Specific inhibition of a calcium dependent activation of brain cyclic AMP phosphodiesterase activity by vinblastine.

Vinblastine selectively inhibits the activation of brain cyclic AMP phosphodiesterase activity by Ca++-protein activator (50% inhibition by 2 x 10(-5) M). This inhibitory effect was reversed by excessive amounts of the activator, whereas large quantities of Ca++ caused only a slight suppression of the vinblastine effect. This result of vinblastine suggests a new site of its action and also suggests the possible role of protein activator, phosphodiesterase proteins or cyclic nucleotides in the previously known effects of vinblastine in vivo and in vitro.     

Effects of propafenone on TEA-induced action potentials in vascular smooth muscle of canine coronary arteries

Summary The Ca⁺⁺-dependent, TEA-induced action potential is blocked by propafenone in a dose-dependent manner. Such results suggests that in coronary arterial smooth muscle one mechanism of action of propafenone is to inhibit Ca⁺⁺ inward current.This research was supported by a grant from the Tennessee Affiliate of the American Heart Association and VA grant No. 1A (74)111-430100). The authors wish to thank Dr Gerhard Pfennigsdorf for supplying us with propafenone. We also wish to thank Ms Penny Williams for her technical assistance.     

Possible regulation of membrane-associated cyclic AMP phosphodiesterase in rat cerebral cortex by lipids

Summary Cyclic AMP phosphodiesterase (PDE) in membrane fraction from rat cerebral cortex was activated by Triton X-100, and treatment at alkaline pH and with phospholipase C. These results suggest that membrane PDE exists in a latent form and is influenced by microenvironmental changes within the membrane. Furthermore, the PDE, unlike soluble enzyme, is not influenced by a protein activator and Ca⁺⁺.     

Phosphorylation of cockroach antennal polypeptides: effects of second messengers and pheromonal blend

In insect antennal extracts, Schleicher et al.¹ showed that protein kinase C (PKC) inhibitors abolish the transience of pheromone-induced rapid inositol trisphosphate responses, which suggests that pheromonal signals act on phosphorylation of specific proteins. To confirm this hypothesis, we studied the effects of second messengers and a pheromonal blend on phosphorylation of antennal proteins in the cockroachPeriplaneta americana. Proteins from adult male antennae were phosphorylated in vitro in the presence of [³²P] triphosphate, then separated by SDS-polyacrylamide gel electrophoresis. Numerous phosphopolypeptides were visualized. The presence of Ca⁺⁺/calmodulin in the incubation medium resulted in increased phosphorylation of polypeptides with molecular weights of 38, 48, 51, 54 and 58 kDa. Stimulation of PKC by addition of Ca⁺⁺ phosphatidylserine (PS)/phorbol myristate acetate (PMA) resulted in the appearance of three phosphopolypeptides of 36, 70 and 120 kDa. In the presence of cyclic adenosine monophosphate, two new major polypeptides of 46 and 42 kDa appeared; the latter polypeptide also appeared in the presence of cyclic guanosine monophosphate. Comparison with polypeptide composition of tissue from the cerci, leg, brain and fat body showed that the 36 and 48 kDa polypeptides were specific to antennae, whereas the 120 kDa polypeptide was also present in the adult brain. When antennae are subjected to pheromonal stimulation for 16 seconds prior to homogenization, in vitro phosphorylation of the 120, 70, 64 and 38 kDa polypeptides was inhibited, whereas phosphorylation of the 58, 54, 51 and 48 kDa polypeptides was strongly stimulated. It is noteworthy that a 107 kDa polypeptide was observed only after pheromonal stimulation by Ca⁺⁺/PS/PMA. Our findings suggest that Ca⁺⁺-and PKC-dependent protein phosphorylation systems play an important role in the transduction of pheromonal signals in antennae of male cockroachP. americana. We speculate that specific phosphoproteins may modulate sensitivity and signal amplification during the olfactory transduction process.     

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.     

