A description of the nicotine stimulus and tests of its generalization to amphetamine

The mechanism responsible for the inter-individual differences of the Li⁺ distribution between red cells and plasma was studied in vitro on erythrocytes of Li⁺-treated patients showing marked differences in the in vivo distribution ratio Li _i ⁺ /Li _e ⁺ .Li⁺ distribution ratios determined after 24 h of incubation under standardized conditons were highly correlated with the steady-state in vivo Li⁺ ratios.Rates of Li⁺ transfer mediated by the Na⁺-dependent Li⁺ countertransport system (Na⁺-dependent Li⁺ release, Li⁺ uphill transport into the medium, ouabain-insensitive Li⁺ uptake) were reduced in erythrocytes exhibiting high in vivo Li⁺ ratios as compared to cells characterized by low Li⁺ distribution ratios.Variations in ouabain-sensitive Li⁺ uptake, red cell Na⁺ content, and in the ratio Na _i ⁺ /Na _e ⁺ appear not to be essentially involved in the establishment of markedly different in vivo Li⁺ ratios.It is concluded that variations in the efficiency of the Na⁺-dependent Li⁺ countertransport system are responsible for the considerable interindividual differences in the in vivo Li⁺ distribution ratio observed among Li⁺-treated patients.     

Effects of lithium and thallous ions on sodium pump activity in the guinea-pig heart and their relationship to the positive inotropic action

Summary It has recently been demonstrated that both Tl⁺ and Li⁺ produce concentration-and time-dependent positive inotropic effects in guinea-pig atrial preparations although Tl⁺ inhibits and Li⁺ stimulates isolated Na⁺, K⁺-ATPase in vitro. In order to elucidate the mechanism of the positive inotropic actions of these cations, the effects of Tl⁺ and Li⁺ on sodium pump activity were studied. Active ⁸⁶Rb uptake in guinea-pig ventricular slices, an estimate of sodium pump activity, was highly sensitive to the inhibitory effect of the cardiac glycosides. Preincubation of slices with Tl⁺ caused a dose-and time-dependent inhibition of active ⁸⁶Rb uptake. Similar concentration-and time-dependent inhibition of active ⁸⁶Rb uptake was observed when Na⁺ in a Krebs-Henseleit solution was partially replaced with Li⁺. Lithium, however, stimulated a partially purified Na⁺, K⁺-ATPase in vitro. During heart slice incubation, Tl⁺ and Li⁺ accumulated in a time-dependent manner. This accumulation was not readily reversible when slices were transferred into Tl⁺-or Li⁺-free solutions. It appears that the inhibition of sodium pump activity is related to the positive inotropic action of these cations.This work was supported by United States Public Health Service Grants HL-16052 and HL-16055 and by the Michigan Heart Association. A part of this study was presented at the 60th Annual Meeting of the Federation of American Societies for Experimental Biology, Anaheim, California, April 1976     

Cholinergic regulation of Na+-K+-ATPase activity in rat parotid gland: changes after castration

In this study, we investigated the different signalling pathways involved in muscarinic acetylcholine M(3) receptor-dependent modulation of Na(+)-K(+)-ATPase in parotid glands from normal and castrated rats. Carbachol inhibited the enzyme activity in parotid glands from control rats while it stimulated the enzyme activity in castrated rats. The inhibition of Ca(2+) calmodulin by trifluoperazine abolished the inhibitory effect of carbachol in control rats, while the inhibition of protein kinase C by staurosporine stimulated Na(+)-K(+)-ATPase. In castrated rats, trifluoperazine inhibited the carbachol-stimulant effect while staurosporine had no effect. Results indicate that in control glands the activation of a phospholipid-Ca(2+) calmodulin-dependent protein kinase C is responsible for the inhibitory effect of carbachol on Na(+)-K(+)-ATPase activity. In castrated rats, the activation of the enzyme by carbachol is regulated by its Ca(2+) calmodulin-stimulating action, and not by activation of protein kinase C. The activation of the Na(+)-K(+)-ATPase observed in castrated rats resulted in a decrease in carbachol-induced net K(+) efflux and thereby could decrease salivary fluid production.     

