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D A Eisner

Publications and source records attributed to D A Eisner.

At least 109 records · Page 6Linked to original sources
PubMed1987

The contribution of intracellular acidosis to the decline of developed pressure in ferret hearts exposed to cyanide.

1. The concentrations of phosphorus-containing metabolites were measured using 31P nuclear magnetic resonance (n.m.r.) in Langendorff-perfused ferret hearts. The hearts were stimulated at a constant rate and developed pressure was measured. 2. The inhibition of oxidative phosphorylation with cyanide (2 mM) decreased developed pressure. This was accompanied by an intracellular acidosis, a fall in the concentration of phosphocreatine ([PCr]) and a rise in that of inorganic phosphate ([Pi]). 3. The effects of cyanide on developed pressure were compared with those of simply decreasing the intracellular pH (pHi) (by elevating CO2) to match the change produced by cyanide. The change of pHi alone resulted in about 33% of the decrease of force produced by cyanide. 4. The relationship between developed pressure and pHi was investigated by altering CO2. It could be described by an equation of the form:log developed pressure = a + b pHi where a and b were independent of pHi. The addition of cyanide decreased a but had no significant effect on b. In other words a given change of pHi had the same fractional effect on pressure in the absence as in the presence of cyanide. 5. In another series of experiments, after cyanide had been added, pHi was returned to control levels by decreasing CO2. This increased developed pressure. Nevertheless the pressure was still considerably less than in control. Furthermore, if the acidosis was abolished by decreasing CO2 at the same time as cyanide was added developed pressure still decreased. 6. We conclude that most of the decrease of developed pressure produced by cyanide is not produced by intracellular acidosis and may result from increased [Pi].

Animals↗
PubMed1986

The role of intracellular Ca ions in the therapeutic and toxic effects of cardiac glycosides and catecholamines.

Many inotropic maneuvers act by increasing the intracellular calcium concentration [( Ca2+]i). The present report illustrates this with respect to the positive inotropic effects of cardiac glycosides and catecholamines. It is shown that the increased contractility produced by cardiac glycosides is accompanied by an increase in intracellular Na concentration and, furthermore, that the relationship between contraction and Na is very steep. This steep dependence, which may result from a Na-Ca exchange which exchanges several Na ions per Ca, means that maneuvers that have only small effects on Na will have significant effects on contraction. Cardiac glycosides also produce abnormal pacemaker activity and cardiac arrhythmias. These originate from a transient inward current activated by oscillations of [Ca2+]i, which result from spontaneous oscillatory release of Ca ions from the sarcoplasmic reticulum. The local anesthetic group of antiarrhythmic agents abolishes the transient inward current. Catecholamines also increase systolic [Ca2+]i and, in high enough concentrations, can produce oscillations of [Ca2+]i. This tendency of glycosides and catecholamines to produce arrhythmogenic oscillations of [Ca2+]i is a major limitation to their use. Therefore, inotropic agents that act by means other than increasing [Ca2+]i may be of great efficacy.

Animals↗
PubMed1986

A study of intracellular calcium oscillations in sheep cardiac Purkinje fibres measured at the single cell level.

