The effect of nifedipine and verapamil on KC1-induced rhythmic contractions of guinea pig ureter in vitro

Addition of KC1 (40 mM) produced rhythmic contractions of guinea-pig ureters in vitro which were unaffected by phentolamine, atropine or tetrodotoxin. KC1 failed to elicit rhythmic contractions of ureters incubated in a Krebs solution with no added Ca++; in these conditions the addition of CaC12 in concentrations of 1.5 mM, or higher, produced rhythmic contractions whose frequency, but not amplitude, was proportional to CaC12 concentration in the bathing medium. EDTA reduced the frequency of KC1-induced rhythmic contractions without affecting their amplitude. Nifedipine and verapamil reduced both the frequency and the amplitude of KC1-induced rhythmic contraction; verapamil was more effective than nifedipine in reducing their amplitude. Urethane reduced the amplitude without significantly affecting the frequency of KC1-induced rhythmic contractions. An increase in the extracellular Ca++ concentration reverted the suppressive effect of all drugs under study. These results suggest that an influx of Ca++ from the extracellular space is responsible for the initiation of KC1-induced rhythmic contractions and is involved in the mechanism(s) which regulates their frequency, but that a separate mechanism regulates their amplitude.     

An analysis of the effects of urethane on cardiovascular responsiveness to catecholamines in terms of its interference with Ca++ mobilization from both intra and extracellular pools

Summary Urethane (1×10^–2–1×10^–1 M) reduced, in a concentration-dependent manner, both intra and extracellular Ca⁺⁺ dependent noradrenaline-induced contractions of perfused rabbit ear artery as well as the tonic contractions produced by perfusion with high K⁺ solution. However, a quantitative analysis of the data indicated that for urethane concentrations similar to those found in plasma during anesthesia urethane antagonism is confined to noradrenaline-induced contractions which depend upon the mobilization of Ca⁺⁺ from intracellular storage sites. In KCl-contracted arteries, urethane enhanced the relaxant effects of isoprenaline.—Urethane reduced the amplitude of contractions of spontaneously beating guinea-pig right atrium at concentrations which have only a limited effect on frequency. In addition, it decreased in a concentration-dependent manner the amplitude of isoprenaline-activated electrically driven, and K⁺ depolarized guinea-pig right ventricular strips. Urethane had no effect on the chrono and inotropic actions of isoprenaline on cardiac preparations. In in vivo experiments the chronotropic response to low doses of isoprenaline was significantly higher in urethane-treated as compared to unanesthetized rats. The higher dose of isoprenaline tested produced a significant fall in systolic blood pressure in urethane-anesthetized rats. A significant correlation exists between the chronotropic response to isoprenaline and resting heart rate values in urethane-anesthetized rats. These results indicate that urethane, at concentrations similar to those found in plasma during anesthesia selectively interferes with mobilization of Ca⁺⁺ from intracellular storage sites. In addition, the interference of urethane anesthesia with the isoprenaline chronotropic effect in vivo cannot be explained by a direct interference of urethane with -adrenoceptors at cardiac level.     

Influence of ethanol on calcium,inositol phospholipids and intracellular signalling mechanisms

Summary Studies have implicated Ca⁺⁺ in the actions of ethanol at many biochemical levels. Calcium as a major intracellular messenger in the central nervous system is involved in many processes, including protein phosphorylation enzyme activation and secretion of hormones and neurotransmitters. The control of intracellular calcium, therefore, represents a major step by which neuronal cells regulate their activities. The present review focuses on three primary areas which influence intracellular calcium levels; voltage-dependent Ca⁺⁺ channels, receptor-mediated inositol phospholipid hydrolysis, and Ca⁺⁺/Mg⁺⁺-ATPase, the high affinity membrane Ca⁺⁺ pump.Current research suggests that a subtype of the voltage-dependent Ca⁺⁺ channel, the dihydropyridine-sensitive Ca⁺⁺ channel, is uniquely sensitive to acute and chronic ethanol treatment. Acute exposure inhibits, while chronic ethanol exposure increases⁴⁵Ca⁺⁺-influx and [³H]dihydropyridine receptor binding sites. In addition, acute and chronic exposure to ethanol inhibits, then increases Ca⁺⁺/Mg⁺⁺-ATPase activity in neuronal membranes. Changes in Ca⁺⁺ channel and Ca⁺⁺/Mg⁺⁺-ATPase activity following chronic ethanol may occur as an adaptation process to increase Ca⁺⁺ availability for intracellular processes. Since receptor-dependent inositol phospholipid hydrolysis is enhanced after chronic ethanol treatment, subsequent activation of protein kinase-C may also be involved in the adaptation process and may indicate increased coupling for receptor-dependent changes in Ca⁺⁺/Mg⁺⁺-ATPase activity.The increased sensitivity of three Ca⁺⁺-dependent processes suggest that adaptation to chronic ethanol exposure may involve coupling of one or more of these processes to receptor-mediated events.     

