Efflux Rate Coefficient Research Articles

Some Effects of Nicorandil on the Smooth Muscles of the Rat and Guinea Pig

In rat isolated portal vein, nicorandil (0.1-500 microM) abolished spontaneous tension waves and inhibited mechanical responses to norepinephrine (0.1-100 microM) and KCl (5-80 mM). Intracellular electrical recording showed that nicorandil (0.1-1 microM) abolished spontaneous multispike complexes and at higher concentrations (up to 500 microM) raised the membrane potential to approximately -90 mV. Using 86Rb as a K+-marker, nicorandil (5-500 microM) increased the 86Rb efflux rate coefficient. In rat isolated aorta, nicorandil (8-32 microM) inhibited mechanical responses to norepinephrine (0.125-100 microM) and KCl (5-80 mM), but had no effect on 86Rb exchange. In guinea pig isolated taenia caeci, nicorandil (4-64 microM) relaxed spontaneous mechanical tone and increased 86Rb efflux in the absence and presence of apamin, 100 nM. It is concluded that the inhibitory effects of nicorandil in portal vein and taenia caeci are mediated at least in part by a mechanism which involves the opening of apamin-insensitive, 86Rb-permeable K+ channels. In aorta, however, the opening of such channels was not detected, and the inhibitory effects of nicorandil in this tissue are associated with an, as yet, undefined mechanism.

Taurine transport associated with cell volume regulation in flounder erythrocytes under anisosmotic conditions.

The taurine transport of flounder erythrocytes is associated with a cell volume regulation in anisosmotic media. An osmolality reduction leads to a cell volume increase, which is followed by a volume readajustment towards the original level. A 75 mosM reduction is accompanied by a 33 mumol g dry wt.-1 reduction in the cellular taurine content. The reduction in osmolality activates the taurine release mechanism by transiently increasing the rate coefficient for taurine efflux. The rate coefficient for taurine influx is similarly stimulated. This influx is mediated by a Na+-independent transport system. The concomitant activation of influx and efflux suggests a coupling between these two systems. Higher taurine efflux and influx rate coefficients which decayed more slowly with time were measured in cells suspended in Na+-free (choline replacement) media than in the presence of Na+. This suggests that Na+ may play a role in the taurine release mechanism. Noradrenaline induced a cellular swelling at normal osmolality (330 mosM), but had only a minor effect on the taurine efflux and influx and the cellular taurine content. Urea-induced cellular swelling at normal osmolality initiated a volume regulatory process and activated the taurine release mechanism, similarly to an osmolality reduction. These results show that osmolality reduction and cellular swelling are no prerequisites for the activation of the taurine release mechanism and the cell volume readajustment. It is suggested that the dimension of an intracellular solute compartment determines the activation level of this mechanism.

The effects of BRL 34915 and nicorandil on electrical and mechanical activity and on 86Rb efflux in rat blood vessels

The effects of the antihypertensive agent BRL 34915 on a variety of responses of the aorta and portal vein of the rat have been compared with those of nicorandil. On portal vein, BRL 34915 (0.01-50 X 10(-6) M) and nicorandil (0.1-500 X 10(-6) M) abolished spontaneous mechanical activity and reduced mechanical responses to noradrenaline (0.1-100 X 10(-6) M) and K+ (5-20 X 10(-3) M) but had little inhibitory effect on responses to K+ (40-80 X 10(-3) M). The onset of the reduced responses to noradrenaline was delayed by both agents. On portal vein, BRL 34915 (0.1-50 X 10(-6) M) and nicorandil (0.5-500 X 10(-6) M) abolished spontaneous electrical and mechanical activity, hyperpolarized the smooth muscle cells to a value close to their calculated potassium equilibrium potential and increased the 86Rb efflux rate coefficient. On aorta, BRL 34915 (0.2-0.8 X 10(-6) M) and nicorandil (8-32 X 10(-6) M) reduced mechanical responses to noradrenaline (0.001-1 X 10(-6) M) and K+ (5-20 X 10(-3) M) but had little inhibitory effect on responses to K+ (40-80 X 10(-3) M). On aorta, BRL 34915 (0.2-0.8 X 10(-6) M) increased the 86Rb efflux rate coefficient whereas nicorandil (8-32 X 10(-6) M) was without effect. It is concluded that the inhibitory actions of BRL 34915 on both aorta and portal vein result from the opening of membrane potassium channels. The resulting membrane shunt inhibits the effects of excitatory agents. The inhibitory effects of nicorandil result from a combination of the opening of potassium channels together with an additional, undefined action.