Previous work has shown that an elevation of intracellular calcium concentration [( Ca2+]i) produces spontaneous oscillations of [Ca2+]i. However the fact that the oscillations are unsynchronized between different cells has made it difficult to study them. We have therefore injected only one cell in a Purkinje fibre with aequorin in order to avoid these problems. The addition of strophanthidin (10 microM) produced an increase of mean aequorin light over the course of several minutes. During this period spontaneous oscillations of light developed and, with time, their frequency and magnitude increased. The oscillations could first be seen at levels of [Ca2+]i of less than 1 microM. The amplitude of the oscillations of [Ca2+]i could be up to 10 microM and was modulated at a slow rate (about 0.3-0.5 Hz). This suggests that, even within one cell, different regions may oscillate at different frequencies. Elevating [Ca+]o, removing extracellular Na+, or depolarization increased the magnitude of the aequorin light oscillations. Converting the records to [Ca2+]i showed that this increase in the magnitude of the aequorin oscillations was accompanied by a real increase of mean [Ca2+]i and of the magnitude of the oscillations [Ca2+]i. The frequency of the oscillations increased up to a point but saturated at a maximum value of 3-4 Hz. Since previous experiments have used the mean aequorin light to estimate mean [Ca2+]i, we have calculated the error produced in this calculation by the presence of [Ca2+]i oscillations. We estimate that the error is greatest at low levels of Ca2+ loading when the frequency of the oscillations is low. However, at higher Ca2+ loads, when the frequency is above 2 Hz, the error is probably less than 10%. If oscillations were produced by removal of external Na+ after the application of strophanthidin, then either ryanodine or caffeine abolished the oscillations. Furthermore, in both cases, the resulting steady level of [Ca2+]i was similar to the mean level before the addition of the drugs. In another series of experiments we examined the effects of these drugs on oscillations produced by the application of strophanthidin. Caffeine produced a transient increase in both the frequency of the oscillations and mean [Ca2+]i before abolishing the oscillations and decreasing [Ca2+]i to below the level in the absence of caffeine. In contrast ryanodine gradually decreased both the mean [Ca2+]i and the frequency until the oscillations were abolished. During this period of slowing of the oscillations their magnitude was often increased.

Aequorin↗
PubMed1986

Measurement of intracellular calcium during the development and relaxation of tonic tension in sheep Purkinje fibres.

The photoprotein aequorin was micro-injected into several cells in a sheep Purkinje fibre. The intracellular Ca concentration [( Ca2+]i) was measured from the resulting light emission. Inhibition of the Na-K pump with strophanthidin resulted in the development of tonic tension which increased on depolarization. This increase was accompanied by an increase of aequorin light. Increasing external Ca concentration [( Ca2+]o) or the magnitude of the depolarization increased both light and tension. If the depolarizing pulse was maintained for several minutes then both tonic tension and aequorin light slowly decayed. The relationship between tension and light was unaffected during this decay. On repolarization the light decayed to below the level before the depolarization before slowly increasing. During this period a test depolarization produced increases of aequorin light and tension which were smaller than control. The application of ryanodine (1-10 microM) abolished all components of tension other than the tonic component. Under these conditions the time course of the increase of tonic tension and aequorin light on depolarization was sufficiently slow to be measured. In most (five out of six) experiments the relationship between light and tension during this development of tonic tension was found to be similar to that during the subsequent spontaneous decay. However, in one experiment the decay of force was greater than could be accounted for by the fall of [Ca2+]i. It is concluded that most of the spontaneous relaxation of tonic tension can be attributed to a fall of [Ca2+]i rather than to other explanations such as an intracellular acidification or increase of inorganic phosphate concentration.

Aequorin↗
PubMed1986

Metabolic consequences of increasing intracellular calcium and force production in perfused ferret hearts.

