Rise In Intracellular Na Research Articles

Short-term effect of epidermal growth factor (EGF) on sodium and glucose cotransport of isolated jejunal epithelial cells

This study was undertaken to assess the short-term effects of EGF on sodium and glucose uptake, glucose metabolism and Na + K +- ATPase activity in isolated enterocytes of rats. Jejunal cells exposed to EGF had a significantly greater total uptake of sodium compared to controls after 6 min. Kinetic analysis of glucose transport across BBMV's demonstrated similar K m values but a significant increase of the V max in vesicles prepared from cells first exposed to EGF as compared to controls. EGF was also associated with a significant increase in glucose metabolism of jejunal enterocytes after 15 min. The activity of Na + K +- ATPase increased in jejunal enterocytes exposed to EGF. The increase in Na + K +- ATPase activity of the cells following EGF exposure was not accompanied by an increase in immunodetectable total or assembled Na + K +- ATPase protein. EGF's effect on enzyme activity was abolished by removing NaCl from the incubation solution, and by preincubating the enterocytes with phlorizin prior to addition of EGF. Preincubation with amiloride did not inhibit the effect of EGF on Na + K +- ATPase . The results confirm that EGF promotes uptake of both sodium and glucose by the jejunal mucosal cells, and suggest the effect of EGF on glucose and sodium is mediated through the brush-border membrane glucose-sodium transporter. The increase in Na + K +- ATPase activity that occurs with EGF appears to be secondary to a rise in intracellular Na + concentration. The short-term effects of EGF on glucose and sodium transport by the small intestine may have potential therapeutic implications.

Release of calcium from guinea pig cardiac sarcoplasmic reticulum induced by sodium-calcium exchange

Depolarisation-induced Na+ influx through tetrodotoxin sensitive Na+ channels causes a rapid increase in intracellular Ca2+ concentration ([Ca2+]i). The Na+ current (INa) induced [Ca2+]i transients: (a) occur after blocking sarcolemmal Ca2+ channels with nisoldipine or D-600, (b) are inhibited by ryanodine, and (c) are dependent upon extracellular Ca2+. Thus the INa induced [Ca2+]i transients arise from sarcoplasmic reticular Ca2+ release triggered by Ca2+ entering the myocyte, following a transient rise in intracellular Na+ ([Na+]i), via a pathway distinct from sarcolemmal Ca2+ channels. Reverse mode Na(+)-Ca2+ exchange could provide such a pathway for Ca2+ entry. The aim of this study was to ascertain directly whether Na(+)-Ca2+ exchange mediates the INa induced release of Ca2+ from sarcoplasmic reticulum. Whole cell voltage and current clamped guinea pig ventricular myocytes dialysed with indo-1 were used; Ca2+ transients elicited upon activation of INa before and after inhibiting the exchanger were measured. Following conditioning protocols to load Ca2+ stores, activation of INa during a test pulse to -50 mV from a holding potential of -80 mV elicited [Ca2+]i transients in caesium loaded myocytes superfused with solutions containing 2.5 mM Ca2+ and 5 microM nisoldipine. When extracellular Na+ was replaced with equimolar lithium, which carries current through Na+ channels but does not readily substitute for Na+ on the Na(+)-Ca2+ exchanger, or when Ni2+ (5 mM) or dichlorobenzamil (10 microM), which block the exchanger, were added to superfusion solutions, activation of INa failed to elicit [Ca2+]i transients. Lithium and Ni2+ also inhibited nisoldipine insensitive [Ca2+]i transients elicited by action potentials, indicating that INa and Na(+)-Ca2+ exchange may play a role in excitation-contraction coupling under physiological conditions. Activation of INa appears to promote Ca2+ entry into cardiac cells by stimulation of reverse mode Na(+)-Ca2+ exchange, triggering Ca2+ release from the sarcoplasmic reticulum.

Increased intracellular Na+ augments mobilization of Ca2+ from SR in vascular smooth muscle cells.

