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S S Sheu

Publications and source records attributed to S S Sheu.

At least 55 records · Page 3Linked to original sources
PubMed1990

Modulation of ventricular action potential by alpha 1-adrenoceptors and protein kinase C.

The effects of the alpha 1-adrenoceptor agonist methoxamine (MTX) and the direct protein kinase C (PKC) activator phorbol 12,13-dibutyrate (PDBU) on action potentials from guinea pig papillary muscles were studied. Measured with conventional microelectrodes, MTX (1 x 10(-7) to 3 x 10(-4) M) and PDBU (1 x 10(-9) to 1 x 10(-6) M) both caused a dose-dependent and reversible decrease in the action potential duration measured at 90% repolarization (APD90) at 36.5 degrees C. The MTX-mediated response was blocked by both prazosin (3 x 10(-6) M) and phentolamine (1 x 10(-6) M) and mimicked by phenylephrine. Maximal concentrations of the two agents together resulted in only a partial (50%) additive decrease in the APD90. At a lower temperature (27.5 degrees C), PDBU no longer produced a shortening in the APD90 and MTX produced a prolongation in the APD90. These results demonstrate that although alpha 1-adrenoceptor stimulation and PKC activation in guinea pig papillary muscle both lead to a decrease in the APD90, the differences in their effects as related to the magnitude, partial additivity, and temperature dependence suggest that the mechanism of action are not identical. These subtle differences may help to delineate the exact physiological implications of alpha 1-adrenoceptors in cardiac excitation and contraction.

Action Potentials↗
PubMed1990

Spatial heterogeneity of intracellular Ca2+ concentration in nonbeating guinea pig ventricular myocytes.

The spatial distribution of intracellular Ca2+ concentration was determined by fluorescent digital imaging microscopy in fura-2-loaded quiescent cardiac myocytes isolated from guinea pig ventricle. Fluorescent ratio images revealed discrete as well as clustered bright fluorescent spots ("hot spots"), which occupied approximately 20-50% of an individual cell's area. The fluorescent intensity and the area of the hot spots were increased by agents that deplete Ca2+ in the sarcoplasmic reticulum, namely, ryanodine (20-40 nM) and caffeine (5-15 mM). However, when cells were exposed to agents that deplete mitochondrial Ca2+, such as the protonophore, carbonyl cyanide m-chlorophenyl-hydrazone (CCCP, 100-300 nM), or the inhibitor of electron transport, antimycin A (4-40 nM), the fluorescent intensity and the area of the hot spots were reduced. These results indicate that the spatial distribution of intracellular Ca2+ concentration in the ventricular myocytes of guinea pig is quite heterogeneous. The ability of CCCP and antimycin A, but not of caffeine and ryanodine, to reduce the fluorescent intensity in the hot spots implies that Ca2+ compartmentation in the mitochondria is largely responsible for the intracellular Ca2+ heterogeneity seen in the present study.

Animals↗
PubMed1990

Antipsychotic pimozide is a potent Ca2+ channel blocker in heart.

The diphenylbutylpiperidine (DPBP) antipsychotic pimozide was identified as a potent new Ca2+ channel antagonist in heart. In whole-cell patch-clamp experiments, pimozide blocked Ca2+ current through L type channels of rat ventricular myocytes, in a voltage-dependent manner. At holding potentials positive to -40 mV, approximately 90% of current was blocked by 200 nM pimozide. Nearly half of this block, 48 +/- 5% (mean +/- SE, n = 5), was relieved by 5-min hyperpolarization to -100 mV. In quin2-loaded myocytes, pimozide blocked 50 mM KCl-induced increases in intracellular Ca2+ concentration [( Ca2+]i) half maximally at a concentration of 75 +/- 15 nM (n = 5). Two other DPBPs, penfluridol and fluspirilene, also blocked the KCl-induced response at similar concentrations. The contractile force of cardiac tissue was also depressed by pimozide. One micromolar pimozide reduced twitch tension in rat papillary muscles by an average of 36 +/- 8% (n = 3). These results demonstrate that the DPBPs comprise a potent new class of Ca2+ antagonists in heart, which will be useful in studying cardiac Ca2+ channels. They also suggest that these agents may have therapeutic value outside the field of psychiatry.