The affinity of mitochondria for Ca++

Riassunto E' stata studiata l'affinità dei mitocondri di fegato di ratto e di cuore di coniglio per il Ca⁺⁺, nel sistema dipendente da energia. Si sono usati tamponi EGTA-Ca⁺⁺ per mantenere la concentrazione del Ca⁺⁺ costante durante l'esperimento. Si è dimostrato che laK _m apparente per il Ca⁺⁺ non è lontana da 10^–6 M.

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We are indebted to Dr.B. Chance for many stimulating discussions and for facilitating the progress of the work with the generous hospitality in his Institute.     

Effect of trifluoperazine and calcium ions on gregarine gliding

Summary The drug trifluoperazine (TFP) inhibits the gliding motility of gregarine protozoans. This suggests that the Ca⁺⁺ binding protein, calmodulin, plays a role in the motility process. However the presence of extracellular Ca⁺⁺ was not required for gliding to occur.Acknowledgments. We should like to thank L. Cooper for technical assistance, Smith, Kline and French for the gift of trifluoperazine, and Terry Preston for valuable discussion.     

Calcium and magnesium in plant cytokinesis and their antagonism with caffeine

Summary The efficiency of caffeine at different concentration on the induction of binucleate cells in onion root-tip was studied. The drug effect is strongly depressed in the Ca⁺⁺ and/or Mg⁺⁺ presence at half-rate of maximum efficiency (0.04%), about 2 mM). We therefore conclude that both cations must play a role in plant cytokinesis.     

Effects of prostaglandin E1 on45Ca++-incorporation and spike activity in longitudinal smooth muscle of cat jejunum

Summary Prostaglandin E₁ (0.3 M) decreased both the⁴⁵Ca⁺⁺-incorporation and the spike activity in isolated longitudinal smooth muscle preparations of the cat jejunum probably by an inhibition on the Ca⁺⁺ influx.     

Possible role of intestinal alkaline phosphatase activity in thiamine transport

Summary Thiamine deficiency caused a marked decrease of intestinal alkaline phosphatase (al-Pase) activity, but had no effect on the Ca⁺⁺-ATPase activity and Ca⁺⁺-absorption in rats. The al-Pase activity was significantly decreased 1 h after oral administration of ethanol at 0.5 and 2.5 g/kg. In contrast, Mg⁺⁺-, Ca⁺⁺- and (Na⁺+K⁺)-ATPase activities did not change after the administration of ethanol. These findings show that the al-Pase activity, unlike the Ca⁺⁺-ATPase activity, is not related to Ca⁺⁺-absorption. A possible role of al-Pase activity in the active transport of thiamine in the intestine was discussed.     

High K+-induced release of somatostatin from the cortical preparation of rat brain

Summary Release of endogenous somatostatin (SRIF) from the rat cerebral cortical slices incubated in Krebs-bicarbonate buffer was increased from the basal rate of 3.4±0.6% of the total SRIF content in 15 min at [K⁺]_o=5.6 mM, to 13.1±1.6% upon raising the [K⁺]_o to 56.6 mM. The high-K⁺ evoked SRIF release was absent when Ca⁺⁺ in the medium was replaced by Mn⁺⁺. The isolated synaptosomes from rat cerebral cortex contain 13.2±3.1 ng SRIF/mg protein compared to 0.33±0.01 ng/mg protein in the cortical tissue as a whole, suggesting that nerve terminals are the main source of the peptide released upon membrane depolarization.The study was supported by a grant from the Medical Research Council of Canada. Results of this work have been published in part as abstracts: Can. Physiol.9, 45 (1978), and Fedn Proc.37, 665 (1978).The authors are greatly indebted to Dr M. Gotz and the Ayerst Research Laboratories for the most generous supply of the synthetic somatostatin.     