31P nuclear magnetic resonance was used to measure the relative concentrations of phosphorus-containing metabolites in Langendorff-perfused ferret hearts. Intracellular concentrations of inorganic phosphate ([Pi]i), phosphocreatine [( PCr]i), ATP ([ATP]i) and [H+] (pHi) were determined. Exposure of the heart to strophanthidin (10-40 microM) produced an increase in developed pressure over 5-10 min. In the presence of strophanthidin, [ATP]i was unchanged, [PCr]i showed a small fall, [Pi]i showed a small rise and there was a small acidosis. Exposure of the heart to a solution in which all the Na had been replaced by K (0 Na(K) solution) produced an increase of resting pressure which then decayed. During this contracture [PCr]i fell transiently and [Pi]i rose transiently with approximately the same time course as the contracture. However, [ATP]i remained constant throughout. The exposure to the 0 Na(K) solution also produced an intracellular acidosis. The changes in [PCr]i and [Pi]i and the intracellular acidosis were all increased during perfusion with 0 Na(K) if the heart had previously been exposed to strophanthidin. The efflux of lactate from the heart was increased during the exposure to the 0 Na(K) solution. The magnitude of this increase was enhanced by prior exposure to strophanthidin. The increase of intracellular lactate (calculated from this efflux) was sufficient to account for the observed intracellular acidification. An increase of lactate efflux could also be measured when an isolated papillary muscle was exposed to the 0 Na(K) solution. The intracellular acidification produced by Na removal was substantially decreased after prevention of glycolysis either by substrate depletion or by the application of iodoacetate. Elevation of the extracellular calcium concentration ([Ca2+]o) produced a large increase of developed pressure which was accompanied by a small transient increase of [Pi]i, a decrease of [PCr]i and a small intracellular acidosis. There was also an increase of lactate efflux. After exposure to strophanthidin the same increase of [Ca2+]o decreased developed pressure. The associated rise in [Pi]i and fall in [PCr]i were increased but there was no significant acidosis under these conditions. In addition to other explanations (Allen, Eisner, Pirolo & Smith, 1985a), it is likely that the fall of force is partly accounted for by the rise of [Pi]i.

Adenosine Triphosphate↗
PubMed1986

Effects of membrane potential on intracellular calcium concentration in sheep Purkinje fibres in sodium-free solutions.

1. The intracellular Ca2+ concentration [( Ca2+]i) was measured in voltage-clamped sheep cardiac Purkinje fibers while recording tension simultaneously. 2. When [Na+]i was elevated (by Na+-K+ pump inhibition) depolarization produced an increase of tonic tension. 3. Replacement of external Na+ by Li+ or choline produced a contracture which then relaxed spontaneously. Following this relaxation, depolarization either had no effect on tonic tension or produced a small decrease. 4. When external Na+ was replaced by Ca2+, depolarization (over the range -120 to -20 mV) produced a decrease of tonic tension and [Ca2+]i. Hyperpolarization increased tonic tension and [Ca2+]i. 5. An after-contraction and accompanying increase of [Ca2+]i were produced by repolarization in both Na+-free and Na+-containing solution. This eliminates the possibility that the stimulus for the after-contraction is the increase of [Ca2+]i during the depolarization and suggests that the stimulus may be the change of membrane potential. 6. The increase of [Ca2+]i on hyperpolarization seen in Na+-free solutions persisted in the presence of ryanodine. 7. These results show, in contrast to previous work, that in Na+-free solutions tonic tension is still sensitive to membrane potential. The results support the hypothesis that, in Na+-containing solutions, the increase of tonic tension on depolarization results from a voltage-dependent Na+-Ca2+ exchange. The reduction of tonic tension on depolarization in Na+-free solutions may be due to the decrease of the electrochemical gradient for Ca2+ to enter the cell.

Aequorin↗
PubMed1985

The effects of intracellular Na on contraction and intracellular pH in mammalian cardiac muscle.

Intracellular Na and pH were measured with recessed-tip ion-selective microelectrodes in voltage-clamped sheep cardiac Purkinje fibers. Intracellular Na activity (aiNa) was elevated by inhibiting the Na/K pump. This produced an increase of twitch tension that had a steep dependence on the increase of aiNa. These effects of aiNa on twitch tension are probably mediated by an Na-Ca exchange. An increase of aiNa also produced a component of tonic tension that appears to be produced directly by the Na-Ca exchange. The dependence of tonic tension and aiNa on membrane potential suggests that this exchange process may be voltage-sensitive. The increase of aiNa is associated with an intracellular acidification that appears to be secondary to an increase of [Ca2+]i produced by Na-Ca exchange. Therefore, as well as affecting [Ca2+]i, Na-Ca exchange can under some circumstances influence pHi indirectly, and this complicates the interpretation of changes in tension, since protons and Ca ions have opposite effects on contractile force.