Aminoquinolines↗
PubMed1990

Perturbations of intracellular calcium distribution in kidney cells by nephrotoxic haloalkenyl cysteine S-conjugates.

The Ca2(+)-sensitive dye fura-2 was used to investigate the disturbances in intracellular Ca2+ concentration [( Ca2+]i) and distribution induced by the nephrotoxic cysteine S-conjugate S-(1,2-dichlorovinyl)-L-cysteine (DCVC) and its homocysteine analog S-(1,2-dichlorovinyl)-L-homocysteine (DCVHC) in LLC-PK1 cells. After 24-hr treatment with DCVC, the average [Ca2+]i increased from 88 +/- 23 nM to 415 +/- 92 nM. Digital image analysis revealed that the mitochondrial region, which was stained with rhodamine-123, contained lower Ca2+ concentration ([Ca2+]) than other cell areas. This distribution was different from the higher [Ca2+] in the nuclear and mitochondrial regions observed in control cells. In DCVHC-treated cells, there was also an increase in [Ca2+]i to 355 +/- 85 nM, but the increase in [Ca2+] was greater in the mitochondrial region, compared with the rest of the cell. After 72-hr treatment with DCVC or DCVHC, the average [Ca2+]i was 410 +/- 85 nM and 340 +/- 90 nM, respectively, and blebs with markedly higher [Ca2+] (600-1000 nM) than the rest of the cell appeared in both DCVC- and DCVHC-treated cells. Moreover, in DCVC-treated cells the mitochondria could not be stained with rhodamine-123, indicating severe mitochondrial damage and loss of membrane potential. All changes described above took place in viable (propidium iodide-negative) cells. The experiments demonstrate that severe perturbations of intracellular Ca2+ distribution, particularly in the mitochondrial region, precede bleb formation and cell death in the course of development of toxicity by DCVC and DCVHC.

Animals↗
PubMed1989

Cytosolic sodium concentration regulates contractility of cardiac muscle.

The present chapter provides experimental evidence to show that intracellular Na+ concentration regulates cardiac contractility effectively by altering intracellular Ca2+ concentration via the Na-Ca exchange. This steep coupling between the Na+ and Ca2+ electrochemical gradients implies that a change in intracellular Na+ concentration is accompanied by a concomitant change in intracellular Ca2+ concentration (and, therefore, contractility). Under the physiologic conditions, each cardiac action potential alters intracellular Na+ concentration in a dynamic manner. Therefore, Na-Ca exchange can regulate cardiac contraction from a beat-to-beat basis.

Animals↗
PubMed1988

Stimulation of muscarinic receptors raises free intracellular Ca2+ concentration in rat ventricular myocytes.