Effect of apamin on responses to BRL 34915, nicorandil and other relaxants in the guinea‐pig taenia caeci

BRL 34915 (4-64 X 10(-7) M), isoprenaline (0.5-32 X 10(-8) M) and nicorandil (4-64 X 10(-6) M) produced a slowly-developing relaxation of spontaneous tone of the guinea-pig taenia caeci; with no after-contraction on washout. These inhibitory responses were unaffected by apamin (10(-7) M). Adenosine triphosphate (0.06-2 X 10(-3) M) and noradrenaline (1-16 X 10(-7) M) produced a rapid inhibition of spontaneous tone with a prominent after-contraction, especially on washout. Both the inhibitory effect and the rebound contraction were abolished by apamin (10(-7) M). Exposure to both BRL 34915 (64 X 10(-7) M) and to nicorandil (64 X 10(-6) M) produced an increase in the 86Rb efflux rate coefficient which was unaffected by apamin (10(-7) M). Exposure to isoprenaline (32 X 10(-8) M) had no effect on the 86Rb efflux rate coefficient. Exposure to noradrenaline (16 X 10(-7) M) produced an increase in the 86Rb efflux rate coefficient which was abolished by apamin (10(-7) M). The results confirm that both BRL 34915 and nicorandil are capable of opening potassium channels in smooth muscle but show that the channel is not apamin-sensitive.

Mineralocorticoid effect on K+ permeability of the rabbit cortical collecting tubule

Mineralocorticoid hormones stimulate Na+ absorption and K+ secretion by the cortical collecting tubule. There is good evidence that this stimulation involves increasing luminal membrane Na+ permeability and the turnover rate (or number) of the Na+-K+ pumps. These experiments were designed to examine whether mineralocorticoid hormones also increase cell K+ permeability. Using 42K tracer measurements in tubules treated with amiloride to inhibit active Na+ and K+ transport, passive K+ permeation increased with increasing mineralocorticoid effect. Net Na+ absorption and the (passive) K+ efflux rate coefficient (KK) showed a linear relationship. The stimulatory effect was evident in vitro since 0.2 microM aldosterone added to the bath of tubules harvested from NaCl-loaded rabbits increased KK at 3 hrs while time controls showed no change. Since these tubules were also treated with amiloride, this increase in KK was not dependent on increasing Na+ absorption. The results indicate that in addition to the well-described effects of aldosterone on Na+ permeability and cell metabolism, the mineralcorticoid effect includes an increase in cellular K+ permeability.

Cellular K+ permeation across the cortical collecting tubule: effects of Na+-K+ pump inhibition and membrane depolarization

These experiments examined the possibility that alterations in cell cation content and/or membrane voltage could influence cell K+ permeability of the cortical collecting tubule. Using the amiloride-treated isolated perfused rabbit cortical collecting tubule, ouabain or a K+-free bath reduced the magnitude of the K+ diffusion voltage. In addition, both methods of Na+-K+-ATPase inhibition reduced the K+ efflux (lumen-to-bath) rate coefficient (KK) without affecting the Na+ efflux rate coefficient. The magnitude of the reduction of KK could not be explained by a model of simple diffusion across two membranes in series even if the intracellular voltage were abolished. Thus, pump inhibition reduced cell K+ permeability. To determine whether membrane depolarization could induce a change in membrane permeability, [K+] was increased to 20 mM in both perfusate and bath. The reduction in KK was within the range predicted by the three-compartment model (36%). Differential membrane depolarization by raising lumen [K+] or bath [K+] produced disparate results. Apical depolarization reduced KK but basolateral depolarization did not. Taken together these results indicate that intracellular ion content may play a major role in regulating cell K+ permeability independent of voltage-dependent effects. In addition, under these experimental conditions, the apical membrane may be the rate-limiting barrier to cellular transfer.