Angiotensin II-induced increase in slowly exchanging 45Ca2+ in relation to contractile responses of rat and guinea-pig aorta

Summary To gain more information about sources of activator Ca²⁺ involved in the contraction of rat and guinea-pig aorta evoked by angiotensin II and their sensitivity to Ca ²⁺ entry blockers, measurement of slowly exchanging ⁴⁵Ca²⁺ was established. A more physiological procedure was used, replacing La³⁺- and EGTA- containing solutions by a normal Ca²⁺-containing buffer. It was demonstrated that the angiotensin 11-induced increase in slowly exchanging ⁴⁵Ca²⁺ in rat aorta was incompletely (by approximately 60%–70%) inhibited by the organic Ca²⁺ entry blockers nifedipine, verapamil and diltiazem and by other Ca⁺ entry blocking compounds like CoCl₂ and chlorpromazine. 8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride (TMB-8) was able to inhibit the angiotensin II-induced increase in ⁴⁵Ca²⁺ content completely, but this may be an intracellular storage effects. By contrast, the organic Ca²⁺ entry blockers completely inhibited that part of the angiotensin II-induced contraction of rat aorta which was dependent upon extracellular Ca²⁺.In guinea-pig aorta, the increase in ⁴⁵Ca²⁺ content elicited by angiotensin 11 could be completely suppressed by all compounds under study. The results of these experiments correlated well with data from the functional experiments in guinea-pig aorta. In both preparations the release of Ca ²⁺ from a rapidly as well as a slowly exchanging intracellular pool appears to contribute to the contractile response elicited by angiotensin 11. Offprint requests to P. N. M. van Heiningen at the above address     

Inhibition of Na+ channel currents in rat myoblasts by 4-aminopyridine

Our previous study revealed that 4-aminopyridine (4-AP), a specific blocker of A-type current, could also inhibit inward Na+ currents (I(Na)) with a state-independent mechanism in rat cerebellar granule cells. In the present study, we report an inhibitory effect of 4-AP on voltage-gated and tetrodotoxin (TTX)-sensitive I(Na) recorded from cultured rat myoblasts. 4-AP inhibited I(Na) amplitude in a dose-dependent manner between the concentrations of 0.5 and 10 mM without significant alteration in the activation or inactivation kinetics of the channel. By comparison to the 4-AP-induced inhibitory effect on cerebellum neurons, the inhibitory effect on myoblasts was enhanced through repetitive pulse and inflected by changing frequency. Specifically, the lower the frequency of pulse, the higher the inhibition observed, suggesting that block manner is inversely use-dependent. Moreover, experiments adding 4-AP to the intracellular solution indicate that the inhibitory effects are localized inside the cell. Additionally, 4-AP significantly modifies the properties of steady-state activation and inactivation kinetics of the channel. Our data suggest that the K+ channel blocker 4-AP inhibits both neuron and myoblast Na+ channels via different mechanisms. These findings may also provide information regarding 4-AP-induced pharmacological and toxicological effects in clinical use and experimental research.     

The Na+/H+ exchanger modulates long-term potentiation in rat hippocampal slices

Although present in great variety in the brain, the role of Na⁺/H⁺ exchangers (NHEs) in hippocampal plasticity is still unknown and the effect of NHE inhibition on long-term potentiation (LTP) has not been studied yet. As it is conceivable that NHE inhibitors may severely affect mechanisms that are considered to underlie learning and memory we investigated whether the broad-spectrum NHE inhibitor 5′-(N-ethyl-N-isopropyl)-amiloride (EIPA, 10 μM) influences LTP induced by different stimuli based on a theta burst in interface hippocampus slices from 7–8-week-old Wistar and 30-month-old Fischer 344/Brown–Norway F1 hybrid (F344/BN) rats. EIPA did not affect basal synaptic transmission, paired pulse inhibition, or LTP induced by a weak stimulus, but improved the maintenance of the LTP of the population spike induced by a strong tetanus. Our data suggest that NHE activity serves as a negative feedback mechanism to control neuronal excitability and plasticity in both young and senescent animals.     