The effect of carbachol on free intracellular calcium concentration, ([Ca2+]i) and on intracellular hydrogen concentration (pHi) was determined from fluorescence signals obtained from rat ventricular myocytes. Application of carbachol (300 mumol/l) to quin2-loaded myocytes bathed in 2 mmol/l Ca2+-containing solution caused [Ca2+]i to increase within 7-10 minutes from 182 +/- 9 to 212 +/- 11 nmol/l (n = 4). Carbachol acted via stimulation of muscarinic receptors because atropine (1 mumol/l) either prevented or abolished the increase in [Ca2+]i. Carbachol also produced a positive inotropic effect in rat papillary muscles contracting isometrically at a frequency of 0.5 Hz and enhanced contracture in resting preparations in the presence of high extracellular Ca2+ concentration ([Ca2+]o) (20 mmol/l). The effect of carbachol on [Ca2+]i was dependent on [Ca2+]o. In the presence of 10 mmol/l [Ca2+]o, the increase in [Ca2+]i was about two times that elicited by carbachol when bath [Ca2+]o was 2 mmol/l. Reduction of [Ca2+]o to 50 mumol/l abolished the carbachol effect but did not prevent caffeine-induced Ca2+ release. The carbachol-induced rise in [Ca2+]i remained unchanged in the presence of either 10 mmol/l caffeine or 1 mumol/l ryanodine. In the absence of extracellular Na+ concentration [( Na+]o), carbachol no longer produced an increase in [Ca2+]i of cardiomyocytes and failed to enhance Na+-withdrawal contracture of the rat papillary muscle. In contrast to the effect on [Ca2+]i, carbachol did not produce any change in pHi as determined from fluorescence signals obtained from rat ventricular myocytes loaded with 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Dihydropyridine modulators of voltage-sensitive Ca2+ channels specifically regulate prolactin production by GH4C1 pituitary tumor cells.

To determine whether hormone synthesis by the GH4C1 pituitary cell line could be regulated by specifically modulating the movement of Ca2+ through voltage-sensitive channels, we have compared the effects of the dihydropyridine Ca2+ channel agonist BAY K8644 and the antagonist nimodipine on hormone production and Ca2+ current in these cells. BAY K8644 elicited, after a 10-15-h lag, a dose-dependent increase in prolactin (PRL) production as determined by measurements of total intracellular and secreted hormone. Over a 72-h period, GH4C1 cells incubated with 300 nM BAY K8644 produced 2-3 times as much total PRL as control cells. The effect on PRL was specific, since BAY K8644 did not increase growth hormone production, cell growth rate, or total cell protein. Exposing GH4C1 cells to BAY K8644 for short periods, up to 90 min, did not induce the delayed increase in PRL production observed with longer incubations. The effects of nimodipine were opposite to those of the Ca2+ channel agonist. PRL production was reduced 85% during 48-h treatment with 200 nM nimodipine, whereas growth hormone production was decreased less than 15%, and cell growth and total protein were unaffected. The actions of these two drugs on PRL production were well correlated with their effects on GH4C1 Ca2+ currents as measured by whole-cell patch-clamp recordings. BAY K8644 enhanced the magnitude of the peak Ca2+ current and shifted the current-voltage relationship such that Ca2+ channels were activated at less depolarized potentials. Nimodipine potently inhibited Ca2+ movement through the non-inactivating channel, while it antagonized the increases elicited by BAY K8644. These results indicate that PRL synthesis by GH4C1 cells can be specifically regulated by agents that enhance or block the movement of Ca2+ through voltage-sensitive channels. They also suggest that hormone synthesis by a secretory cell may be coupled to electrical activity by the opening of Ca2+ channels.

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

Effects of forskolin on intracellular sodium activity in resting and stimulated cardiac Purkinje fibers from sheep.

In the present investigation, the effects of forskolin on intracellular sodium activity were studied in quiescent and electrically stimulated cardiac Purkinje fibers from sheep using Na+-sensitive microelectrodes. Also assessed, were the effects of this promoter of cytosolic cAMP production on resting membrane potential, action potential and twitch tension. In the quiescent fibers, forskolin (12 microM) caused intracellular sodium activity to decrease in the face of cellular depolarization. This cellular depolarization was occasionally accompanied by spontaneous firing of action potentials. In the stimulated fibers, forskolin (10 microM) also caused intracellular sodium activity to decrease. Moreover, it caused a marked acceleration of phase 4 pacemaker depolarization, an elevation of the plateau of the action potential and an increase in twitch tension. When the Na+ pump was inhibited by either strophanthidin (1 microM) or by 0 mM extracellular K+, forskolin had no effect on intracellular sodium activity. In summary, the results of the present study indicate that forskolin, presumably by increasing intracellular cAMP, causes the following to occur in cardiac Purkinje fibers from sheep: (a) a decrease in intracellular sodium activity when the Na+ pump is functioning normally; (b) a promotion of membrane depolarization in quiescent fibers; (c) an increase in the steepness of the pacemaker potential in electrically stimulated fibers, and (d) an increase in the force of contraction. Therefore, forskolin will be a useful tool for investigating the role of cAMP in physiological function of cardiac cells.