The effect of a rise in intracellular Na+ concentration ([Na+]cyt) on the amount of Ca2+ in intracellular stores was studied in vascular smooth muscle cells from the A7r5 line. The relative amount of stored Ca2+ was estimated in fura 2-loaded cells by the rise in cytosolic free Ca2+ concentration ([Ca2+]cyt) evoked by Ca2+ release from the sarcoplasmic reticulum (SR). To improve the detection of released Ca2+, extrusion of Ca2+ from the cytosol was minimized by using nominally Na+/Ca(2+)-free medium containing 0.5 mM La3+ [for vasoconstrictor experiments, the medium contained 0.5 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid and no La3+]. Ca2+ release was triggered by thapsigargin (TG), an SR Ca(2+)-ATPase inhibitor, and by the vasoconstrictors arginine vasopressin (AVP) and serotonin (5-HT). Incubation with 1-3 mM ouabain for 20 min, which raises [Na+]cyt from 4.4 to 9.0 mM, increased "resting" [Ca2+]cyt only slightly (from 87 to 122 nM). However, ouabain greatly augmented the release of Ca2+ evoked by TG [from 639 nM (control) to 1,021 nM], by AVP (from 993 to 1,597 nM), and by 5-HT (from 559 to 1,486 nM). Ouabain-induced augmentation of TG-evoked Ca2+ release was not affected by 10 microM verapamil; this implies that the effect of ouabain was not due to Ca2+ entry through voltage-gated Ca2+ channels. The response to TG was not augmented when ouabain was applied for 20 min in Na(+)-free medium (Na+ replaced by equimolar N-methyl-D-glucamine) to prevent [Na+]cyt from rising.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in K+, Na+ and calcium contents during in vivo stimulation of rat skeletal muscle.

The effects of in vivo stimulation via the sciatic nerve on Na+, K+ and calcium contents in slow-twitch and fast-twitch muscles were compared. Whereas intermittent stimulation for 24 h at 20 Hz caused only minor changes in soleus (SOL), a considerable loss of K+ (around 24%) and gain of Na+ (around 84%) was observed in extensor digitorum longus (EDL) and tibialis anterior (TA) muscles. These changes could be detected within 0.5 h and a plateau was maintained from 2 to 24 h. Total calcium content increased progressively, reaching values 245 and 382% above the control level in EDL and TA muscle, respectively, after 24 h of 20 Hz stimulation. Whereas the Na+ and K+ content recovered within a few hours, calcium content did not return towards control level until after 48 h of rest. In a pilot study performed with continuous stimulation at 10 Hz, the changes in Na+ and K+ contents in SOL, EDL and TA muscle were comparable to those at 20 Hz. The concentration of the Na(+)-K+ pumps was highest in the fast-twitch EDL and TA muscles and was unaffected by 10 Hz stimulation. It is concluded that a stimulation pattern leading to a rise in intracellular Na+ and a loss of K+ may cause a marked accumulation of calcium. These events seem to be related to insufficient activation of the Na-K+ pump rather than to variations in the total Na(+)-K+ pump capacity.

Regulation of the dynamic properties of platelet plasma membrane by intracellular sodium ions

Our previous experiments in human and rat platelets demonstrated that the absence of extracellular Na + increased the fluorescence anisotropy of TMA-DPH (trimethylamino-diphenylhexatriene, probe preferentially incorporated into the outer leaflet of the plasma membrane). Here we investigated further the in vitro effects of Na + ions on membrane dynamic properties. Na +-dependent changes were reversible and they required about 10–20 min to be induced. They were specifically located in the TMA-DPH environment because they were not observed with diphenylhexatriene (probe non-selectively incorporated into all hydrophobic domains of the cell). To evaluate the possible influence of the intracellular Na +, the effects of sodium replinishment, monensin, ouabain and thrombin on TMA-DPH anisotropy were measured. A rise in intracellular Na + above the physiological level was associated with unchanged or slightly decreased TMA-DPH anisotropy whereas its decrease was accompanied by a pronounced rise in TMA-DPH anisotropy. Our data indicate that the changes in intracellular Na + concentration, rather than those in extracellular Na + concentration, are responsible for the alterations in platelet membrane fluidity probed by TMA-DPH.