Characteristics and possible mechanisms of low - Na+ induced contractions in rat aorta

The influence of reducing external Na⁺ concentration ([Na⁺]^ex) upon vascular smooth muscle contractility was investigated using the rat isolated aorta. NaCl from the physiological saline solution (PSS) was replaced with either choline-Cl, sucrose, or LiCl to give the following [Na⁺]_ex (mM): 115, 85, 55, and 25 (115NaPSS to 25NaPSS). Small reductions in [Na⁺]_ex (115NaPSS) induced a biphasic contraction, comparable in amplitude with the control one induced by phenylephrine 10^–6 M. Elimination of the endogenous catecholamine participation using either phentolamine 10^–5 M or guanethidine 3.10^–6 M similarly reduces these contractions to 25% (sucrose replacement). A similar relaxing effect was obtained with D600 10^–5 M, an antagonist of the voltage operated Ca²⁺ channels (25–30% residual tension for all the substitutes). Large reductions in [Na⁺]_ex (25NaPSS) induced contractions comparable in amplitude and shape, but less sensitive to phentolamine and guanethidine (residual tension 65–75 %, sucrose replacement) and insensitive to D600 (all the substitutes). The Na⁺/K⁺ ATPase inhibitor ouabain (10^–4 M) elicited slowly developing contractions, the amplitude being 115% of the phenylephrine 10^–6 M control.Phenylephrine further contracted the 115NaPSS precontracted preparations, but was significantly less effective in 25NaPSS, although the precontraction levels were similar for the same substitute used. The amplitude of the superimposed phenylephrine contractions exhibited [Na⁺]_ex dependence. Phenylephrine 10^–6 M failed to further contract the ouabain 10^–4 M precontracted rings.We conclude that relatively small reductions in [Na⁺]_ex are able to induce contractions of rat aorta primarily through release of endogenous catecholamines, probably through neural Na⁺/Ca²⁺ exchange. Larger reductions in [Na⁺]_ex appear to cause contraction through muscular Na⁺/Ca²⁺ exchange.     

Role of Na+ and protein kinase C in angiotensin desensitization and tachyphylaxis in the guinea-pig ileum

Summary Simultaneous recordings of the tension and intracellular Ca²⁺ concentration of guinea-pig ileum longitudinal smooth muscle strips, as well as ²⁴Na⁺ and ⁴⁵Ca²⁺ influx measurements in cultured myocytes from the same tissue, were used to investigate the mechanisms underlying angiotensin-induced desensitization and tachyphylaxis. Angiotensin II and [2-lysine]-angiotensin II (Lys²All), incubated for prolonged periods (10 min) with muscle strips, induced fading of the contractile response (desensitization) and reappearance of the intracellular Ca²⁺ concentration oscillations, which were inhibited during the initial increase in cytosolic Ca²⁺. The desensitization was paralleled, in cultured myocytes, by inhibition of the ⁴⁵Ca²⁺ but not of the ²⁴Na⁺ influxes which were initially stimulated by the peptides. On the other hand, repeated administrations of angiotensin II (but not of Lys²All) caused gradual reduction of the contractile response and of the ²⁴Na⁺ influx stimulation evoked by the agonist (tachyphylaxis). Treatment with phorbol 12–13 dibutyrate accelerated the desensitization induced by both angiotensin II and by Lys²All and aggravated the tachyphylaxis to angiotensin II. The results support the hypothesis that activation of protein kinase C is responsible for the desensitization and that tachyphylaxis is due to the slow dissociation of angiotensin II from a postulated Na⁺-dependent regulatory site on the receptor.Correspondence to S.I. Shimuta at the above address     

