacemaker activity. Under in vitro conditions, long-term activity was characterized by a reduced sensitivity to acetylcholine and overdrive pacing and by a lack of dependence on beta-adrenergic stimulation compared with short-term activity. Furthermore, long-term pacemaker activity was more sensitive to acetylcholine and less sensitive to norepinephrine (greater than 10(-7) M) compared with sinoatrial node activity. It is concluded that the subsidiary atrial pacemakers that emerge soon after removal of the sinoatrial node are the same as those pacemakers that ultimately assume long-term control of the heart. In addition, after subsidiary atrial pacemakers assume dominant pacemaker function, their dependence on norepinephrine and their response to acetylcholine and overdrive pacing are reduced. These changes facilitate a more stable regulation of atrial pacemaker rhythm and, thereby, make subsidiary atrial pacemaker function more like that of the sinoatrial node.

Acetylcholine↗
PubMed1983

Electrotonic interactions in delayed propagation and block within the guinea pig SA node.

The influence of electrotonic interactions on propagation within the SA node was studied by recording transmembrane potentials simultaneously from two neighboring (less than 1 mm apart) subsidiary pacemaker cells within the sinoatrial (SA) node of the guinea pig. As single premature stimuli were delivered progressively earlier in diastole, retrograde propagation between cells was delayed progressively. Cells activated earlier displayed secondary depolarizations that were coincident with the depolarization of neighboring cells activated later. The secondary depolarizations increased action potential duration markedly. Rapid pacing elicited secondary depolarizations that resulted in a progressive increase in action potential duration and decrease in upstroke amplitude. These changes were associated with a progressive delay in retrograde propagation that led to intermittent block with Wenckebach periodicity. Exposure to tetrodotoxin (10(-5) g/ml) delayed antegrade propagation, resulting in electrotonically mediated secondary depolarizations and exit block with Wenckebach periodicity. It is concluded that delayed activation and electrotonically mediated interactions between cells can increase action potential duration and refractoriness. These changes contribute to progressive delays in propagation that may result in intermittent block with Wenckebach periodicity within the SA node.

Action Potentials↗
PubMed1983

Functional characteristics of sinoatrial and subsidiary pacemaker activity in the canine right atrium.

A canine in vitro right atrial preparation was developed to study the functional characteristics of subsidiary atrial pacemaker (SAP) activity and to compare them with those of sinoatrial node (SAN) activity. Extracellular bipolar electrodes were used to estimate the site of earliest activation and monitor spontaneous rate. Ligation of the SAN artery at the midportion of the sulcus terminalis suppressed SAN activity and usually (73.5%) elicited SAP activity in a well-defined region of the inferior atrium. SAP activity in this region required a "threshold" concentration of norepinephrine (10(-8) M) in the Tyrode's perfusate. In response to all concentrations of norepinephrine tested, SAN activity attained a greater maximum spontaneous rate than SAP activity. Cholinergic stimulation with acetylcholine or eserine elicited a greater negative chronotropic response from SAP than SAN activity. Overdrive pacing suppressed SAP activity to a significantly greater extent than SAN activity. We conclude that this in vitro preparation can be useful for studying the pharmacology and electrophysiology of subsidiary atrial pacemakers that emerge after suppression of SAN activity. In contrast to SAN activity, SAP activity requires norepinephrine and is more sensitive than SAN activity to acetylcholine and overdrive pacing. Consequently, after loss of SAN function, autonomic modulation of SAP activity may result in atrial dysrhythmias and prolonged periods of overdrive suppression.

Acetylcholine↗