Release of gamma-aminobutyric acid from cat colon

The release of gamma-aminobutyric acid was confirmed in isolated cat colon loaded with tritiated gamma-aminobutyric acid. Thirty to 180 minutes after loading the spontaneous efflux of tritium appeared to fit a single exponential curve with an efflux rate coefficient of 0.002 per minute. Electrical stimulation produced frequency-dependent increases in the tritium efflux and in the contractions. Even 120 minutes later over 91 percent of the total radioactivity in the superfusates was attributable to tritiated gamma-aminobutyric acid. The acid release and the contractions induced by electrical transmural stimulation were inhibited by tetrodotoxin and by a calcium-free medium. Release of the acid was not significant during contractions elicited by nicotine and acetylcholine. These findings indicate that gamma-aminobutyric acid is released from the terminals of neurons in the myenteric plexus of the colon.

Effect of Na+ and ATP on peritubular Ca transport by the marine teleost renal tubule.

Ca uptake rates and efflux rate coefficients were determined in isolated renal tubules of the winter flounder, Pseudopleuronectes americanus. Na-free medium and 10(-4) M ouabain depressed while 0.2 mM dinitrophenol stimulated Ca efflux. The effects on Ca movement appeared to be associated with the adenosine triphosphate (ATP) concentration of the tubular fluid space (TFS), rather than the bath-to-tissue Na gradient. Elevation of the ATP concentration of TFS by incubation of tissue in external ATP more than doubled Ca uptake rate. Inhibition of Ca uptake by lanthanum and stimulation by Ca ionophore A23187 suggested that ATP altered plasma membrane Ca transport. Incubation in external phosphate had no effect. Plasma membrane vesicles (PMV) prepared from flounder tubules showed saturable Ca uptake inhibited by Mg, unaffected by Na gradients, and stimulated by intravesicular but not extravesicular ATP. ATP appeared to stimulate PMV Ca uptake by increasing membrane Ca binding. We concluded that Ca uptake across the peritubular membranes was not directly linked to the serosal Na gradient but was greatly influenced by the intracellular ATP concentration.

Changes in mouse oocyte membrane potential and permeability during meiotic maturation.

Mouse oocytes spontaneously undergo meiotic maturation when removed from the ovary and cultured in vitro. This paper describes the changes in cell surface membrane potential from the germinal vesicle (GV) stage (- 46 mV, inside negative), through germinal vesicle breakdown (GVBD) (- 31 mV), to the polar body (PB) stage (- 17 mV). Radioisotope tracer flux experiments established the total exchangeable internal K at the GV stage in 190 mM, the K efflux rate coefficient as 6.7 X 10(-5) sec-1) and the K permeability as 46.7 X 10(-8) cm sec-1. In comparison with previous results with PB stage oocytes, this suggests that the membrane potential depolarization is caused largely by a decrease in K permeability. When GVBD is blocked with 0.11 mM dibutyryl cyclic AMP, the membrane potential does not depolarize. This suggests a linkage between depolarization and GVBD. However, incubation in 80 mM K medium, which depolarizes the oocyte, does not trigger GVBD. It is concluded that membrane depolarization by itself is not sufficient to cause GVBD.

Na and K transport across the cortical and outer medullary collecting tubule of the rabbit: evidence for diffusion across the outer medullary portion

The rabbit collecting tubule displays functional axial heterogeneity with respect to Na ion transport. The present experiments compared cortical collecting tubule (CCT) and outer medullary collecting tubule (OMCT) Na and K transport. Na efflux across the CCT was inhibited by ouabain, whereas Na efflux across the OMCT was smaller and unaffected by ouabain. Assessment of the equivalent conductivities of Na and K across the CCT by imposition of a Na-K bi-ionic gradient demonstrated a higher K/Na conductivity across the CCT than would be predicted from their respective limiting equivalent conductivities in water. In contrast, the ratio of their conductivities across OMCT were not different than would be predicted by their ratio in water. The "selective" nature of the Na and K pathways across CCT was confirmed by measuring the tracer efflux rate coefficients. In the amiloride-treated CCT the K/Na rate coefficient ratio was 9.8 +/- 1.5; this ratio across the OMCT was 1.51 +/- 0.10. The latter value is not different from the ratio of the mobilities of these ions in water. The diffusional nature of Na and K transfer across OMCT was confirmed by the demonstration of the concentration-independent Na efflux rate coefficient and the demonstration of appropriate net Na and K transepithelial flows in response to imposition of oppositely directed chemical gradients. Although the permeability of the OMCT is low, the chemical gradients found in vivo might be sufficient to effect some K absorption and Na secretion without completely dissipating the steep gradients generated by the CCT. These transport characteristics might be important in the regulation of Na excretion and K recycling into the renal medulla.