中药大黄的生化学研究ⅩⅫ蒽醌衍生物对兔肾髓质Na+-K+-ATP酶活性的抑制和利尿作用

家兔实验表明:大黄素、大黄酸以30 mg/kg的剂量灌胃给药,2～4h后尿量、排Na⁺和K⁺量达最高峰,比对照组明显增多。而芦荟大黄素和大黄酚的作用较弱。大黄素、大黄酸和芦荟大黄素对免肾髓质Na⁺-K⁺-ATP酶活性有较强的竞争性抑制作用。     

Effects of monovalent cations on cardiac Na+, K+-ATPase activity and on contractile force

Summary The relationship between Na⁺, K⁺-ATPase inhibition by monovalent cations and their inotropic effect was studied in guinea pig hearts. The activity of partially purified cardiac enzyme was assayed in the presence of 5.8 mM KCl and either 20 or 150 mM NaCl. Rb⁺ and Tl⁺ inhibited Na⁺, K⁺-ATPase activity, the magnitude of the inhibition by these cations being greater in the assay media containing lower Na⁺ concentrations. Tl⁺ produced a dose-dependent inhibition of Na⁺, K⁺-ATPase activity in the presence of 20 mM Na⁺ and 75 mM K⁺, a cationic condition similar to that of intracellular fluid. Other monovalent cations such as K⁺, Cs⁺, NH₄ ⁺, Na⁺ or Li⁺ produced essentially no effect on the Na⁺, K⁺-ATPase activity or slightly stimulated it. In left atrial strips stimulated with field electrodes and bathed in Krebs-Henseleit solution (5.8 mM K⁺ and 145 mM Na⁺), addition of Cs⁺ failed to alter the isometric contractile force significantly. NH₄ ⁺ and K⁺ caused a transient positive inotropic effect which was partially blocked by propranolol. The positive inotropic response to K⁺ was followed by a negative inotropic response. Rb⁺ produced a sustained, dose-dependent inotropic response reaching a plateau at 1–2 min, whereas Tl⁺ produced a dose-dependent positive inotropic effect which developed slowly over a 30-min period. The positive inotropic effects produced by Rb⁺ and Tl⁺ were insensitive to propranolol pretreatment. Concentrations of Tl⁺ and cardiac glycosides which produce similar inotropic effects appear to cause the same degree of Na⁺-pump inhibition. The onset of the positive inotropic response to Rb⁺ or Tl⁺ was not dependent on the number of contractions which is in contrast to the cardiac glycoside-induced inotropic response. Substitution of 20 mM LiCl for an equimolar amount of NaCl in Krebs-Henseleit solution produced a significantly greater inotropic response than that observed when sucrose was substituted for NaCl. It appears that, among monovalent cations, only sodium pump inhibitors produce a sustained positive inotropic response.     

Analysis of the hyperpolarizing effects of forskolin in guinea-pig atrial heart muscle

Summary The effects of forskolin on action potential configuration and on both uptake and efflux of ⁸⁶Rb⁺ were studied in guinea-pig left atria. The action potential was prolonged by forskolin in the plateau range but shortened at the end of repolarization; maximal upstroke velocity and amplitude of slow response potentials were enhanced. In partially depolarized preparations, the resting potential was increased by forskolin; this effect was not prevented by atropine 1 mol/1. Forskolin augmented the rate constant of ⁸⁶Rb⁺ efflux in beating and in resting preparations. The uptake of ⁸⁶Rb^s+ was enhanced by forskolin in resting preparations. It is concluded that forskolin stimulates the Na⁺, K⁺ -pump and activates a background potassium conductance. Both effects may account for the shortening effect of the drug on the action potential and the increase in resting potential seen in partially depolarized preparations. Send offprint requests to H. Nawrath     

Molecular and functional characterization of the human platelet Na(+) /Ca(2+) exchangers

