Desensitization to histamine in the absence of external Ca++ in the guinea-pig taenia caecum

Short-term desensitization of the contractile response of the guinea-pig taenia caecum to histamine was tested in the absence of Ca++. Desensitization was monitored both by the fall of histamine response and by the decrease of irreversible blockade by phenoxybenzamine. In Ca++-free solution with 0.2 mM EGTA, desensitization occurred as in normal physiological solution containing Ca++.     

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

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.     

Enzymatic characteristics of the guanylate cyclase of KB cells: their change as a function of the development of the cultures]

We have localized 71% of the guanylate cyclase activity in the (G X 105,000) supernatent fraction of broken KB cells. The reaction follows Michaelis-Menten kinetics, the apparent Km for GTP is 0,5 mM, as long as GTP is lower than a limited concentration, then activity is inhibited. The ion Mn++ is an absolutely required activator, it does not change enzyme-substrate affinity. The enzyme shows several types of binding sites of Mn++. Guanylate cyclase, studied over a period of development of culture, shows, in KB cells without cell contact, an activity higher than that observed in confluent cells. This is not due to the fact of a change in enzyme-substrate affinity but to a modification of Mn++ influence.     

Effect of platelet activating factor on guinea-pig papillary muscle

Platelet activating factor (PAF) induces a biphasic effect on guinea-pig papillary muscle: 1. a transient positive inotropic effect preceded by an increase in action potential duration (APD); 2. a marked negative effect on inotropism and on APD. Since Ca++ slow action potentials were initially enhanced by PAF and then markedly depressed, it is suggested that PAF specifically interferes with the Ca++ slow channel.     

Modulation of 3-hydroxy-3-methyl glutaryl CoA reductase by 2,3-diphosphoglyceric acid

Rat liver microsomal 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase was activated by 50% at a concentration of 0.4 mM 2,3-diphosphoglyceric acid (DPG) and by 11-fold at 10 mM DPG. DPG also prevented the inactivation of HMG-CoA reductase by ATP and Mg++. Rat liver microsomal HMG-CoA reductase prepared in the presence of 1 mM DPG was significantly more active than when prepared in the absence of DPG. Activation of the enzyme by DPG and protection of the enzyme against inhibition by ATP and Mg++ by DPG were also observed with solubilized HMG-CoA reductase.     

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.     

Further support for the postsynaptic action of substance P and its blockade with baclofen in neurons of the guinea-pig hypothalamus in vitro

Effects of substance P on neurons of the guinea-pig hypothalamus in vitro and antagonism between substance P and baclofen were investigated. Substance P increased the firing rate of neurons in the medium containing 0 mM Ca2+ and 12 mM Mg2+. The excitatory action of substance P was antagonized by a low dose of baclofen whereas that of acetylcholine was not antagonized even by much higher doses of baclofen.     

Oscillatory after-potentials and triggered-automaticity in mammalian ventricular muscle fibres at high resting potentials.

Oscillatory after-potentials and triggered-automaticity were observed in dog ventricular muscle fibres when the fibres were exposed to K+-free,high-Ca++-solutions after K+-free,Ca++-free perfusion. They appeared at membrane potentials more negative than--60 m V.     

Extracellular calcium ions modify the effects of Anemonia sulcata toxin (ATX II) in guinea-pig papillary muscles

In isolated guinea-pig papillary muscle ATX II prolonged the action potential duration to a lesser extent at high extracellular Ca++-concentrations. This is interpreted as an interference of Ca++ with ATX II-binding sites.     

The response of Ca-mediated action potentials and contractile activity in mammalian ventricular myocardium towards alkalosis.

Alkalosis (pH 7.8) produced by reduction of CO2 concentration augmented both upstroke velocity of Ca action potentials and isometric contractile force of mammalian heart muscle. If the increase of pH to 7.8 was achieved by a raise of HCO3 concentration (with simultaneous reduction of CO2 concentration), the positive inotropic response was not accompanied by an augmented Ca current. Obviously, the well-known positive inotropic effect of alkalosis does not only depend upon the enhancement of transmembrane Ca influx during excitation, but can be mediated alone by affecting intracellular Ca movements as well.     

Evolution in time of transmitter release at frog neuromuscular junction]

It has previously been shown that the unitary quantal end-plate potentials observed at synapses blocked by low Ca++ high Mg++ ringer, belong to distinct clusters according to their amplitude, time to peak and latency characteristics. These clusters correspond probably to distinct releasing units dispersed along the presynaptic terminal branches. The distribution versus time of the occurence of unitary potentials belonging to one latency cluster has been studied over long lasting evoked nerve activity (stimulation frequencies: 1 to 10 Hz). It was observed that transmitter release at one releasing site is statistically periodic with emitting periods separated by rest periods. At a given end-plate the period of emitting activity seems to be independent from one emitting site to another.     