The effect of extracellular calcium deprivation on amylase secretion and 45Ca efflux from rat pancreas.

1. The role of extracellular Ca(2+) in pancreatic enzyme secretion and (45)Ca efflux evoked by acetylcholine (ACh) and caerulein has been assessed in the incubated, uncinate pancreas of young rats.2. In a medium containing 2.5 x 10(-3)m-Ca(2+), the maximal rates of amylase secretion evoked by optimal doses of each secretagogue were similar. However, the time courses of amylase release during prolonged stimulation of the gland were different.3. The time course of amylase secretion in response to an optimal dose of ACh (10(-5)m) was characterized by an initial rapid increase followed by a slow sustained rise. For caerulein (10(-8)m), an initial rapid rise was followed either by a plateau or slight decline in the rate of amylase secretion.4. Both secretagogues produced similar increases in the rate coefficient of (45)Ca efflux from the gland.5. With supra-optimal doses of the secretagogues, amylase secretion, but not the rate coefficient of (45)Ca efflux, was depressed.6. Reducing the extracellular Ca(2+) concentration did not have a marked effect on basal amylase secretion but inhibited the action of both secretagogues. When the Ca(2+) concentration was 10(-6)m or lower, these inhibitory effects were irreversible. Amylase secretion stimulated by ACh was more sensitive to extracellular Ca(2+) deprivation than that stimulated by caerulein, the concentration required for half-maximal secretion being about 9-fold greater for ACh.7. Decreasing the extracellular Ca(2+) concentration increased both the basal and stimulated rate coefficients of (45)Ca efflux.8. Our results support the hypothesis that pancreatic enzyme secretagogues act by releasing bound Ca(2+) from sites within the acinar cell. Furthermore, they suggest that the site utilized by ACh is more amenable to depletion, via changes in extracellular Ca(2+) concentration, than that employed by caerulein.

Uptake and release of 45Ca by Myxicola axoplasm.

The binding and release of 45Ca by axoplasm isolated from Myxicola giant axons were examined. Two distinct components of binding were observed, one requiring ATP and one not requiring ATP. The ATP-dependent binding was largely prevented by the addition of mitochondrial inhibitors, whereas the ATP-independent component was unaffected by these inhibitors. The ATP-independent binding accounted for roughly two-thirds of the total 45Ca uptake in solutions containing an ionized [Ca2+] = 0.54 microM and was the major focus of this investigation. This fraction of bound 45Ca was released from the axoplasm at a rate that increased with increasing concentrations of Ca2+ in the incubation fluid. The ions Cd2+ and Mn2+ were also able to increase 45Ca efflux from the sample, but Co2+, Ni2+, Mg2+, and Ba2+ had no effect. The concentration-response curves relating the 45Ca efflux rate coefficients to the concentration of Ca2+, Cd2+, and Mn2+ in the bathing solution were S-shaped. The maximum rate of efflux elicited by one of these divalent ions could not be exceeded by adding a saturating concentration of a second ion. Increasing EGTA concentration in the bath medium from 100 to 200 microM did not increase 45Ca efflux; yet increasing the concentration of the EGTA buffer in the uptake medium from 100 to 200 microM and keeping ionized Ca2+ constant caused more 45Ca to be bound by the axoplasm. These results suggest the existence of high-affinity, ATP-independent binding sites for 45Ca in Myxicola axoplasm that compete favorably with 100 microM EGTA. The 45Ca efflux results are interpreted in terms of endogenous sites that interact with Ca2+, Cd2+, or Mn2+.