Action Potentials↗
PubMed1987

Pituitary Ca2+ channels: blockade by conventional and novel Ca2+ antagonists.

We have identified several new agents that block Ca2+ channels in the rat pituitary GH4C1 cell line. These drugs, which include the diphenylbutylpiperidine antipsychotic pimozide, the calmodulin antagonist calmidazolium, and the steroidal Na+ channel toxin veratridine, were compared with several conventional Ca2+ antagonist in 45Ca2+ uptake, prolactin secretion, and whole cell patch voltage-clamp experiments. Pimozide, the most potent of these novel Ca2+ antagonists, inhibited depolarization-dependent 45Ca2+ uptake and prolactin secretion half maximally at a concentration of 100 nM, whereas calmidazolium and veratridine produced 50% inhibition at concentrations of 500 nM and 1 microM. In comparison, the three organic Ca2+ antagonists nitrendipine, verapamil, and diltiazem blocked 45Ca2+ uptake half maximally at concentrations of 2.5 nM, 1 microM, and 2.5 microM, respectively. All of the antagonists inhibited Ca2+ uptake and prolactin secretion stimulated by the dihydropyridine Ca2+ agonist BAY-K 8644 less potently than KCl-stimulated responses. In patch-clamp experiments, pimozide, veratridine, and nitrendipine blocked Ca2+ current through the slowly inactivating Ca2+ channels of GH4C1 cells. These results demonstrate that Ca2+ channels in an endocrine cell line can be blocked by a variety of molecules including sodium channel toxins and calmodulin antagonists. The data extend the pharmacological similarity between Ca2+ channels in pituitary and other excitable cells and suggest a structural similarity among several cellular proteins.

Animals↗
PubMed1987

Voltage-dependent effects of isoproterenol on cytosolic Ca concentration in rat heart.

The effect of the beta-adrenoceptor agonist, isoproterenol, on cytosolic calcium concentration ([Ca2+]i) was studied with the Ca2+-sensitive fluorescent indicator quin 2 in enzymatically dissociated rat ventricular myocytes. Under conditions in which cells have normal polarized resting membrane potential, isoproterenol (1 microM) produced a decrease in [Ca2+]i. In contrast, in the depolarized cells (by raising extracellular K+ concentration to 50 mM), isoproterenol (1 microM) caused an increase in [Ca2+]i. This isoproterenol-induced increase in [Ca2+]i in depolarized cells could be reversed by prior exposure of the cells to the Ca2+ channel blocker, verapamil (5 microM). The results indicate that isoproterenol can either decrease or increase [Ca2+]i depending on membrane potential. The actual effect of isoproterenol on [Ca2+]i at any given membrane potential probably reflects the relative contributions of isoproterenol-induced stimulation of Ca2+ buffering or effluxing activities (which favor a decrease in [Ca2+]i) and enhancement of Ca2+ influx through voltage-sensitive Ca2+ channels (which favors an increase in [Ca2+]i).

Aminoquinolines↗
PubMed1986

Na+-Ca2+ exchange contributes to increase of cytosolic Ca2+ concentration during depolarization in heart muscle.

