[Traumatic subcutaneous emphysema of the face: the hazard of holding a sneeze]. / L'image du mois. Emphysème sous-cutané traumatique de la face : le danger de retenir un éternuement.

In this period of pandemic, protective measures and social distancing, sneezing might not be well received and trying to suppress a sneeze is tempting. It's not always a good idea. We here report the case of a patient suffering from minor facial trauma. The next day after the accident, while trying to hold back a sneeze, the patient caused sudden swelling of the right cheek, associated with subcutaneous emphysema and disturbances in sensitivity, revealing an unrecognized fracture of the right maxillary sinus. Post-traumatic subcutaneous emphysema of the face, caused by suppressed sneezing, is rarely described.

En cette période de pandémie, de gestes barrières et de distanciation sociale, éternuer peut être mal perçu et chercher à réprimer un éternuement est tentant. Ce n'est pas toujours une bonne idée. Nous rapportons ici le cas d'un patient victime d'un traumatisme facial mineur. Le lendemain de l'accident, en essayant de retenir un éternuement, le patient provoqua un brusque gonflement de la joue droite, associé à un emphysème sous-cutané et à des troubles de la sensibilité, révélant ainsi une fracture méconnue du sinus maxillaire droit. L'emphysème sous-cutané post-traumatique de la face, provoqué par un éternuement réprimé, est rarement décrit.

[Haemophilic ankle arthropathy : case reports and review of the literature]. / Arthropathie hémophilique de la cheville : description de deux cas et revue de la littérature.

Haemophilic arthropathy affects about half of the patients who suffer from haemophilia. Despite the fact that it's one of the main morbidity factors of haemophilia and that the pathophysiology of its mechanism is slowly better understood, its management is still under discussion. The cases of two men (53 and 54 years old) who suffer from ankle haemophilic arthropathy since several years are reported. For both cases, different aspects of the management are investigated, including a medicated and a physiotherapy approach, and an adequate orthotic. Other treatments are available and sometimes used, such as radio- or arthroscopic synovectomy, corticosteroids or platelet-rich plasma in?ltration or visco-supplementation.

L'arthropathie hémophilique affecte environ la moitié des hémophiles. Malgré le fait qu'elle soit un des facteurs principaux de morbidité dans l'hémophilie et que la physiopathologie de ce mécanisme soit de mieux en mieux comprise, sa prise en charge est toujours sujette à discussion. Nous décrivons le cas de deux hommes (âgés de 53 et 54 ans) qui souffrent d'arthropathie hémophilique de cheville depuis plusieurs années. Pour chacun, différents aspects de prise en charge ont été investigués, dont les approches médicamenteuses, rééducatives, et de port d'orthèse adéquate. D'autres prises en charge sont également disponibles et parfois utilisées, comme la radiosynovectomie ou la synovectomie arthroscopique, les in?ltrations par dérivés cortisonés ou de plasma riche en plaquettes (PRP), ou encore une visco-supplémentation.

[Innovations in physical and rehabilitation medicine]. / Innovations en médecine physique et réadaptation.

Over the last decade, Physical and Rehabilitation Medicine (PRM) is a medical specialty that has evolved considerably in the various fields that concern it : from the management of low back pain and lumbosciatalgia or osteoporosis in a multidisciplinary manner, through the use of new technologies in neuro-locomotor rehabilitation and robotisation in amputee patients for example, the development of regenerative medicine and prevention in sports traumatology and, finally, the progress of electrophysiology techniques for the diagnosis of small-fibre neuropathies. These various advances will be discussed in this article.

Au cours de la dernière décennie, la Médecine Physique et Réadaptation (MPR) est une spécialité médicale qui a fortement évolué dans les différents domaines qui la concernent : de la prise en charge des lombalgies et lombosciatalgies ou encore de l'ostéoporose de manière pluridisciplinaire, en passant par l'utilisation des nouvelles technologies en rééducation neuro-locomotrice et de la robotisation en rééducation, chez les patients amputés par exemple, le développement de la médecine à vocation régénérative et la prévention en traumatologie du sport et, enfin, les progrès des techniques d'électrophysiologie pour le diagnostic des neuropathies à petites fibres. Ces différentes avancées seront abordées dans cet article.