Effect of heat treatment on the ATPase activity of various sarcoplasmic reticulum preparations

Summary Ca²⁺-stimulated ATPase activity of sarcoplasmic reticulum (SR) preparations is activated after a short period of preincubation at temperatures between 40 and 45°C, but for temperatures higher than 48°C pronounced denaturation is observed. Heat denaturation is decreased if Mg²⁺ or K⁺ are present during heat treatment.Acknowledgments. This work was supported by research grants from Instituto de Alta Cultura (Grant No. CB/2) and the Calouste Gulbenkian Foundation. We are grateful to Drs.A. P. Carvalho andV. M. C. Madeira for many discussions and suggestions during the course of this work.     

Hypothyroidism lowers blood pressure,adenylate cyclase and Na+, K+-and K+, Ca++-ATPase activities in normotensive and spontaneously hypertensive rats

Summary Myocardial isoproterenol-stimulated adenylate cyclase, Na⁺, K⁺-ATPase and K⁺, Ca⁺⁺-ATPase activities are elevated in the spontaneously hypertensive rat and can be lowered by methimazole-induced hypothyroidism which also prevents the development of hypertension. Although thyroid hormone levels are similar between untreated SHRs and WKY rats, the thyroid is apparently necessary for the expression of spontaneous hypertension.Acknowledgments. Supported by North Carolina Heart Grant No. 40301 and a grant from Sigma Xi.     

Electrical responses of cardiac muscle in Na-free high-Ca solution

Résumé L'activité électrique de préparations atriales de grenouille soumises à un milieu de Ringer dont tout le Na⁺ est remplacé par du Ca⁺⁺ a été enregistrée à l'aide de microélectrodes. Dans ces conditions, il a été possible d'obtenir des potentiels d'action propagés. Ces réponses sont caractérisées par une très grande inversion de potentiel dont l'amplitude, augmentée par la présence d'adrénaline dans le milieu, peut atteindre 90 mV pour une dépolarisation totale de 185 mV, du fait d'une hyperpolarisation de la membrane. Ces résultats démontrent clairement le rôle du Ca⁺⁺ dans l'activité électrique cardiaque.

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Supported by Public Health Service Grant No. 9 R01-14548-14 from the National Heart and Lung Institute.     

Sensitivity of the developing chick myocardium to the positive inotropic effects of calcium and isoproterenol

Summary The present results show that the sensitivity of the chick myocardium to the positive motropic effect of Ca⁺⁺ decreases during development and that the Ca⁺⁺ concentration of the physiological solution used must be lowered below normal to study the effects of positive inotropic agents in preparations from younger embryos. Isoproterenol elicits positive inotropic responses in 7–9-day embryonic ventricle and in newborn chick atria; however, the 4-day embryonic myocardium is unresponsive to isoproterenol.This work was supported by Grant No. HL-15995 from the National Heart Institute (USPHS).The authors would like to thank Dr.F. E. Shideman for the isolated tissue baths used in these experiments.     

Time course of the distribution of in vivo administered89Sr++ in rat liver subcellular fractions

Riassunto É stata studiata la distribuzione dello⁸⁹Sr⁺⁺ nelle frazioni subcellulari di fegato di ratto. Nei primi secondi dopo l'iniezione, la maggior parte dello⁸⁹Sr⁺⁺ si trova nei mitocondri. In alcuni minuti, la radioattività diminuisce nei mitocondri ed aumenta nel reticolo endoplasmico e nella fase solubile. Lo⁸⁹Sr⁺⁺ viene probabilmente trasferito da una struttura cellulare all'altra; i mitocondri ed il reticolo endoplasmico svolgono ruoli distinti nella deposizione dello Sr⁺⁺ nella cellula epatica.

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This research was supported by the U.S. Public Health Service (Grant No. RO5 TW-00216) and by the Italian National Research Council, with the skilled technical assistance of MissDaniela Ceccarelli.     

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.     

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.     

Gilia in human fetal Schwann cells

Summary Cilia are present in Schwann cells during myelinogenesis which suggest that these cells could migrate by ciliary movement.Supported by U. S. PHS Grant No. 8-3360.Acknowledgment. I thank Drs.R. Allen, R. P. Bunge andR. A. Chambers for their advice and criticisms, and Mr.Alan Sandler for technical assistance.     