Delayed K+ current and external K+ in single cardiac Purkinje cells.

The effect of external K+ on the delayed K+ current was investigated in rabbit single Purkinje cells. Whole cell voltage clamp and intracellular dialysis were used. At K+ concentrations less than 1 mM the kinetics of the delayed K+ current were not changed, but the conductance was markedly reduced. This effect was due to a direct change at an extracellular site and not due to secondary changes in intracellular Na+ or Ca2+ concentrations. A rise in intracellular Na+ or Ca2+ rather increased the delayed K+ current. The decrease in the delayed K+ current in low external K+ was absent when the experiments were done in Na+-free solution. It is concluded that external Na+ exerts an inhibitory effect on the conductance of the delayed K+ current.

Effect of respiratory acidosis on hypoxic newborn myocardium

We studied the effect of respiratory acidosis (pH = 6.8) on mechanical function, tissue adenosine triphosphate (ATP), and effluent creatine kinase (CK) in isolated arterially perfused hypoxic newborn and adult rabbit hearts. In the oxygenated muscle, acidosis reduced tension (T) and maximal tension first derivative [ + dT dt ( max) ] in the adult more than in the newborn. In the adult hypoxic and reoxygenated hearts, acidosis during hypoxia (not reoxygenation) improved the recovery of T, + dT dt ( max) and tissue adenosine triphosphate (ATP) and reduced CK release and the rise in the resting tension. In the newborn heart, respiratory acidosis during hypoxia had no beneficial effects on recovery of mechanical function, tissue ATP and CK release. The buffering capacity and sarcolemmal HNa exchange rate are both higher in the newborn heart than in the adult heart. This suggests that acidosis reduces the rise in intracellular Na and Ca, that is observed during hypoxia and reoxygenation, in the adult more than in the newborn and this may explain the beneficial effect of acidosis in the adult and not in the newborn.

The packing of acetylcholine into quanta at the frog neuromuscular junction is inhibited by increases in intracellular sodium.

Pretreatment with hypertonic solutions, insulin, or adrenaline increases the size of quanta at the frog neuromuscular junction, as determined by measurements of miniature end plate potentials or currents (Van der Kloot and Van der Kloot 1985, 1986). The increase in quantal size apparently is due to an increase in acetylcholine (ACh) content of individual quanta. These treatments, therefore, can be used to study the packaging of ACh. Previously, I reported that increases are blocked by an inhibitor of active ACh uptake into vesicles (Van der Kloot 1986b, 1987b). The present study shows that the increases in quantal size were antagonized by inhibiting the Na+-K+ exchange pump with 100 microM ouabain, 10 microM dihydroouabain, or K+-free solutions. The increases in quantal size were also antagonized by 10 microM monensin, a Na+ ionophore, or by 5 microM aconitine, which opens Na+ channels at normal resting potentials. Apparently a rise in intracellular [Na+] inhibits the addition of ACh to quanta. The mechanism by which a rise in intracellular Na+ inhibits ACh packing is unknown, but apparently it is not due to inhibition of choline reuptake into the terminals. Also consistent with the above hypothesis is that the increase in quantal size following depolarization for 2 h in elevated [K+]out was substantially enhanced when tetrodotoxin (TTX) was present, suggesting that in the absence of TTX there is a rise in [Na+]in that antagonizes the incorporation of additional ACh into the quanta.

Regulation of intracellular pH in sheep cardiac Purkinje fibre: interactions among Na+, H+, and Ca2+1.