Animals↗
PubMed1985

The influence of chemical agents on the level of ionized [Ca2+] in squid axons.

Squid giant axons injected with either aequorin or arsenazo III and bathed in 3 mM Ca (Na) seawater were transferred to 3 mM Ca (K) seawater and the response of the aequorin light or the change in the absorbance of arsenazo III was followed. These experimental conditions were chosen because they measure the change in the rate of Na/Ca exchange in introducing Ca into the axon upon depolarization; [Ca]o is too low to effect a channel-based system of Ca entry. This procedure was applied to axons treated with a variety of compounds that have been implicated as inhibitors of Na/Ca exchange. The result obtained was that the substances tested could be placed in three groups. (a) Substances that were without effect on Ca entry effected by Na/Ca exchange were: D600 at 10-100 microM, nitrendipine at 1-5 microM, Ba2+ and Mg2+ at concentrations of 10-50 mM, lidocaine at 0.1-10 mM, cyanide at 2 mM, adriamycin at a concentration of 3 microM, chloradenosine at 35 microM, 2,4-diaminopyridine at 1 mM, Cs+ at 45-90 mM, and tetrodotoxin at 10(-7). (b) Substances that had a significant inhibitory effect on Na/Ca exchange were: Mn2+, Cd2+, and La3+ at 1-50 mM, and quinidine at 50 microM. (c) There were also blocking agents and biochemical inhibitors whose action appeared to be the inhibition of nonmitochondrial Ca buffering in axoplasm rather than an inhibition of Na/Ca exchange. These were the general anesthetic l-octanol at 0.1 mM and 1 mM orthovanadate plus apyrase.

Aequorin↗
PubMed1985

Effects of caffeine, tetracaine, and ryanodine on calcium-dependent oscillations in sheep cardiac Purkinje fibers.

Membrane current and tension were measured in voltage-clamped sheep cardiac Purkinje fibers. Elevating the intracellular calcium concentration ([Ca2+]i) results in oscillations of membrane current and tension both at rest and during stimulation. During stimulation, an oscillatory transient inward current and an after contraction follow repolarization. We have examined the effects on the oscillations of changing the extracellular calcium concentration ([Ca2+]o) and of adding various drugs. In agreement with previous work, high concentrations of drugs that affect the sarcoplasmic reticulum, namely caffeine (10-20 mM), tetracaine (1 mM), and ryanodine (10 microM), abolish the oscillations. However, at lower concentrations, these three drugs have different effects on the oscillations. Caffeine (1-2 mM) decreases the oscillation amplitude but increases the frequency. Tetracaine (100-500 microM) has little effect on the magnitude of the oscillations but decreases their frequency. Ryanodine, at all concentrations used (0.1-10 microM), eventually abolishes the oscillations but, in doing so, decreases the magnitude, leaving the frequency unaffected. When [Ca2+]o was changed in order to vary [Ca2+]i, both the frequency and the magnitude of the oscillations always changed in the same direction. This suggests that these three drugs have effects in addition to just changing [Ca2+]i.

Alkaloids↗
PubMed1985

The relationship between intracellular calcium and contraction in calcium-overloaded ferret papillary muscles.