Heterogeneity of the rabbit collecting tubule: Localization of mineralocorticoid hormone action to the cortical portion

This study was designed to examine the sodium, potassium, and chloride transport rates across the cortical and outer medullary collecting tubule and to localize the action of mineralocorticoid hormone. Rabbit collecting tubules were dissected from the cortex (CCT), the outer stripe of the outer medulla (OMCT0), or the inner stripe of the outer medulla (OMCTi). From normal rabbits, the transepithelial voltage was -20.8 +/- 2.7 mV in CCT, -4.2 +/- 1.4 mV in OMCT0, and +10.6 +/- 2.3 mV in OMCTi. From DOCA-treated rabbits, only the CCT voltage was different (-42.9 mV). Net sodium absorption across the CCT increased ith DOCA-treatment from 23 +/- 5 to 54 +/- 8 pEq . mm-1 . min-1, whereas net potassium secretion increased from 15 +/- 2 to 43 +/- 5 pEq . min-1 . min-1. Net chloride absorption was significant only in DOCA-treated rabbits. The OMCTi, in contrast, displayed no net transport of sodium, potassium, or chloride and had a lower rate coefficient for sodium efflux than did the CCT. DOCA treatment had no effect on any transport rate measured in this segment. The OMCT0 has small potassium secretion, which did not increase with DOCA treatment. The collecting tubule is heterogeneous along its length with respect to ion transport. Mineralocorticoid-hormone-sensitive sodium absorption is present predominantly, if not exclusively, in the cortical collecting tubule.

Apparent affinity changes induced by insulin of Na-K transport system in frog skeletal muscle.

The dependency of Na efflux on intracellular Na concentration was examined on sartorius muscles of Rana catesbeiana. In normal Ringer, the rate coefficient of 22Na efflux is nearly proportional to the internal Na concentration, i.e., Hill's coefficient in the dependency of Na efflux on internal Na concentration was around two. As long as insulin is present in the bathing solution, the rate coefficient of 22Na efflux retains the stimulated level. Insulin caused a leftward shift of the relationship between Na efflux and the logarithm of internal Na concentration, and concomitantly decreased Hill's coefficient. A model of the coupling of Na transport and hydrolysis of ATP was proposed. The theoretical relation derived from this model accounted for these findings quite satisfactorily. According to this model, insulin shifts as a cofactor the equilibrium between dephosphorylated and phosphorylated states of carriers, whose Na binding sites are not occupied by Na, toward the phosphorylated state.

Effects of ouabain on Na efflux in high internal Na and insulin-preincubated muscles.

To gain an insight into the mechanism of stimulation by insulin of Na efflux, the effect of high concentrations of internal Na on the inhibition by ouabain of Na efflux was compared to the inhibition by ouabain of insulin-stimulated Na efflux. The rate coefficient of 22Na efflux from "high-Na" muscle exposed to ouabain was lower than that from "low-Na" muscle exposed to ouabain. Similarly, the rate of net Na loss from the "high-Na" muscle which had been exposed to ouabain was lower than that frm the "low-Na" muscle. These findings indicate that Na transport units whose internal Na-binding sites have been occupied by Na is susceptible to ouabain. The insulin-stimulated Na efflux was inhibited by ouabain to a larger extent than was the Na efflux from the control muscle, although the final level of the rate coefficient of 22Na efflux from the muscle exposed to both insulin and ouabain was not always lower than that from the muscle exposed only to ouabain. The similarity between the effects of ouabain in "high-Na" muscle and in insulin-preincubated muscle suggests that insulin increases the fraction of Na transport units whose internal Na-binding sites have been occupied by Na.

Gamma-Aminobutyric acid efflux from sympathetic glial cells: effect of 'depolarizing' agents.