Experiments were performed on sheep cardiac Purkinje fibres using pH- and sodium-selective microelectrodes, while simultaneously measuring tension, to determine if the fall in intracellular pH (pHi) following a rise in intracellular Na+ activity (aiNa) is caused by inhibition or reversal of acid extrusion on Na+-H+ exchange. A rise in aiNa was induced either by using the cardioactive steroid strophanthidin to inhibit the sarcolemmal Na+-K+ pump or by increasing the frequency of stimulation (0-4 Hz). Both of these manoeuvres led to an increase in aiNa and a decrease in pHi. Following exposure to strophanthidin, amiloride (an inhibitor of sarcolemmal Na+-H+ exchange) produced a decrease in both pHi and aiNa. These effects of amiloride increased with decreasing pHi, indicating that acid extrusion on Na+-H+ exchange is stimulated by the fall in pHi. The changes in intracellular Na+ and H+ caused by amiloride were quantitatively consistent with an electroneutral stoichiometry. The fall in pHi during strophanthidin exposure is therefore not caused by inhibition or reversal of acid extrusion Na+-H+ exchange. It is likely that the fall in pHi during a rate increase is also independent of Na+-H+ exchange. This is because (i) it has been shown previously to occur in the presence of amiloride and (ii) the calcium antagonist D600 completely abolished the stimulation-dependent fall in pHi. It is concluded that the intracellular acidosis following inhibition of the sarcolemmal Na+-K+ pump or following an increase in the rate of stimulation is secondary to a rise in intracellular calcium.

PH regulation in the vertebrate central nervous system: microelectrode studies in the brain stem of the lamprey.

Studies of intracellular pH (pHi) in nervous tissue are summarized and recent investigation of intracellular and extracellular pH (pHo) in the isolated brain stem of the lamprey is reviewed. In the lamprey, pHi regulation was studied in single reticulospinal neurons using double-barrel ion-selective microelectrodes (ISMs). In nominally HCO3(-)-free HEPES-buffered media, acute acid loading was followed by a spontaneous recovery of pHi requiring 10-20 min and was associated with a prolonged rise in intracellular Na+. The recovery of pHi was blocked by 1-2 mM amiloride. Amiloride also caused a small rise in pHo. Substitution of external Na+ caused a slow intracellular acidification and extracellular alkalinization. Return of external Na+ reversed these effects. Transition from HEPES to HCO3(-)-buffered media increased the rate of acid extrusion during recovery of pHi. Recovery in HCO3(-)-buffered media was inhibited by 4,4'-diisothio-cyanostilbene-2,2'-disulfonic acid and was slowed after exposure to Cl(-)-free media. Following inhibition of acid extrusion by amiloride, transition to HCO3- media restored pHi recovery. These data indicate that lamprey neurons recover from acute acid loads by both Na+-H+ exchange and an independent HCO3(-)-dependent mechanism. Evidence for HCO3(-)-dependent acid extrusion in other vertebrate cells and the protocols of pHi studies using ISMs are discussed.

Regulation of transmembrane electrical potential gradient in rat hepatocytes in situ.

The transmembrane electrical potential gradient (Em) has been measured in hepatocytes from intact anesthetized rats using conventional intracellular microelectrodes under a variety of conditions. Em measurements in control animals were normally distributed around a mean of -35.5 +/- 4.6 mV (SD) with a coefficient of variation (CV) of 13.1% and a range of -26 to -54 mV. In individual livers, however, measurements of Em at a given point in time exhibited little cell-to-cell variation (cv of 4.5%). The Em was noted to fluctuate spontaneously over time and to change consistently in response to a variety of physiological stimuli including fasting (depolarization to -28.5 +/- 3.8 mV) and infusion of glucagon in physiological amounts (hyperpolarization to -45.0 +/- 1.8 mV). Hepatocyte Em abruptly depolarized (2-5 mV) after an intravenous bolus of taurocholate (3 mumol) or alanine (45 mumol), suggesting that both solutes exhibit electrogenic uptake. The Em returned to or below preinfusion values within 5 min. Continued infusion of alanine (10.8 mumol/min), but not taurocholate (810 nmol/min), caused a sustained and unexpected hyperpolarization of Em of 8.2 +/- 3.1 mV that lasted at least 60 min. In separate studies, alanine administration did not alter the biliary excretion of a taurocholate load. Taken together, these observations demonstrate that rat hepatocytes in situ are tightly coupled electrically and that physiological stimuli, including fasting, glucagon, and sodium-coupled solute uptake can change Em considerably over time. The late hyperpolarization of Em caused by alanine appears to offset the rise in intracellular Na+ associated with alanine uptake and preserve the Na+ electrochemical gradient such that Na+-coupled taurocholate transport is maintained.

