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A Bouron

Publications and source records attributed to A Bouron.

26 records · Page 2Linked to original sources
PubMed1993

Two sites of action for LCB29 (idrocilamide) in depressing mechanical tension of rat soleus muscle fibers?

The effects of 50 microM LCB29 (idrocilamide) were tested on depolarization-induced and caffeine contractures of rat soleus muscle fibers. When applied intracellularly by free diffusion in cut-end voltage-clamped fibers, LCB29 decreased tension amplitude by about 25%. The same amount of inhibition by LCB29 was observed on contractures induced by 6 mM caffeine. The drug did not affect the repriming of caffeine contractures, indicating that internal recycling of calcium was not affected. The voltage-dependent inactivation of tension was facilitated by external application of LCB29. This effect was calcium dependent, so that the greater the external calcium concentration, the greater the drug effectiveness. The spontaneous relaxation of K+ contractures was also accelerated by LCB29. It is concluded that LCB29 acts intracellularly by decreasing sarcoplasmic reticulum calcium release and externally by facilitating the voltage-dependent inactivation of the voltage sensor for excitation-contraction coupling.

Animals↗
PubMed1993

Kinetic analysis of the L-type calcium current in enzymatically dissociated ferret ventricular myocytes.

The L-type calcium current (ICa-L) was studied in single ferret ventricular myocytes using whole-cell recording with single patch pipettes. Voltage-clamp experiments were performed at room temperature with internal and external Na(+)- and K(+)-free Tyrode solutions in order to isolate ICa-L. For depolarizing steps eliciting small ICa-L the decay of the current is best described by one exponential. For depolarizing steps eliciting large ICa-L (i.e. between -10 and +30 mV), the decay of the current is best described by the sum of two exponentials with a calcium-dependent fast (Tf) time constant and a voltage-dependent slow (Ts) time constant. Experiments conducted with different external concentration of Ca2+ and Ba2+ suggested that the inactivation and the time course of reactivation of the current after a depolarizing pulse are dependent on calcium ions. This confirms previous observations in heart muscle and reveals the existence of a calcium-dependent regulation process of the L-type calcium current in enzymatically dissociated ventricular myocytes from ferret heart.

Animals↗
PubMed1992

Inhibition of protein kinases in rat pheochromocytoma (PC12) cells promotes morphological differentiation and down-regulates ion channel expression.

We have studied morphological differentiation and ion channel expression in PC12 cells under different culture conditions. Differentiation mediated by nerve growth factor (NGF) was compared with that induced by depletion and inhibition of protein kinases (phorbol ester beta-PMA plus staurosporine). Morphological differentiation was similar under both conditions. However, ion channel densities, studied by means of the patch-clamp technique, were enhanced by NGF and reduced by beta-PMA+staurosporine. Similar changes were also observed for omega-conotoxin-sensitive Ca2+ channels by measuring radioligand binding. The decrease in Ca2+ channel density, after treatment of the cells with beta-PMA+staurosporine, resulted in a reduced increase in the intracellular Ca2+ concentration during K+ depolarization. We conclude that morphological differentiation, but not ion channel expression, can occur during depression of protein kinase activities in PC12 cells.

Alkaloids↗
PubMed1992

The L type calcium current in single hypertrophied cardiomyocytes isolated from the right ventricle of ferret heart.