Isoprenaline-like effects of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine on mechanical,biochemical and electrophysiological parameters in the mammalian heart

Summary The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) mimicked the effects of isoprenaline on the force of contraction, the cAMP content and the slow Ca⁺⁺ inward current (I_si) in isolated guinea pig papillary muscles. The results support the hypothesis that phosphodiesterase inhibitors and -adrenoceptor agonists exert their positive inotropic effects by increasing I_si via the common mediator cAMP.Acknowledgment. This work was supported by the Deutsche Forschungsgemeinschaft.     

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.     

Effects of drugs and ions on a primitive system of spontaneous contractions in a sponge (Euspongia officinalis)

Résumé ChezEuspongia officinalis, l'acetylcholine et l'adrénaline agissent comme excitateurs des contractions spontanées, localisées et brèves, et favorisent en outre l'extrusion des glycoprotéines contenues dans les cellules contractiles. Na⁺ et Ca⁺⁺ sont nécessaires au fonctionnement des pacemakers.

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This work was supported by the Centre National de la Recherche Scientifique (E.R.A. No. 183). We thank Dr.E. A. Robson for the help of her valuable criticisms.     

Ba++ induced square-shaped potential waves in molluscan neurones

Summary Rhythmic slow square-shaped potential waves lasting 20–50 sec are induced in Aplysia neurones when Ca⁺⁺ is replaced by Ba⁺⁺. During the plateau, the membrane is highly permeable to Ba⁺⁺ ions, whereas in the inter-wave period the membrane resistance increases by 60 times. Analysis with slow current ramps suggests that the membrane properties of Ba-treated neurones and normally burst-generating neurones are similar.     

Correlation of ammonia liberation and calcium deposition by the avian egg and blood ammonia levels in the laying hen

Zusammenfassung Mittels biochemischer Daten wird der Nachweis erbracht, dass im Ovidukt des Huhnes während der Schalenbildung zwar vermehrt NH₄ ⁺ freigesetzt wird, jedoch eine negative Korrelation zwischen NH₄ ⁺-Produktion und Ca⁺⁺-Abscheidung besteht.

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Supported by a training grant from the U.S. Public Health Service No. 5-T1-DE-118. We thank Dr.P. R. Tramell for his help during the initial stage of this investigation.     

Elevated kidney glucosyltransferase activity in genetic prediabetic mice

Summary Glucosyltransferase activity in the renal cortex of genetic diabetic KK mice was significantly increased at 40 days of age when compared to that of Swiss albino and F₁ hybrid mice. This increase in enzyme activity in the absence of glucose intolerance can be regarded as an earlier genetic marker for the diagnosis of diabetic microangiopathy.Acknowledgments. This research was supported in part by the General Research Support Grant No. RR-05398 from the General Research Support Branch, Division of Research Resources, National Institutes of Health, National Institutes of Health Training Grant No. 5-T01-AM-05617-06, Hope for Diabetics Foundation, Inc., and Pfizer Research Laboratories. Dr.R. A. Camerini-Davalos was a Career Scientist of the Health Research Council of the City of New York, and Drs.A. S. Reddi andC.A. Velasco were N. I. H. Trainees in Diabetes Mellitus during part of this study.     

Influence of hypoxia on contractility and calcium uptake in rabbit aorta

Summary The influence of hypoxia on noradrenaline (NA)-induced contractions and⁴⁵Ca uptake has been studied on isolated rabbit aortae. Hypoxia significantly decreased the contractility of aortic strips. NA stimulation resulted in increased or decreased⁴⁵Ca uptake by normoxic or hypoxic specimens, respectively. Relating⁴⁵Ca movement with mechanical activity, the results suggest that decrease in Ca⁺⁺ uptake may be mechanism for hypoxic relaxation of aortic smooth muscle.The author acknowledges facilities at the Wellcome Surgical Research Institute, University of Glasgow, U.K. Much of this work was in collaboration with Drs Sheila Jennett and J.D. Pickard.