The possible role of Na+-Ca2+ exchange in contributing to depolarization-induced increase in cytosolic Ca2+ concentration ([Ca2+]i) of isolated rat ventricular myocytes was investigated. Measured with the Ca2+-sensitive indicator quin 2, [Ca2+]i increased from 177 +/- 12 (mean +/- SE, n = 11) to 468 +/- 41 nM when cells were depolarized with solutions containing 50 mM KCl [high extracellular K+ concentration ([K+]o)]. Approximately 73% of this high-[K+]o-induced increase in [Ca2+]i was abolished by the Ca2+ channel blocker verapamil (5 microM). For cells pretreated with 10 mM caffeine to deplete the Ca2+ stored in sarcoplasmic reticulum, 50 mM KCl still produced an increase in [Ca2+]i, even in the presence of 5 microM verapamil. However, if extracellular Na+ was replaced by Li+ or tris(hydroxymethyl)aminomethane, this increase was completely abolished. The results suggest that, in addition to voltage-sensitive Ca2+ channels, voltage-sensitive Na+-Ca2+ exchange can also contribute to the increase in [Ca2+]i on depolarization. Therefore both Ca2+ transport systems may play important roles in regulating cardiac excitation and contraction.

Animals↗
PubMed1985

Lidocaine's negative inotropic and antiarrhythmic actions. Dependence on shortening of action potential duration and reduction of intracellular sodium activity.

The mechanisms by which lidocaine brings about negative inotropic effects and antiarrhythmic actions in the heart have been examined. Using sheep cardiac Purkinje fibers, we studied the effects of "therapeutic" concentrations of lidocaine (20 microM lidocaine = 5.4 micrograms/ml) on electrical activity, intracellular sodium activity, and tension. For the preparation with a normal, physiological level of intracellular sodium activity (5-9 mM), the application of lidocaine leads to the following actions: action potential duration is decreased, intracellular sodium activity falls, and twitch tension is reduced. If intracellular sodium activity is first elevated (e.g., by sodium pump inhibition) so that arrhythmogenic transient depolarizations (and the underlying transient inward current are seen) then lidocaine has the following actions: The action potential duration is reduced. There is a reduction of the magnitude of the arrhythmogenic transient depolarization (or the underlying membrane current transient inward current). The magnitude of the aftercontraction that accompanies the transient depolarization (or transient inward current) is reduced. If, after sodium pump inhibition, intracellular sodium activity is still rising-then, the application of lidocaine leads to a reduction of the rate of rise of intracellular sodium. From these results, parallel voltage-clamp experiments, and recent work by others, we conclude that lidocaine leads to the reduction of the arrhythmogenic transient depolarization and a reduction of twitch tension by decreasing the inward sodium current. These actions are mediated by a reduction in action potential duration and a reduction of intracellular sodium activity and of intracellular calcium activity (by the sodium-calcium exchange mechanism).

Action Potentials↗
PubMed1984

Effects of tonicity on tension and intracellular sodium and calcium activities in sheep heart.

We have measured the effects of changing tonicity of the bathing solution on intracellular sodium and calcium activities and tension of sheep cardiac Purkinje strands and ventricular muscle. For Purkinje strands in solutions of normal tonicity, resting membrane potential was -77.4 +/- 0.4 mV (mean +/- SE), sodium activity was 7.9 +/- 0.4 mM, and calcium activity was 98 +/- 9 nM. For ventricular muscle in solutions of normal tonicity, resting membrane potential was -86.4 +/- 1.2 mV, sodium activity was 6.9 +/- 0.5 mM, and calcium activity was 70 +/- 4 nM. Reduction of tonicity to 75% of normal in both tissues produced depolarization of a few millivolts, and sodium activity fell almost to the level predicted for simple osmotic dilution. In Purkinje strands, calcium activity fell much more than that predicted for simple osmotic dilution. Twitch contraction was reduced in the hypotonic solution. Increase of tonicity to 150% and 200% caused the resting membrane potential to become more negative. In both tissues, sodium activity increased somewhat less than predicted from simple water movement, and calcium activity increased proportionately much more than sodium activity. The much larger change of calcium activity in both hypo- and hypertonic solutions could be explained by water movement plus the effect of sodium-calcium exchange. In hypertonic solutions, tonic tension was increased, along with the rise in calcium activity; however, the twitch tension was reduced. This reduction of twitch tension may be due to a direct effect of hypertonicity on cross-bridge behavior, as has been reported for skeletal muscle.