Extracellular space values and intracellular ionic concentrations in the isolated midgut of Philosamia cynthia and Bombyx mori

Summary Total extracellular space values have been determined in the midguts of 2 lepidopteran larvae, Philosamia cynthia and Bombyx mori. The values found are 42% and 45% tissue water respectively. Intracellular concentrations of Na⁺, Ca⁺⁺ and Mg⁺⁺ are very low, while K⁺ concentration is 197.2 mEq/l cell water in Philosamia and 180.9 mEq/l cell water in Bombyx.     

Ambiguous effect of caffeine upon the transmembrane Ca current in mammalian ventricular myocardium

Zusammenfassung Coffein (20 mM) wirkt auf den transmembranären Ca⁺⁺-Strom der Warmblütermyokardfaser biphasisch. Zunächst nimmt der Ca⁺⁺-Strom zu, was auf einer Verbesserung der Ca⁺⁺-Leifähigkeit des langsamen Membrankanals beruht. Nach längerer Einwirkungsdauer von Coffein kommt es dagegen zu einer Abnahme des Ca⁺⁺-Stroms. Als Ursache hierfür wird vor allem eine Verminderung des transmembranären Konzentrationsgradienten für Ca⁺⁺ — infolge eines Anstiegs der freien Ca⁺⁺-Konzentration im Zellinnern — postuliert.     

Evidence for cross bridge slippage in a stretched muscle fibre

Summary Bundles of glycerol-extracted psoas fibres which were contracted by immersion in a saline containing 15 mM MgATP and 12 M free Ca⁺⁺ were subjected to up to 3 stretches (rise time 0.8 msec) each of amplitude 1% L_i at intervals of 10 msec. The elastic tension responses to these stretches were all of comparable size and the peak tensions reached during the stretches were in each case followed by a rapid tension decline almost to the tension values before the stretches. This indicates that stretch-induced detachment and reattachment of cross bridges to the actin filament occurred within 10 msec (slippage).Supported by the Deutsche Forschungsgemeinschaft, grant Ru 154/12. The editorial assistance of Mr D.J. Williams is gratefully acknowledged.     

Influence of bistramide A on the twitch tension in rat atrial heart muscle

Bistramide A, a new toxin isolated from the UrochordateLissoclinum bistratum Sluiter, was applied to rat auricular heart muscle bundles. At a stimulation frequency of 0.2 Hz, the toxin induces a dose-dependent reduction of the stimulated twitch tension force; it decreases and shortens the duration of the plateau and the slow repolarizing phase of the action potential. In the control solution, switching from a stimulation frequency of 0.2 Hz to 1 Hz decreases the force with which a positive potentiation develops either at a maintained high frequency or after switching from 1 Hz to 0.2 Hz. Bistramide A reduces both the force evoked at 1 Hz and the potentiation. The data suggest that Bistramide A blocks Na⁺ conductance; inhibits Ca⁺⁺ channels in a time-and frequency-dependent manner; reduces Na⁺–Ca⁺⁺ exchange activity; but does not modify the ability of the sarcoplasmic reticulum to be refilled although the rate of Ca⁺⁺ accumulation is decreased.     

Zinc ions block the intracellular calcium release induced by caffeine in guinea-pig taenia caeci

Zn²⁺ in low concentrations (0.005–0.1 mM) inhibited the transient contractions in response to caffeine (25 mM) in a dose-dependent manner in smooth muscle of intact guinea-pig taenia caeci. At Zn²⁺ concentrations higher than 0.1 mM, caffeine did not elicit any response. After saponin-treatment of the fibres, which leaves the Ca²⁺ storage sites intact, caffeine contraction was completely inhibited by Zn²⁺ at a relatively low concentration (0.03 mM). However, in Triton-X-100-treated fibres, in which the Ca²⁺ release sites are destroyed, the contraction could be induced in the presence of Zn²⁺ by an increase in Ca²⁺. In conclusion, Zn²⁺ can block the intracellular Ca²⁺ release from caffeine-sensitive release sites in taenia caeci.     

Role of calcium in the histamine release induced by D-galactosamine from rat mast cells

Summary Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10⁶ cells/ml in a buffered salt solution and incubated for 1 h at 37°C in 300 l volumes in the absence or presence (9×10^–4 M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8×10^–4 M) caused (in addition to basal release) a mean ±SEM percent histamine release of 15.7±5.2 in the presence of Ca⁺⁺ and 19±4.9 in the absence of Ca⁺⁺ (p>0.05). It is suggested that D-galactosamine does not require extracellular Ca⁺⁺ for the release of histamine from the rat mast cell.A preliminary analysis of these results was presented at the International Symposium on calcium entry blockers and tissue protection, Rome, 15–16 March 1984.     