BACKGROUND AND PURPOSE The Na(+) /Ca(2+) exchanger is a bi-directional transporter that plays an important role in maintaining the concentration of cytosolic Ca(2+) ([Ca(2+) ](i) ) of quiescent platelets and increasing it during activation with some, but not all, agonists. There are two classes of Na(+) /Ca(2+) exchangers: K(+) -independent Na(+) /Ca(2+) exchanger (NCX) and K(+) -dependent Na(+) /Ca(2+) exchanger (NCKX). Platelets have previously been shown to express NCKX1. However, initial studies from our laboratory suggest that NCX may also play a role in platelet activation. The objective of this study was to determine if the human platelet expresses functional NCXs. EXPERIMENTAL APPROACH RT-PCR, DNA sequencing and Western blot analysis were utilized to characterize the human platelet Na(+) /Ca(2+) exchangers. Their function during quiescence and collagen-induced activation was determined by measuring [Ca(2+) ](i) with calcium-green/fura-red in response to: changes in the Na(+) and K(+) gradient, NCX pharmacological inhibitors (CBDMB, KB-R7943 and SEA0400) and antibodies specific to extracellular epitopes of the exchangers. KEY RESULTS Human platelets express NCX1.3, NCX3.2 and NCX3.4. The NCXs operate in the Ca(2+) efflux mode in resting platelets and also during their activation with thrombin but not collagen. Collagen-induced increase in [Ca(2+) ](i) was reduced with the pharmacological inhibitors of NCX (CBDMB, KB-R7943 or SEA0400), anti-NCX1 and anti-NCX3. In contrast, anti-NCKX1 enhanced the collagen-induced increase in [Ca(2+) ](i) . CONCLUSIONS AND IMPLICATIONS Human platelets express K(+) -independent Na(+) /Ca(2+) exchangers NCX1.3, NCX3.2 and NCX3.4. During collagen activation, NCX1 and NCX3 transiently reverse to promote Ca(2+) influx, whereas NCKX1 continues to operate in the Ca(2+) efflux mode to reduce [Ca(2+) ](i) .     

Specific activity and sensitivity to strophanthin of the Na+ + K+-activated ATPase in rats and guinea-pigs with hypoadrenalism

Summary In adrenalectomised rats and in guinea-pigs pretreated with metyrapone the specific activity of the Na⁺ + K⁺-stimulated ATPase of heart and kidney is significantly diminished, whereas the activity of the Mg⁺⁺-ATPase remains unchanged. The specific activity of the Na⁺ + K⁺-stimulated ATPase from brain tissue is not influenced by either adrenalectomy or by treatment with metyrapone.The sensitivity of the Na⁺ + K⁺-stimulated ATPase of heart, brain and kidney to k-strophanthin remains unchanged by adrenalectomy or by treatment with metyrapone.Supported by the Deutsche Forschungsgemeinschaft (SFB 30, Kardiologie).     

The effect of diuretics of Na+-K+-ATPase and c-AMP levels in toad bladder epithelial cells

Summary Na⁺-K⁺-ATPase was inhibited by 1x10^–4 M ethacrynic acid and mercuderamide, and by 1x10^–3 M hydrochlorothiazide and furosemide. A modification of Gilman's (1970) protein displacement assay has been used to measure c-AMP levels in toad bladder epithelial cells. Vasopressin (50 mU/ml) caused c-AMP levels to rise from 4.27 to 9.27 pmol/mg protein. Ethacrynic acid had no effect on cellular c-AMP levels after 10 min exposure to the drug, but at 90 min caused a reduction of both basal and vasopressin stimulated levels. Furosemide caused an apparent rise in c-AMP levels, dilution ratio measurements indicated interference by this drug in the assay procedure, mecuderamide also caused substantial interference with the c-AMP assay. Hydrochlorothiazide had no effect on basal or hormone stimulated levels of c-AMP.It was concluded that the inhibition of sodium transport produced by ethacrynic acid in toad bladder is probably due to inhibition of adenylate cyclase, an effect not shared by other diuretics.     