The photoprotein aequorin was microinjected into cells on the surface of ferret papillary muscles. Tension and aequorin light, a function of intracellular Ca concentration ([Ca2+]i), were monitored. The preparations were exposed to increased concentrations of extracellular Ca ([Ca2+]o). Small increases in [Ca2+]o led to the usual increase in both the systolic light signal and the developed tension. However, high [Ca2+]o led to a fall in developed tension known as Ca overload. This fall of tension was seen at lower [Ca2+]o if the experiment was performed in the presence of strophanthidin. In conditions of Ca overload, the systolic light signal was either similar in amplitude or larger than the systolic light observed under conditions which did not lead to Ca overload. Oscillations of diastolic light were invariably present under conditions of Ca overload. These oscillations were accompanied by after-contractions which were small in relation to the magnitude of the aequorin light oscillations. During Ca overload, the variance of the amplitude of the systolic light signal was greater than could be accounted for by the random nature of the arrival of photons. Small systolic light signals occurred when there had been an oscillation of light in the diastolic period immediately preceding the systolic light signal. Large systolic light signals occurred when the preceding period (approximately 1 s) was free of oscillations of light. These observations suggest that if the sarcoplasmic reticulum (s.r.) has spontaneously released its contents of Ca as shown by a diastolic [Ca2+]i oscillation, then a stimulated systolic Ca signal occurring within about the next second is smaller, possibly because it takes longer than this period for the s.r. to reload with Ca. If this process occurs randomly in the various cells of the preparation, developed tension will be reduced because those cells which have a small Ca release will act as a compliance in series with cells which have a large Ca release.

Aequorin↗
PubMed1985

The mechanism of the increase of tonic tension produced by caffeine in sheep cardiac Purkinje fibres.

The effects of caffeine were examined on contraction and membrane current in voltage-clamped sheep cardiac Purkinje fibres. The photoprotein aequorin was injected into several cells in order to measure the intracellular ionized Ca concentration [( Ca2+]i). When the Na-K pump was inhibited, depolarization produced a twitch followed by a tonic component of tension. Repolarization produced an after-contraction. These components of tension were accompanied by corresponding increases of aequorin light. Caffeine (10 mM) decreased both the twitch and the after-contraction while increasing the tonic component. The application of caffeine also produced a transient increase of aequorin light, both during depolarization and at rest, which was followed by a maintained decrease in all three components of the light signal. In particular, although caffeine decreased the rise of aequorin light during prolonged depolarization it increased the tonic tension. The possibility that the effects of caffeine on tonic tension could be due to suppression of spontaneous Ca oscillations was rejected for the following reasons. (i) Ryanodine (which also abolishes Ca oscillations) decreased the magnitude of the tonic tension. (ii) Caffeine still increased tonic tension when it was added to a fibre exposed to ryanodine (1-10 microM). In the presence of ryanodine it was possible to measure [Ca2+]i and tonic tension without interference from Ca oscillations. The increase of tonic tension produced by caffeine could not be accounted for by a rise of [Ca2+]i. The results showed that, at a given level of Ca, caffeine increased tension. The results show that a large part of the increase of tonic tension produced by caffeine is due to an increase of the Ca sensitivity of the contractile apparatus rather than to changes of [Ca2+]i. The consequence of this observation for the experimental use of caffeine is discussed.

Action Potentials↗
PubMed1985

Na-Ca exchange: stoichiometry and electrogenicity.

This review discusses the evidence concerning the stoichiometry of Na-Ca exchange. In particular we consider whether the Na-Ca exchange has been shown to transport more than two Na+ ions per Ca2+ ion and therefore whether it generates an electric current. The first part of this review discusses both direct and indirect evidence concerning the stoichiometry of the exchange and its possible voltage dependence. We find that, although there is some evidence suggesting that more than two Na+ ions may exchange for each Ca2+ ion, most of the available evidence is equivocal and cannot fix the stoichiometry precisely. Furthermore, using a simple and explicit circulating carrier model for the Na-Ca exchange, we show that the effect of membrane potential on the Na-Ca exchange may be considerably more complicated than is generally believed. In particular we find that both electrogenic and electroneutral exchanges will be affected by membrane potential. We therefore conclude that the demonstration of the voltage dependence of the Na-Ca exchange does not necessarily imply that it is electrogenic. Additionally, this analysis shows that, apart from a restricted range near thermodynamic equilibrium, it is impossible to predict either the magnitude or the direction of the effects of membrane potential on the exchange. In the second part of the review we consider whether any known membrane currents may be attributed to Na-Ca exchange. We show, in contrast to previous suggestions, that the Na-Ca exchange can theoretically produce a current that appears to be activated by intracellular Ca and that has a reversal potential. However, the experimental demonstration that a given current is produced by Na-Ca exchange is hampered by the existence of other Ca- and Na-dependent currents. In conclusion, we feel that there is no evidence that allows any particular membrane current to be unambiguously identified with the Na-Ca exchange.