Die Abhängigkeit der Ca++-Aufnahme isolierter Mitochondrien des Herzmuskels von der Na+-und K+-Konzentration als mögliche Ursache der inotropen Digitaliswirkung

Summary In isolated mitochondria of heart muscle from rabbits and oxen there is, under suitable conditions, an accumulation of Ca⁺⁺, which is significantly enhanced by elevating the K⁺/Na⁺ quotient of the incubation medium. K-strophanthine (10^–5–10^–7) does not influence the accumulation of Ca⁺⁺ by the mitochondria of heart muscle. Therefore the intracellular increase in exchangeable Ca⁺⁺ observed after digitalis-glycosides could be explained by a decrease of the intracellular K⁺/Na⁺ quotient, which is caused by inhibition of the membrane ATPase and diminishes the capacity for Ca⁺⁺ accumulation in mitochondria.     

The reaction of Ca++ with the inner and outer membrane of mitochondria

Riassunto È stata studiata la distribuzione del transporto attivo del Ca⁺⁺ e dei siti ad alta e bassa affinità per il Ca⁺⁺ tra la membrana esterna e interna dei mitocondri di fegato. La membrana interna trasporta attivamente il Ca⁺⁺, la membrana esterna non è in grado di farlo. Ambedue le membrane posseggono i siti a bassa affinità, mentre i siti ad alta affinità si trovano solo sulla membrana interna.     

Influence de la vagotomie chez la tancheTinea tinca au cours d'une surcharge pluricationique (eau de mer diluée). Variation des cations Na+, K+, Ca++ et de l'intensité de la respiration tissulaire

Summary Respiratory ionic content (Na⁺, K⁺, Ca⁺⁺) of hepatic tissue and ionic excretion measurements were performed in the TenchTinea tinea kept in diluted sea water. After bilateral section of parasympathetic system, the changes of Na/K value are less than in fresh water. The survival and hepatic O₂ uptake were the same. It is suggested than after both vagi section sea water Ca⁺⁺ allowed a best osmoregulation in this species.

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Ce travail a été réalisé avec la collaboration technique de A.Rup.     

Mechanical and biochemical characterization of the contraction elicited by a calcium-independent myosin light chain kinase in chemically skinned smooth muscle

Summary The contraction induced by a Ca²⁺-independent myosin light chain kinase (MLCK-) was characterized in terms of isometric force (F_o), immediate elastic recoil (SE), unloaded shortening velocity (V_us), shortening under a constant load and ATPase activity of chemically skinned smooth muscle preparations. These parameters were compared to those measured in a Ca²⁺-induced contraction to assess the nature of cross bridge interaction in the MLCK-induced contraction. F_o developed in chicken gizzard fibers as well as SE were similar in contractions elicited by either agent. V_us in the contraction induced by MLCK-(0.36 mg/ml) was similar though averaged 39.3±8.9% less than V_us induced by Ca²⁺ (1.6x10^–6M) in the control fibers. Addition of Ca²⁺ (1.6x10^–6M) to a contraction induced by MLCK-resulted in small increases in both F_o and V_us. Shortening under a constant load was similar for both types of contractions. The contraction induced by MLCK-was accompanied by an increased rate of ATP hydrolysis. The MLCK-induced contraction is thus kinetically similar though not identical to a contraction induced by Ca²⁺. We conclude that with respect to actin-myosin interaction, MLCK- and Ca²⁺-induced contractions are similar.     

Magnesium-calcium antagonism in the contraction of arterioles

Zusammenfassung Erhöhung des Mg-Spiegels im Blut hemmt Tonus und Kontraktilität der Widerstandsgefässe im Muskel. Durch Noradrenalin hervorgerufene Verengerung wird gleich stark gehemmt wie die durch Reizung sympathischer Nerven verursachte Kontraktion. Ein Überschuss an Ca⁺⁺ verhindert die blockierende Wirkung des Mg⁺⁺ auf die Kontraktilität, der Tonusverlust der Gefässe wird durch Ca⁺⁺ jedoch nicht rückgängig gemacht. Die antagonistischen Wirkungen von Mg⁺⁺ und Ca⁺⁺ beruhen wahrscheinlich auf deren gegensätzlicher Wirkung auf die elektromechanische Kopplung im Gefässmuskel.

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U.S. Public Health Service International Fellow.