Activation of K+ channels and Na+/K+ ATPase prevents aortic endothelial dysfunction in 7-day lead-treated rats

Seven day exposure to a low concentration of lead acetate increases nitric oxide bioavailability suggesting a putative role of K⁺ channels affecting vascular reactivity. This could be an adaptive mechanism at the initial stages of toxicity from lead exposure due to oxidative stress. We evaluated whether lead alters the participation of K⁺ channels and Na⁺/K⁺-ATPase (NKA) on vascular function. Wistar rats were treated with lead (1st dose 4 μg/100 g, subsequent doses 0.05 μg/100 g, im, 7 days) or vehicle. Lead treatment reduced the contractile response of aortic rings to phenylephrine (PHE) without changing the vasodilator response to acetylcholine (ACh) or sodium nitroprusside (SNP). Furthermore, this treatment increased basal O₂⁻ production, and apocynin (0.3 μM), superoxide dismutase (150 U/mL) and catalase (1000 U/mL) reduced the response to PHE only in the treated group. Lead also increased aortic functional NKA activity evaluated by K⁺-induced relaxation curves. Ouabain (100 μM) plus L-NAME (100 μM), aminoguanidine (50 μM) or tetraethylammonium (TEA, 2 mM) reduced the K⁺-induced relaxation only in lead-treated rats. When aortic rings were precontracted with KCl (60 mM/L) or preincubated with TEA (2 mM), 4-aminopyridine (4-AP, 5 mM), iberiotoxin (IbTX, 30 nM), apamin (0.5 μM) or charybdotoxin (0.1 μM), the ACh-induced relaxation was more reduced in the lead-treated rats. Additionally, 4-AP and IbTX reduced the relaxation elicited by SNP more in the lead-treated rats. Results suggest that lead treatment promoted NKA and K⁺ channels activation and these effects might contribute to the preservation of aortic endothelial function against oxidative stress.     

Acute and chronic effects of lithium in therapeutically relevant concentrations on potassium uptake into astrocytes

Potassium uptake into astrocytes in primary cultures was measured by the aid of ⁴²K. Acute application of lithium in concentrations of 1 and 5 mM, but not 0.5 und 0.25 mM, exerted a significant inhibition of the potassium uptake rates. This effect is due to a partial impairment of the ouabain-sensitive part of the uptake into the cells caused by a lithium interaction with the extracellular K⁺-activated site of the Na⁺, K⁺-ATPase. After 14 days of exposure of the astrocytes to 1 mM lithium, the potassium uptake remained lower in the presence of lithium than in its absence. However, the cells had adjusted to the chronic presence of lithium by increasing their potassium uptake to such an extent that, during the exposure to 1 mM lithium, it was indistiguishable from that in cultures from the same batches grown in the absence of lithium and measured in the absence of this compound. The interference by lithium with potassium uptake into astrocytes may well be related to the inhibition of potassium clearance in the CNS described in the literature.     

Role of Na+-K+ ATPase enzyme in vascular response of goat ruminal artery

Objective:

To study the role of Na⁺, K⁺- ATPase enzyme in the vascular response of goat ruminal artery.

Materials and Methods:

Ruminal artery was obtained in chilled aerated modified Krebs-Henseleit solution (KHS) from a local slaughterhouse and transported in ice for further processing. The endothelium intact arterial ring was mounted in a thermostatically controlled (37 ± 0.5°C) organ bath containing 20 ml of modified KHS (pH 7.4) bubbled with oxygen (95%) and CO₂ (5%) under 2g tension. An equilibration of 90 min was allowed before addition of drugs into the bath. The responses were recorded isometrically in an automatic organ bath connected to PowerLab data acquisition system. In order to examine intact functional endothelium, ACh (10 μM) was added on the 5-HT (1.0 μM) - induced sustained contractile response. Similarly, functional characterization of Na⁺, K⁺-ATPase activity was done by K⁺-induced relaxation (10 μM-10 mM) in the absence and presence of ouabain (0.1 μM/ 0.1 mM), digoxin (0.1 μM) and barium (30 μM).