Action Potentials↗
PubMed1985

The effects of ryanodine on calcium-overloaded sheep cardiac Purkinje fibers.

Prolonged exposure to high concentrations of strophanthidin produces an initial increase followed by a subsequent decrease of twitch tension. The slow decrease is termed calcium overload. The aim of the present work was to investigate the effects of ryanodine (an inhibitor of calcium release from the sarcoplasmic reticulum) on calcium-overloaded sheep cardiac Purkinje fibers. The fibers were voltage-clamped, and tension was measured while monitoring the intracellular calcium concentration with the photoprotein aequorin. When strophanthidin (10 microM) was applied to produce calcium overload, a depolarizing pulse produced twitch, and tonic components of tension and repolarization produced an aftercontraction. These components of tension were accompanied by corresponding increases of aequorin light. Ryanodine (1 microM) gave a transient increase of twitch tension. The twitch then decreased to very low levels. The aftercontraction and its corresponding aequorin light signal decreased monotonically on application of ryanodine. It has been suggested that the fall of force in calcium overload may be due to random diastolic release of calcium from the sarcoplasmic reticulum interfering with subsequent systolic calcium release. We suggest that the positive inotropic effect of ryanodine can be explained if ryanodine decreases the diastolic release of calcium. The transient positive inotropic effect of ryanodine reported here is therefore consistent with the hypothesis that the fall of force in calcium overload is due to diastolic calcium oscillations.

Alkaloids↗
PubMed1984

A comparison of measurements of intracellular Ca by Ca electrode and optical indicators.

Squid giant axons were injected with aequorin or arsenazo III and impaled with a Ca-sensing electrode. The light output of aequorin or the spectrophotometer output when measuring arsenazo was compared with the voltage output of the electrode when the squid axon was depolarized with high-K solutions, when the seawater was made Na-free, or when the axon was tetanized for several minutes. The results from these treatments were that the optical response rose (as much as 50-fold) with all treatments known to increase Ca entry, while the electrode remained unaffected by these treatments. If axons previously subjected to Ca load are treated with electron-transport poisons such as CN, it is known that [Ca]i rises after a time necessary to deplete ATP stores. In such axons one expects a rise of [Ca]i in axoplasm which does not necessarily have to be uniform although the source of such Ca is the mitochondria and these are uniformly distributed in axoplasm. Under conditions of CN application, the optical signals from aequorin or arsenazo and Ca electrode output do rise together when [Ca]i is high, but there is a region of [Ca]i concentration where aequorin light output or arsenazo absorbance rises while electrode output does not. Axons not loaded with Ca but injected with apyrase and vanadate have mitochondria that still retain some Ca and this can be released by CN in a truly uniform manner. The results show that such a release (which is small) can be readily measured with aequorin, but again the Ca electrode is insensitive to such [Ca]i change.

Aequorin↗
PubMed1984

Control of intracellular ionized calcium concentration by sarcolemmal and intracellular mechanisms.