1. Isolated desheathed rat superior cervical ganglia were incubated in [3H]2,3,-gamma-aminobutyric acid ([3H]GABA) solution (1--10 microM for 2--3 hr) in the presence of 10 microM-amino-oxyacetic acid (AOAA). The subsequent efflux of tritium into a stream of superfused non-radioactive GABA-free Krebs solution at 25 degrees C was measured. 2. In the presence of 10 micrometer-AOAA the mean basal efflux rate coefficient (k0) for exit of tritium into the superfusion fluid was 0.7 x 10(-3) min-1. More than 98% of effluent tritium comprised unchanged [3H]GABA. The rate coefficient showed no correlation with the amount of [3H]GABA previously accumulated by the ganglion. 3. Elevation of [K+]o to greater than 50 mM increased the rate coefficient for [3H]GABA release by up to four times. Changes in efflux rate were not correlated with osmotic changes, and persisted after re-accumulation of effluent [3H]GABA by the inward carrier was inhibited. The effect of alkali metal cations diminished in the order Rb+ greater than K+ greater than Cs+Li+. Effects of K+ solutions were not reduced by omitting Ca2+ ions, with or without the addition of Mg2+. 4. Application of electrical pulses (0.1--1 msec duration, 1--10 Hz, 4 min trains) to the ganglion soma or to the preganglionic nerve trunk also raised k0. This effect declined with repeated stimulus trains, without an accompanying diminution in the response to K+. Responses to electrical stimulation were not reduced by amethocaine (300 microM), tetrodotoxin (3 microM) or raised [Mg2+i1 (0 mM-[Ca2+]/30 mM-[Mg3+]). Separate local superfusion of the pre- and post-ganglionic nerve trunks and of the ganglion soma showed that the response to electrical stimulation was localized to the vicinity of the stimulus and was not propagated along the nerve trunks or across the synapses. 5. Electrical recording from impaled 'inexcitable' cells (presumed to be neuroglial cells (Appendix)) indicated that the quantities of K+ ion accumulating during repetitive nerve stimulation are insufficient to stimulate the release of GABA from the glial cells. No physiological role for the release process in modulating neuronal excitability could be adduced.

Saturation behavior of ascites tumor cell chloride exchange in the presence of gluconate

Steady state Cl − flux across the Ehrlich mouse ascites cell membrane was studied when gluconate replaced Cl − in the external medium. Saturation behavior was observed; K 1 2 was 23.9 mM Cl − and V was 758 μmol · g −1 dry weight · h −1. The cells lost K +, Cl − and H 2O, consistent with relative impermeability to gluconate, and the Cl − efflux rate coefficient was elevated. The results indicate that a major portion of Cl − exchange occurs as a membrane transport process and suggest that the process is sensitive to intracellular Cl − levels.

Serum‐stimulated changes in calcium transport and distribution in mouse 3T3 cells and their modification by dibutyryl cyclic AMP

Serum stimulation of quiescent 3T3 cells returns the cells to a proliferative state. Changes in Ca content, transport and distribution during the transition through G1 and S phase have been investigated following serum stimulation of these cells. 45 Ca exchange data indicate at least two kinetically defined cellular compartments for Ca; a rapidly exchanging component presumably representing surface Ca which is removable by EGTA and a slowly exchanging component presumably representing cytoplasmically located Ca. Previous studies (Tupper and Zorgniotti, '77) indicate that the approach to quiescence in the 3T3 cells is characterized by a large increase in the surface Ca component. The present data demonstrate that this component is rapidly lost following serum stimulation. Furthermore, the serum induces an 8-fold increase in Ca influx into the cytoplasmic compartment and a reduction in the unidirectional efflux rate coefficient for Ca. The increased Ca uptake peaks at approximately six hours (mid G1) and is accompanied by a parallel increase in cellular Ca. Prior to entrance of the cells into S phase (10-12 hours), Ca uptake declines. This is followed by a slower decline in cytoplasmic Ca levels. Simultaneous addition to fresh serum plus 0.5 mM dibutryl cAMP inhibits the entrance of the cells into S phase. Under these conditions the loss of surface Ca is not blocked. However, the presence of 0.5 mM dibutyryl cAMP inhibits the increase in Ca uptake and, in turn, diminishes the increase in cellular Ca following serum stimulation. In contrast, a low level of dibutyryl cAMP (0.1 mM) enhances progression through G1 phase but also reduces both Ca uptake and Ca content of the cells. The data suggest that the serum induced changes in Ca content and transport are linked to intracellular cyclic nucleotide levels and progression through G1 phase and that extracellular cAMP elevating agents may enhance of inhibit these interactions in a concentration dependent manner.

Calcium movements during the release of catecholamines from the adrenal medulla: effects of methoxyverapamil and external cations.