Results:

ACh (10⁻⁵ M) did not produce any relaxing effect on 5-HT-induced sustained contractile response suggesting that vascular endothelium has no significant influence on the activation of sodium pump by extracellular K⁺ in ruminal artery. Low concentration of Ba²⁺ (30 μM) (IC₅₀: 0.479 mM) inhibited K⁺-induced relaxation suggesting K_ir (inward rectifier) channel in part had role in K⁺-induced vasodilatation in ruminal artery. Vasorelaxant effect of KCl (10 μM-10 mM) in K⁺-free medium is also blocked by ouabain (0.1 μM and 0.1 mM) (IC₅₀:0.398 mM and IC₃₅: 1.36 mM), but not by digoxin (0.1 μM) (IC₅₀ 0.234 mM) suggesting that ouabain sensitive Na⁺, K⁺-ATPase isoform is present in the ruminal artery.

Conclusion:

In the goat ruminal artery functional regulation of sodium pump is partly mediated by K⁺ channel and ouabain sensitive Na⁺, K⁺ ATPase.     

Biochemical basis for the low sensitivity of the rat heart to digitalis

Summary The concentration of cardiac glycosides to produce positive inotropic effects in the rat heart is markedly higher than that in other species. Such a low digitalis sensitivity of the rat heat is attributed to the low affinity of cardiac Na⁺, K⁺-ATPase for digitalis in this species. In the present study the biochemical cause which is responsible for the formation of the unstable complex between the glycosides and Na⁺, K⁺-ATPase or positive inotropic, receptor in the rat heart was examined using Na⁺, K⁺-ATPase preparations obtained from rat hearts, guinea-pig hearts and rat brains as well as isolated, electrically stimulated atrial preparations obtained from these animals. Monensin, which alters transmembrane Na⁺ movements without interacting with the cardiotonic sites on Na⁺, K⁺-ATPase, had equivalent potencies in guinea-pig and rat hearts. Cassaine, which lacks a lactone ring but interacts with cardiotonic sites on Na⁺, K⁺-ATPase, increased the force of contraction in guinea-pig hearts at low, but in rat hearts only at high, concentrations. AY-22,241 (Actodigin) and prednisolone-3,20-bisguanylhydrazone (PBGH) bind to cardiotonic sites on Na⁺, K⁺-ATPase and had a similar spectrum as cassaine in these two species. Actodigin has an altered lactone ring resulting in a marked reduction of the inotropic potency, and PBGH is devoid of this structure. With the latter agent, the rabbit was as insensitive as the rat, although both rabbit and guinea-pig are equally sensitive to digitalis. K⁺ delayed the development of the positive inotropic action of ouabain with a minimal effect on the plateau response in guinea-pig hearts. In rat hearts, however, K⁺ markedly lowered the plateau response without affecting the time course of the response. These results indicate that the low sensitivity of the rat heart to digitalis is due to a difference in the glycoside binding sites on Na⁺, K⁺-ATPase; but the difference cannot be explained by the lack of a lactone ring complementary binding sites. The difference seems to result from the absence of lipid barrier which regulates the rate of release of cardiac glycosides from their binding sites on Na⁺, K⁺-ATPase.This work was supported by U.S. Public Health Service grant, HL-16052 and by the Michigan Heart Association     

Comparative studies of the influence of various K+ concentration on the action of K-strophthidin, digitoxin and strophanthidin-3-bromoacetate on papillary muscle and on membrane-ATPSA of guinea pig hearts

The influence of increasing K⁺ concentrations (5, 16, and 50 mM) on the effects of different cardenolides — digitoxin (DIG), k-strophanthidin (STR) and strophanthidin-3-bromoacetate (SBA) — on the contractile force of isolated electrically stimulated papillary muscles and on the activity of the Na⁺, K⁺-activated ATPase of guinea pig hearts was studied under comparable experimental conditions.