OBJECTIVE: The aim was to study L type calcium current alterations in relation to the action potential lengthening induced by hypertrophy in isolated cardiomyocytes from the right ventricle of ferret. METHODS: Chronic pulmonary artery constriction was established in adult male ferrets under anaesthesia. Sham operated animals were used as controls. Four to six weeks later the heart was excised and treated with a mixed collagenase-elastase solution to isolate the right ventricular myocytes. The calcium current was investigated in control and hypertrophied cells with the whole cell configuration of the patch clamp technique. The validity of the model was tested by analysis of the structural and passive electrical characteristics of the cells, which were enzymatically isolated from right ventricles previously overloaded (4 to 6 weeks) by clipping the pulmonary artery. RESULTS: Isolated cells from right ventricles submitted to a chronic pressure overload had well preserved cellular integrity suggesting the absence of myocardial failure. This compensated form of hypertrophy was characterised by a dilated transverse tubular system, which could explain the increased membrane capacity. Such cells developed a prolonged action potential with a less pronounced fast repolarisation phase inducing a higher plateau phase. When studied in physiological Tyrode solution the density and kinetics of the L type calcium current were not apparently modified, but a significant decrease in density was unmasked when sodium and potassium currents were suppressed by external and internal substitution of sodium and potassium by tetraethyl ammonium. CONCLUSIONS: The decrease in L type calcium current cannot be involved in the lengthening of action potential observed on hypertrophied myocytes, but it could account for the depressed contractile activity. A noticeable decrease of the transient outward current is suggested to explain the action potential alterations.

Action Potentials↗
PubMed1991

Possible involvement of a chloride conductance in the transient outward current of whole-cell voltage-clamped ferret ventricular myocytes.

The transient outward current was studied, using the whole-cell patch-clamp technique, in isolated ventricular cells from the ferret heart. In the presence of 4-aminopyridine and cadmium chloride which respectively blocked the Ca-insensitive and the Ca-dependent outward currents, a residual transient outward current was observed in about 30% of the cells tested. This current was suppressed in external hypochloride solution, completely inhibited by SITS (3 mM) and reversed at the equilibrium potential for chloride ions. This suggests the presence of a chloride permeability which could contribute to the repolarization phase of the cardiac action potential.

Action Potentials↗
PubMed1990

An efficient isolation procedure of Ca-tolerant ventricular myocytes from ferret heart for applications in electrophysiological studies.

A simple procedure which provides a large yield of isolated ferret ventricular myocytes is described. The enzymatic dissociation was performed by perfusion of the whole heart with the "Langendorff method" at 37 degrees C, without an incubation period. Special attention was given to the period of perfusion with Ca-free or low-calcium containing solutions and to the proportion of both collagenase and elastase used. The viability and calcium tolerance of the isolated cells were tested by ultrastructural and electrophysiological studies. Photo-microscopy showed that 60 to 80% of the isolated cells had an elongated shape (18 microns in diameter, 150 microns in length) and did not beat spontaneously in normal Tyrode solution. The morphological and ultrastructural integrity of these cells was shown in SEM by their smooth surface with regularly spaced T-tubule openings and in TEM by the regular distribution of the transverse tubular system, mitochondrium and sarcomeres. Using the whole-cell patch-clamp technique, they had a resting membrane potential of -72 mV, two types ("Purkinje like" and "ventricular like") of action potentials could be elicited and they were correctly affected by well-known modulators of calcium channels. This technique was successfully applied to the rat heart and could be used for heart dissociation of small mammals. It can simultaneously provide isolated cells of different regions of the heart and can be easily and routinely used by any investigator.

Action Potentials↗
PubMed1990

The direct depressant effect of LCB29 (idrocilamide) on mechanical tension of rat soleus muscle fibers.

The effect of LCB29 was tested on twitch characteristics, tetanic tension, and K+ and voltage-clamp contractures of rat soleus muscle fibers. In concentrations ranging from 10(-6) to 5 x 10(-4) M, LCB29 simultaneously inhibited the twitch amplitude, the maximum rate of tension development, and the maximum rate of relaxation. In concentrations ranging from 10(-5) to 10(-4) M, tetanic tension (100 Hz, 1 s) was inhibited by the same amount. The effect of 5 x 10(-5) M LCB29 was studied on K+ contractures and contractures induced, under voltage-clamp conditions, by long-lasting depolarizations. Its effect was significantly stronger than those on twitch and tetanic tension. In addition, LCB29 had a dual effect on strength--duration curves for mechanical threshold. It increased both the rheobasic potential and the steepness of the curve. It is concluded that LCB29 exerts a direct myorelaxant effect on rat soleus muscle; two sites of action are probably involved.

Animals↗