The regulation of the resting intracellular ionized calcium concentration [( Ca2+]i) has been studied in ferret papillary muscle using the photoprotein aequorin to measure [Ca2+]i. Elevating [Ca2+]o produced an initial rapid increase of [Ca2+]i and tension which then decayed to a steady level. This secondary fall of [Ca2+]i is attributed to a secondary decrease of Ca entry on Na-Ca exchange produced by the known fall of [Na+]i. Replacing external Na by K produced a large transient increase of both [Ca2+]i and tension which then decayed spontaneously to near the resting level. If Na was removed after metabolic inhibition with cyanide and deoxyglucose then neither tension nor [Ca2+]i recovered. The addition of the mitochondrial uncoupler FCCP to a muscle in Na-free solution produced a gradual rise of tension but only elevated [Ca2+]i after a delay of many minutes. Similarly caffeine did not elevate [Ca2+]i. These experiments do not support the hypothesis that the regulation of resting [Ca2+]i in Na-free solutions depends solely on intracellular sequestration of [Ca2+]i. The first twitch elicited in Na-containing solutions after exposure to Na-free solution was much larger than control and was associated with a large Ca transient attributed to increased loading of the sarcoplasmic reticulum with Ca in the Na-free solution. The elevation of [Ca2+]i in Na-free solutions was accompanied by spontaneous fluctuations of both [Ca2+]i and tension with a frequency of about 3 Hz. These fluctuations were abolished by drugs such as caffeine or ryanodine which interfere with sarcoplasmic reticulum function. These results provide direct evidence for the spontaneous release of Ca from the sarcoplasmic reticulum inferred from previous, less direct, work.

Animals↗
PubMed1984

Factors influencing free intracellular calcium concentration in quiescent ferret ventricular muscle.

The photoprotein aequorin was injected into cells of ferret papillary muscles to monitor the resting intracellular free Ca concentration [( Ca2+]i). Increasing the external Ca concentration [( Ca2+]o) increased both resting [Ca2+]i and resting tension. The tension and [Ca2+]i both rose to a peak and then declined to a steady-state level which was higher than the control. Qualitatively similar, but larger, effects were observed if [Ca2+]i was first elevated with strophanthidin. The increase of [Ca2+]i was accompanied by the development of spontaneous oscillations of [Ca2+]i. When a steady level of [Ca2+]i had been reached in high [Ca2+]o, [Ca2+]o was reduced back to the control level for a brief period. A subsequent increase of [Ca2+]o produced a rise of [Ca2+]i to the same steady level as that previously found in the high [Ca2+]o but the initial peak and subsequent decline were absent. It is suggested that the decline of [Ca2+]i from the initial peak is mediated by a fall of intracellular Na concentration [( Na+]i) limiting Ca entry on a Na-Ca exchange. Increasing external K concentration [( K+]o) from 5 to 30 mmol/l had no detectable effect on [Ca2+]i under control conditions. However, if [Ca2+]i was first increased either by applying strophanthidin or by increasing [Ca2+]o, increasing [K+]o produced a transient rise of [Ca2+]i and tension. This rise was unaffected by D600. It is suggested that the secondary decline of [Ca2+]i after the initial rise may, again, be produced by a fall of [Na+]i acting on an Na-Ca exchange. Acidification produced by increasing [CO2] had no detectable effect on [Ca2+]i under control conditions. However, if [Ca2+]i was increased by strophanthidin, acidification produced a rise of [Ca2+]i. This rise of [Ca2+]i was partly transient even when the intracellular acidification was presumably maintained (raising CO2 at constant [HCO3-]). Acidification in Na-free solutions had qualitatively similar effects to those in Na-containing solutions. In Na-free solutions (Na replaced by K) the [Ca2+]i could be maintained at a low level for at least several hours. Increases of [Ca2+]o in Na-free solutions led to a decrease of [Ca2+]i, and similarly decreasing [Ca2+]o led to an increase in [Ca2+]i. These anomalous effects of [Ca2+]o on [Ca2+]i could be abolished by Mn ions or D600. It is suggested that changes in [Ca2+]o may have reciprocal effects on Ca permeability and hence on [Ca2+]i. The application of the mitochondrial uncoupler FCCP in Na-free solutions led to an increase of resting tension followed, after a substantial delay, by an increase of [Ca2+]i.(ABSTRACT TRUNCATED AT 400 WORDS)

Aequorin↗