1. Cortex-free adrenal glands previously labelled with the isotope (45)Ca have been perfused with Locke or modified Locke solution to assess Ca(2+) movements under different conditions.2. Substitution of Na(+) by either sucrose or choline during perfusion with Ca(2+)-free Locke solution induced a significant and sustained decrease in the (45)Ca efflux. Concomitant with this effect there was an increase in the output of catecholamines from the perfused gland.3. In the presence of Ca(2+) (2.2 mM) in the perfusion fluid, Na(+) omission induced an increase in the (45)Ca efflux. This increase was significantly reduced if 3 x 10(-4)M methoxyverapamil (D-600) was present in the perfusion fluid. However, the increased catecholamine output in response to Na(+) deprivation remained unchanged.4. Excess of Mg(2+) (20 mM) in the extracellular medium blocked the increase in catecholamine output in response to Na(+) omission. However, the decrease in the (45)Ca efflux produced by Na(+) deprivation in the presence of this high concentration of Mg(2+) was similar to that observed in the presence of 1.2 mM-Mg(2+).5. In the absence of Mg(2+) in the extracellular medium, substitution of Na(+) by either sucrose or choline induced a sharp and transient increase in the (45)Ca efflux rate coefficient. This increased (45)Ca efflux, which has similar time course as the enhanced catecholamine output, was not affected by the presence of 3 x 10(-4)M methoxyverapamil.6. In the absence of Mg(2+), the graded substitution of Na(+) in the perfusion medium by sucrose enhanced the efflux of (45)Ca. This increase in the (45)Ca outward movement was linearly related to the logarithm of the extracellular Na(+) concentration.7. After perfusion of glands with Ca(2+)-free Locke solution, the reintroduction of Ca(2+) (2.2 mM) into the perfusion fluid produced an increase in the (45)Ca efflux. This was accompanied by a discharge of catecholamines.8. Although Mg(2+) (20 mM) was effective in blocking catecholamine release, this divalent cation did not modify the increase in the (45)Ca efflux produced by Ca(2+) reintroduction.9. In contrast to these later observations, methoxyverapamil (3 x 10(-4)M) was effective in inhibiting both increases in catecholamine output and (45)Ca efflux in response to Ca(2+) reintroduction.10. It is concluded from these experiments that (a) Ca(2+) movements in the adrenal medulla may involve both Na(+)-Ca(2+) and Ca(2+)-Ca(2+) exchange mechanisms; (b) the omission of Na(+) from the extracellular environment produces not only an increase in the output of catecholamines but it may increase the intracellular levels of Ca(2+) and that this may result in an increased Ca(2+) efflux when Mg(2+) is omitted from the perfusion fluids, and that (c) the competition between Ca(2+) and Mg(2+) during the secretory process may involve an intracellular site.

Reduction of potassium permeability by chloride substitution in cardiac cells.

1. The efflux of radioactive K and Cl ions was measured in cow Purkinje fibres and ventricular preparations of cow, cat and frog. The effect of K and Cl was studied by changing the extracellular K concentration between zero and 54 mM, and by substituting Cl ions by acetylglycinate, isethionate, benzenesulphonate, propionate and nitrate. 2. In the absence of Cl the rate coefficient for 42K efflux showed a pronounced fall, which was more pronounced the higher the K concentration. This effect was not related to the change in membrane potential. The rate coefficient for 42K efflux increased in the presence of higher extracellular K concentrations. 3. 36Cl efflux increased in the presence of reduced as well as increased extracellular K concentrations. 4. The calculated permeability coefficient for K (PK) was maximal at 5-4 mM-K, decreased slightly at higher K concentrations, but fell markedly in K-free solutions, (to about 1/5 of the value in 5-4 mM-K). In Cl-free medium PK reduced to 0-67 of the value in the Cl medium, irrespective of the K concentration. 5. The calculated PCl was greater in K-free and 16-2 mM-K than in 5-4 mM-K. 6. The ratio PK/PCl showed important changes as a function of extracellular K concentration: the value was 5 in 5-4 mM-K and fell to 2 and 0-5 respectively in 16-2 and K-free solutions. 7. The results suggest that part of the changes in membrane resistance measured by electrical methods in Cl-free media is due to a simultaneous decrease in K conductance.