Modulation by stimulation rate of basal and cAMP-elevated Ca2+ channel current in guinea pig ventricular cardiomyocytes.

The modulation of L-type Ca2+ current (ICa) by changes in stimulation frequency was investigated in single ventricular cardiomyocytes isolated from guinea pig hearts. Electrical recordings were carried out at 21-25 degrees C and at 33-37 degrees C with the whole-cell patch clamp method, under K(+)-free conditions. A comparison is made between the response to frequency changes for ICa in the basal state and after the application of drugs which elevate the level of adenosine-3',5'-cyclic monophosphate (cAMP) within the cells. Peak basal ICa was reduced with an increase in stimulation rate from 0.5 Hz to 1, 2, 3, 4, or 5 Hz. This frequency-induced reduction of ICa was enhanced by reduced temperature, was unchanged when Na+ or Ba2+ carried the basal Ca2+ channel current, and was greatly enhanced after elevating cAMP levels with forskolin, isoprenaline, or 8-(4-chlorophenylthio)-cyclic AMP. We examined the mechanism of the enhancement of the frequency-induced reduction of ICa by cAMP, and found two conditions which abolished it: (a) application of isoprenaline when Na+ carried the Ca2+ channel current in Ca(2+)-free solution, or (b) application of 3-isobutyl-1-methylxanthine, a broad-spectrum phosphodiesterase inhibitor. It was further shown that an elevation of both ICa and cAMP (induced by isoprenaline), and not an increase of ICa alone (induced by Bay K 8644), is required to produce the extra component of reduction by frequency. It is concluded that Ca2+ entry results in feedback regulation of ICa, through the activation of Ca(2+)-dependent phosphodiesterase(s). This is important in the context of sympathetic stimulation, which produces the companion conditions of an elevated heart rate and increases in cAMP levels and Ca2+ entry.

Calcium channels reconstituted from the skeletal muscle dihydropyridine receptor protein complex and its alpha 1 peptide subunit in lipid bilayers.

In the first part of this study, we show that sDHPR and pDHPR preparations reconstituted into lipid bilayers formed on the tips of patch pipettes exhibit two divalent cation-selective conductance levels of 9 and 20 pS, similar in single-channel conductance to VSCC reported in a variety of intact preparations (see Pelzer et al. and Tsien et al. for review). The larger conductance level is similar to the VSCC identified in intact rat t-tubule membranes and described in sDHPR and pDHPR preparations, and shares many properties in common with activity from L-type VSCC. It is sensitive to augmentation by the DHP agonist (+/-)-BAY K 8644 and cAMP-dependent phosphorylation, and to block by the phenylalkylamine (+/-)-D600 and the inorganic blocker CoCl2. Its open-state probability and open times are increased upon depolarization as expected for a voltage-dependent activation process. Upon depolarization beyond the reversal potential, however, open-state probability and open times decline again. A reasonable way to explain the bell-shaped dependence of open times and open-state probability on membrane potential is to assume voltage-dependent ion-pore interactions that produce closing of the channel at strong negative and positive membrane potentials. By contrast, the smaller conductance level may be similar to the 10.6-pS t-tubule VSCC described by Rosenberg et al. and may best be compared with T-type VSCC. It is largely resistant to augmentation by (+/-)-BAY K 8644 and cAMP-dependent phosphorylation or block by (+/-)-D600, but is sensitive to block by CoCl2. Its open times and open-state probability show a sole dependence on membrane potential where depolarization increases both parameters sigmoidally from close to zero up to a saturating level. Both elementary conductance levels do not exhibit significant inactivation over a wide potential range, which may suggest that skeletal muscle VSCC inactivation is either poorly or not voltage-dependent at all. This possibility seems in agreement with bilayer recordings on reconstituted intact t-tubule membranes and voltage-clamp recordings on intact fibers. It supports the idea that the decline of Ca2+ current in intact skeletal muscle fibers may be due to Ca2+ depletion from the t-tubule system and/or to inactivation induced by Ca2+ release from the sarcoplasmic reticulum. We consistently observe two conductance levels of 9 and 20 pS, either singly, or together in the same bilayer from solubilized DHPR samples and even highly purified DHPR preparations.(ABSTRACT TRUNCATED AT 400 WORDS)

The electrophysiological basis of the bradycardic action of AQA 39 on the sinoatrial node.