Animals↗
PubMed1984

Measurement of cytosolic free calcium concentration in isolated rat ventricular myocytes with quin 2.

Cytosolic free calcium concentration was determined in isolated ventricular myocytes from adult rats with the calcium-sensitive indicator, quin 2. The fluorescence signal from resting cells indicated that cytosolic free calcium concentration was 181 +/- 18 nM (mean +/- SEM, n = 18). Inhibition of the sodium-potassium pump with strophanthidin (0.1 mM) resulted in an increase of cytosolic free calcium concentration from 186 +/- 17 to 736 +/- 129 nM (n = 6). The results indicate that it is possible to measure cytosolic free calcium concentration in cardiac muscle cells that have been isolated enzymatically. Moreover, they confirm the observation that inhibition of the sodium-potassium pump increases cytosolic free calcium concentration, presumably via the sodium-calcium exchange mechanism.

Aminoquinolines↗
PubMed1983

The role of intracellular sodium activity in the anti-arrhythmic action of local anaesthetics in sheep Purkinje fibres.

The effects of lidocaine have been examined on the arrhythmogenic transient inward current (ITI) in voltage-clamped sheep cardiac Purkinje fibres. Tension and intracellular Na activity (aiNa) were measured simultaneously. The addition of lidocaine (200-300 microM) produced an immediate decrease of inward holding current and a gradual fall of aiNa. The relative magnitudes of the changes of current and aiNa were shown to be consistent with the outward shift of current representing principally a reduction of inward Na current. The Na pump was inhibited by reducing the external Rb concentration in a K-free solution. This produced an after-contraction and transient inward current (ITI) along with a rise of aiNa. The subsequent addition of lidocaine decreased the magnitude of ITI and the after-contraction while decreasing aiNa. Tetrodotoxin (TTX) had qualitatively similar effects to lidocaine on inward holding current, aiNa, ITI and the after-contraction. When aiNa was changed by (i) lidocaine, (ii) TTX or (iii) small changes of external Rb concentration, a hysteresis was seen in the relationship between aiNa and ITI or after-contraction. The hysteresis was similar to that previously found between aiNa and contraction (Eisner, Lederer & Vaughan-Jones, 1981). Despite this hysteresis, neither lidocaine nor TTX affected the relationship between magnitudes of ITI and the after-contraction. It is suggested that the fall of aiNa is a major factor in the reduction of ITI by lidocaine. These results are discussed in relation to the anti-arrhythmic actions of lidocaine.

Action Potentials↗
PubMed1983

Effects of calcium on membrane potential and sodium influx in barnacle muscle fibers.

The influence of internal and external Ca2+ on membrane potential and 22Na influx were tested in internally perfused giant barnacle muscle fibers. The fibers depolarized by about 2-3 mV, and Na+ influx increased when external Ca2+ was removed. These effects were inhibited and reversed by adding 2 mM La3+ externally but not by tetrodotoxin (TTX). Ca2+ channel blockers did not prevent the depolarization. Increasing internal free Ca2+ ([Ca2+]i) from 10(-7) to 10(-5) M also stimulated Na+ influx and depolarized the fibers by a few millivolts. Neither external La3+ nor TTX prevented the effects of raising [Ca2+]i; however, internal tetrabutylammonium ions depolarized the fibers and attenuated the internal Ca2+-dependent effects. These data are consistent with the idea that removal of external Ca2+ activates a La3+-sensitive channel that is permeable to Na+; raising [Ca2+]i activates a La2+-insensitive, Na+-permeable channel that may be similar to the internal Ca2+-activated nonselective cation channels observed in cardiac muscle. The results demonstrate that all Na+ (and Ca2+) fluxes that do not contribute to Na-Ca exchange must be carefully identified before the exchange stoichiometry can be determined from Na+ and Ca2+ flux measurements.

Animals↗