Frequent stimulation of the guinea-pig myocardium raises the inotropic efficacy of tissue-bound ouabain.

3H-Ouabain binding to frequently (1 Hz) stimulated papillary muscles from reserpine-pretreated guinea pigs was evaluated at ouabain concentrations of 18.5 and 200 nmol/l. Myocardial activity increased the amount of 3H-ouabain bound to the tissue in comparison with quiescent preparations. Since the shape of the time course of ouabain binding changed with frequent stimulation, a greater number of ouabain-accessible binding sites of the Na pump as induced by the rise in intracellular Na with frequent stimulation cannot be the sole mechanism of the frequency dependence. In view of their stimulatory properties on the Na pump the effects of intracellular Na and extracellular K could be equivalent. By contrast, both interventions were differently effective. The K antagonism on 3H-ouabain binding was independent from stimulation frequency. Furthermore, the shape of the time course of binding was not altered by [K]o. As evidenced by the dependence of half-times to steady-state effect on muscle diameter, the apparent rate of diffusion of ouabain was accelerated with the frequency of contractions. This acceleration could have interfered with the time course of binding at frequent stimulation. After correlating the time courses of positive inotropic effect and ouabain binding (concentration of ouabain in the medium 200 nmol/l), frequent stimulation was found to raise the inotropic efficacy of tissue-bound ouabain. The relation of excitation-dependent Na influx to the saturable, ouabain-inhibited, Na pump explained the frequency dependence of the intropic efficacy of ouabain; that is, the observed change of efficacy was consistent with Na-pump saturation in dependence on intracellular Na.

Stimulation of Na,K-activated adenosine triphosphatase and active transport by low external K+ in a rat liver cell line.

Exposure of ARL 15 cells, an established line from adult rat liver, to concentrations of external K+ below 1 mM caused a rapid fall in intracellular K+ and a corresponding rise in intracellular Na+ that became maximal within 12 h. Upon continued exposure to low external K+, these initial changes were followed by a striking recovery such that, by 24 h, intracellular Na+ and K+ concentrations approached their control values. Concomitant with this recovery, there was a substantial increase in Na,K-ATPase specific activity that was detectable at 12 h and maximal at 24 h. After restoration of the external K+ concentration, the elevated level of enzyme activity showed little change for at least 24 h. In contrast, restoration of external K+ resulted in a rapid rise in intracellular K+ and a fall in Na+ such that within 30 min the Na+/K+ ratio was lower than in control cells. This overshoot, together with a demonstrated increase in active 86Rb+ uptake under "Vmax" conditions, confirms that the enhancement in Na,K-ATPase specific activity in response to low external K+ represents an increase in functional Na,K pumping capacity.

Decreased intracellular Na+ concentration is an early event in murine erythroleukemic cell differentiation.