The electrophysiological effects of the bradycardic agent AQA 39 (5,6-dimethoxy-2-[3-[[ alpha-(3,4-dimethoxy) phenylethyl] methylamino] propyl] phthalimidine hydrochloride) on small preparations of the S-A node of the rabbit heart were investigated. The following results were obtained: 1. The decrease in the rate of the spontaneous preparation resulted from a lower rate of diastolic depolarization, a slower upstroke and a longer duration of the action potential. Concomitantly, overshoot and maximum diastolic potential were decreased. 2. The drug effect on rate strongly depended on the potential during diastole. AQA 39 acted stronger the more positive the maximum diastolic potential. 3. In voltage-clamp experiments, the membrane potential was held at -40 mV and transiently depolarized by square pulses to -10 mV. At a low pulsing rate (0.005 Hz), the main effect was a reduction of the time-dependent potassium current (Ik); the slow inward current (Isi) was only slightly reduced. However, when the pulsing rate was increased to 1 Hz, a clear reduction of Isi was observed. 4. When the block of Ca channels had reached a steady state during continuous pulsing in the presence of the drug, part of the block could be removed by rest periods, relief of block being dependent on the membrane potential during rest. At a fixed rest period of 45s, relief of block was nearly complete for potentials negative to -55 mV but negligible positive to -35mV. 5. AQA 39 shifted the dose-response curve to ionophoretic application of acetylcholine to higher concentrations, suggesting a competitive action of the drug with acetylcholine at the muscarinic receptor.

Ultraviolet photoalteration of late Na+ current in guinea-pig ventricular myocytes.

UV irradiation has multiple effects on mammalian cells, including modification of ion channel function. The present study was undertaken to investigate the response of membrane currents in guinea-pig ventricular myocytes to the type A (355, 380 nm) irradiation commonly used in Ca(2+) imaging studies. Myocytes configured for whole-cell voltage clamp were generally held at -80 mV, dialyzed with K(+)-, Na(+)-free pipette solution, and bathed with K(+)-free Tyrode's solution at 22 degrees C. During experiments that lasted for approximately 35 min, UVA irradiation caused a progressive increase in slowly-inactivating inward current elicited by 200-ms depolarizations from -80 to -40 mV, but had little effect on background current or on L-type Ca(2+) current. Trials with depolarized holding potential, Ca(2+) channel blockers, and tetrodotoxin (TTX) established that the current induced by irradiation was late (slowly-inactivating) Na(+) current (I(Na)). The amplitude of the late inward current sensitive to 100 microM: TTX was increased by 3.5-fold after 20-30 min of irradiation. UVA modulation of late I(Na) may (i) interfere with imaging studies, and (ii) provide a paradigm for investigation of intracellular factors likely to influence slow inactivation of cardiac I(Na).

Currents through ionic channels in multicellular cardiac tissue and single heart cells.

Ionic channels are elementary excitable elements in the cell membranes of heart and other tissues. They produce and transduce electrical signals. After decades of trouble with quantitative interpretation of voltage-clamp data from multicellular heart tissue, due to its morphological complexness and methodological limitations, cardiac electrophysiologists have developed new techniques for better control of membrane potential and of the ionic and metabolic environment on both sides of the plasma membrane, by the use of single heart cells. Direct recordings of the behavior of single ionic channels have become possible by using the patch-clamp technique, which was developed simultaneously. Biochemists have made excellent progress in purifying and characterizing ionic channel proteins, and there has been initial success in reconstituting some partially purified channels into lipid bilayers, where their function can be studied.

Activity-induced facilitation of L-type Ca2+ current in cardiomyocytes isolated from guinea-pig ventricles.