Calcium transients in a crustacean motoneuron soma: Detection with arsenazo III

Summary Long lasting action potentials can be triggered in crayfish giant motor neurons by a short depolarizing pulse under different conditions. The concomitant increase in absorbance of the Ca indicator arsenazo III preloaded into the soma, confirms observations suggesting that these potential changes are related to a Ca inward current through the some membrane.     

Electrophysiological actions of chlorimipramine on guinea-pig ventricular fibres.

Chlorimipramine (CMI, 1 x 10(-5)M to 7 X 10(-5)M) decreased the amplitude, overshoot and rate of rise of ventricular action potentials and abolished the Ca-mediated action potentials elicited in guinea-pig papillary muscles. These results indicates that CMI inhibits the rise in sodium and calcium conductances during the cardiac action potential.     

Enzymatic characteristics of the catalytic subunit of the protein kinase complex of human lymphocytes]

In earlier reports we have shown the existence in human lymphocytes homogenate, of a cyclic-AMP dependent protein-kinase activity. We demonstrate by affinity chromatography that two subunits display respectively cyclic-AMP binding and phosphorylating properties. Divalent cations such as Ca++, Mg++ or Mn++ are required for enzymatic activity. ATP which is an obligatory cosubstrate acts as an inhibitor when its concentration is higher than 10(-6)M.     

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.     

The response of Ca-mediated action potentials and contractile activity in mammalian ventricular myocardium towards alkalosis

Summary Alkalosis (pH 7.8) produced by reduction of CO₂ concentration augmented both upstroke velocity of Ca action potentials and isometric contractile force of mammalian heart muscle. If the increase of pH to 7.8 was achieved by a raise of HCO₃ concentration (with simultaneous reduction of CO₂ concentration), the positive inotropic response was not accompanied by an augmented Ca current. Obviously, the well-known positive inotropic effect of alkalosis does not only depend upon the enhancement of transmembrane Ca influx during excitation, but can be mediated alone by affecting intracellular Ca movements as well.     

The effects of manganese and barium on the cardiac pacemaker current, if, in rabbit sino-atrial node myocytes

The isolation of ionic fluxes contributing to electric currents through cell membranes often requires block of other undesired components which can be achieved, among others, by divalent cations. Mn2+ and Ba2+ are often used, for example, to block Ca and K currents. Here we have investigated the effects of these two cations on the properties of the hyperpolarization-activated pacemaker current if, in rabbit sino-atrial node myocytes, as obtained by voltage clamp analysis. We find that 2 mM Mn2+ shifts the if activation curve by 3.2 +/- 0.3 mV towards more positive values. However, when 1 mM Ba2+ is also added, the positive shift is more than halved (1.3 +/- 0.2 mV). We find, too, that in the absence of blocking cations the ACh-induced if inhibition is slightly higher than in their presence. These results indicate that the alteration of if kinetic properties by Ba2+ plus Mn(2+)-containing solutions is minimal.     

Zinc antagonizes the effect of botulinum type A toxin at the mouse neuromuscular junction

Zn2+ (10-100 microM) elevated the frequency of miniature end-plate potentials (MEPPs) in the mouse diaphragm. The effect did not depend on external Ca2+. Botulinum type A toxin (BTXA, 50 ng/ml) abolished MEPPs almost completely within 30 min. Zn2+ (100 microM) restored MEPPs and increased their frequency after they had been abolished by BTXA in Ca2+ -free solutions. The antagonistic effect of Zn2+ in the Ca2+ -free solution was reduced by exposing the diaphragm to the toxin in the Ca2+ -free solutions containing high K+. Thus, the action of BTXA is probably enhanced by depolarization of the motor nerve terminals.     

Electrophysiological actions of chlorimipramine on guinea-pig ventricular fibres

Summary Chlorimipramine (CMI, 1×10^–5M to 7×10^–5M) decreased the amplitude, overshoot and rate of rise of ventricular action potentials and abolished the Ca-mediated action potentials elicited in guinea-pig papillary muscles. These results indicates that CMI inhibits the rise in sodium and calcium conductances during the cardiac action potential.     

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

Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10(6) cells/ml in a buffered salt solution and incubated for 1 h at 37 degrees C in 300 microliter volumes in the absence or presence (9 X 10(-4) M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8 X 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 greater than 0.05). It is suggested that D-galactosamine does not require extracellular Ca++ for the release of histamine from the rat mast cell.