A decrease in Na+/K+-pump activity is an early event of Friend murine erythroleukemic (MEL) cell differentiation along the erythroid pathway. This decreased Na+/K+-pump activity has been proposed to be an essential step in differentiation which would cause a rise in intracellular Na+ concentration and then, by means of Na+/Ca2+ exchange, an increase in intracellular Ca2+. An increase in intracellular Ca2+ has been proposed to be essential for induction of differentiation. A critical prediction of this Na+-Ca2+ hypothesis is the rise in intracellular Na+. To test this prediction we have measured intracellular Na+ using a novel triple isotope method involving 3H2O, [14C]sucrose, and 22Na to measure total water, extracellular fluid, and Na+, respectively. 22Na equilibration occurred in less than 10 min. In uninduced cells, intracellular Na+ was 15.2 +/- 2.2 mM (S.D., n = 22); after induction for 14-16 h with dimethyl sulfoxide, intracellular Na+ decreased significantly (p less than 0.0001) to 8.4 +/- 1.4 mM (n = 21). The time course of the decline in intracellular Na+ paralleled that of the decrease in the Na+/K+-pump activity. These results are in direct contradiction to the Na+-Ca2+ hypothesis and suggest that observed changes in Na+/K+-pump activity can be explained solely on the basis of changes in intracellular Na+. The drop in intracellular Na+ is due to a decrease in Na+ influx. We suggest, however, that the decrease in the Na+ influx is not itself an essential event of differentiation, but may be induced by a change in the flux of another ion coupled to Na+.

Stimulation of lymphocyte receptor capping by the ionophore monensin.

The carboxylic ionophore monensin has a biphasic effect on antibody-induced Thy-1 cap formation. At higher concentrations, 5 X 10(-6)-5 X 10(-5) M monesin causes a significant inhibition of receptor capping similar to that previously found with the Ca2+ selective ionophore A23187. At lower concentrations, 5 X 10(-8)-5 X 10(-7) M capping is stimulated. It is concluded that capping at lower ionophore concentrations is a specific response to the ability of monensin to induce a rise in intracellular Na+, which indirectly elevates intracellular Ca2+ activity. This in turn activates the contractile machinery required for the aggregation of surface receptors into capped structures. At higher concentrations monensin acts as a nonspecific detergent, which causes detrimental structural alterations in some of the membrane components involved in the capping process.

The intracellular sodium activity of sheep heart Purkinje fibres: effects of local anaesthetics and tetrodotoxin.

1. The intracellular Na activity (a(Na) (i)) of quiescent sheep heart Purkinje fibres has been measured using Na(+)-sensitive glass micro-electrodes. The effects of local anaesthetics (procaine and lidocaine) and tetrodotoxin (TTX) have been investigated.2. Local anaesthetics reduced the steady-state level of the intracellular Na activity in a dose-dependent manner. The highest concentrations used (10(-2)M) reduced the intracellular Na activity by about 25%.3. TTX decreased the steady-state level of the intracellular Na activity. At a concentration of 10(-6) g/ml. (3.13 x 10(-6)M), TTX produced a decrease in intracellular Na activity of approximately 10%.4. The initial rate of rise of the intracellular Na activity upon addition of the cardioactive steroid strophanthidin (10(-5)M) was used to estimate the net passive Na influx.5. Procaine (5 x 10(-4)M) caused a 50% reduction of this rate of rise of the intracellular Na activity. The highest concentration of procaine used (10(-2)M) decreased the rate of rise by approximately 80%.6. Procaine (5 x 10(-3)M) also reduced the rate of rise of intracellular Na produced by the removal of external K (K(o)), and prevented the large depolarization associated with the absence of K(o).7. TTX also produced a decrease in the rate of rise of the intracellular Na activity that occurs upon addition of strophanthidin. A maximum effect was produced in our experiments at a TTX concentration of 10(-6) g/ml. At this concentration the rate of rise of intracellular Na activity was reduced by approximately 40% at a membrane potential of -70 mV.8. We conclude from our experiments that the effects of local anaesthetics and TTX on the intracellular Na activity are brought about by a reduction of the Na permeability of the cell membrane, and that at the normal resting potential, Na entry through TTX-sensitive channels contributes greatly to the total net Na influx.

Direct measurement of changes in sodium pump current in canine cardiac Purkinje fibers

Purkinje fibers from dog hearts may have either a "high" resting potential of about -90 mV or a "low" resting potential of about -40 mV when immersed in low-Cl(-) solution containing 4 mM K(+). Brief exposure of Purkinje fibers at the low level of resting potential to K(+)-free fluid causes further depolarization, and return to K(+)-containing solution elicits a transient hyperpolarization which reaches a peak within a few seconds and then declines within a few minutes. Repeating these changes in K(+) concentration after clamping the membrane potential at its steady resting level in K(+)-containing fluid allows the changes in net membrane current presumably underlying the depolarization and transient hyperpolarization to be measured. Net inward current is recorded when the fiber is exposed to K(+)-free solution, and a transient net outward current arises when it is returned to K(+)-containing solution. The transient net outward current reflects a temporary increase in the rate of electrogenic Na(+) extrusion caused by the rise in intracellular Na(+) concentration that occurs while the sodium pump is slowed in K(+)-free fluid. Sodium extrusion remains enhanced, presumably until the internal Na(+) concentration has been brought back to its resting level. The transient outward current is completely abolished by the cardiac steroid acetylstrophanthidin, and its amplitude is increased as the prior exposure to K(+)-free fluid is prolonged. The decay of the transient outward current and the decline in intracellular Na(+) concentration both appear to follow first-order kinetics.

Sodium and Potassium Changes in Blood Bank Stored Human Erythrocytes

Storage of red cells for three weeks at 4 C under blood bank conditions resulted in a rise in intracellular Na+ and a fall in intracellular K+ with concomitant opposite changes in Na+ and K+ levels in the suspending plasma. A decline in red blood cell ATP during the storage period did not appear to be contributing to the changes. Increasing red blood cell ATP to levels 2 to 3 times normal did not prevent the cation changes from occurring. When assayed at 37 C in the presence of added Mg++, ouabain-sensitive membrane ATPase activity and kinetics of activation by Na+ were unaffected by the three week period of cold storage. However, when assayed at 4 C without added Mg++, simulating the conditions of storage, ATPase activity was negligible. Sodium and potassium did not change when red blood cells with normal ATP content were stored at 20 to 24 C even in the absence of added Mg++. Thus, a major cause for the development of cation changes in the red blood cell during blood bank storage in the temperature which inhibits membrane ATPase, allowing cations to leak unopposed into and out of the red blood cells.

Increased intracellular sodium mimics some but not all aspects of photoreceptor adaptation in the ventral eye of Limulus.

The effects of the intracellular iontophoretic injection of Na+ ions have been quantitatively compared with adaptation in ventral photoreceptors of Limulus. We find that: (a) both light adaptation and sodium injection are associated with a decrease in the variability of the threshold response amplitued; (b) both light adaptation and sodium injection are associated with a decrease in the absolute value of the temporal dispersion of the threshold response time delay; (c) the same template curve adequately fits the intensity response relationships measured under light adaptation and Na+ injection; (d) both light adaptation and Na+ injection produce a fourfold decrease in response time delay for a desensitization of 3 log units; (e) the time coures of light adaptation and dark adaptation is significantly faster than the onset of and recovery from desensitization produced by Na+ injection; (f) unlike local illumination, Na+ injection does not produce localized desensitization of the photoreceptor. These findings suggest that a rise in intracellular Na+ concentration makes at most only a minor contribution (probably less than 5%) to the total adaptation of these receptors in the intensity range we have examined (up to 3 log units above absolute threshold). However, changes in intracellular Na+ concentration may contribute to certain components of light and dark adaptation in these receptors.

The action of ouabain in promoting the release of catecholamines.

It is suggested that ouabain promotes catecholamine release by causing a rise in intracellular Na+ which, in turn, causes an elevated steady-state level of intracellular Ca2+. It is suggested that the Na+-K+-ATPase is not directly involved in exocytosis at either adrenergic or cholinergic synapses.