The facilitation of L-type Ca2+ current (ICa), which sometimes occurs with an increase in stimulation frequency, was investigated in single guinea-pig ventricular cardiomyocytes using whole-cell recording and K(+)-free solutions. With a holding potential of -80 mV, an increase in frequency from 0.5 to 1, 2, 3 or 4 Hz caused either a small or large initial reduction, or a transient enhancement (facilitation) of peak ICa, which developed rapidly and was followed by a reduction of ICa. Reducing the frequency to 0.1 or 0.2 Hz caused a depression of ICa on the first pulse that was followed by a slower increment. Transient facilitation and depression were entirely absent when either Ba2+ or Na+ was used as the charge carrier in Ca(2+)-free solutions. High concentrations of isoprenaline (1-3 microM), forskolin (1-3 microM), or 8-(4-chlorophenylthio)-cAMP (150 microM) suppressed but did not abolish the incidence and size of facilitation; employing a holding potential of -40 mV also suppressed the incidence of ICa facilitation. Lower isoprenaline concentrations (0.1 and 0.3 microM) greatly enhanced the incidence and magnitude of the transient facilitation occurring with an increase in stimulation rate, but did not diminish the magnitude of the ensuing reduction. When facilitation occurred with 0.1 mM EGTA in the dialysate, or the usual 5 mM EGTA with 0.01 microM extracellular isoprenaline, it developed more slowly after an increase in frequency. In the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, an increase in stimulation rate from 0.5 to 1, 2 or 3 Hz sometimes caused a large and sustained facilitation of ICa, which developed over tens of seconds, declined slowly with continued stimulation and was maintained after returning to 0.5 Hz. It is concluded that the levels of intracellular cAMP and Ca2+ mediate the initial sensitivity of ICa to changes in stimulation rate, to the extent that they determine whether or not transient facilitation will occur. Because the reduction of ICa was relatively constant, facilitation dictates the level of steady-state ICa that will be reached at the higher rate. Taken together with the fact that facilitation can be modulated, the results argue for separate mechanisms of facilitation and reduction for ICa. It is suggested that the mechanism of facilitation is partly enzymatic, insofar as sustained facilitation could be a manifestation of a stimulatory Ca(2+)-dependent process, which is normally counteracted by the action of phosphodiesterases.(ABSTRACT TRUNCATED AT 400 WORDS)

Dual action (stimulation, inhibition) of D600 on contractility and calcium channels in guinea-pig and cat heart cells.

1. We examined the effects of D600 (0.2-40 microM, generally 2 microM) on the following (i) developed tension in guinea-pig papillary muscles, (ii) calcium current (Ica) and tension in cat ventricular muscle strands, (iii) Ica in guinea-pig and cat ventricular myocytes, (iv) single Ca2+ channel currents carried by Ba2+ in cell-attached membrane patches of guinea-pig ventricular myocytes, and (v) Ba2+ currents through dihydropyridine (DHP)-binding sites (skeletal muscle) reconstituted into single functional Ca2+ channels in lipid bilayers. 2. In 27 of 140 preparations studied, D600 elicited a transient stimulation that preceded marked inhibition. The stimulation was normally of short duration (less than 5 min) and moderate strength (less than 50% increase). 3. D600 had no effect on the unit conductance of single cardiac Ca2+ channels. Stimulation was characterized by a decrease in the number of records with no openings (blanks) and an increase in the open-state probability of non-blanks (longer open times, shorter closed times). Inhibition began with an increase in the number of blanks and later included a curtailment of open times and a prolongation of closed times. The net effect after 9 min D600 was a 75% reduction in average current amplitude. 4. A similar pattern of changes in channel open and closed times produced enhancement and then depression of time-averaged open-state probability in single reconstituted channels. 5. Single Ca2+ channel current that was stimulated by adrenaline was only slightly depressed after 2 microM-D600 for 30 min. It may be that channel phosphorylation or Gs-protein activation following beta-receptor stimulation reduces channel affinity for D600. 6. Short-lived binding of D600 to a single inhibitory site may enhance association/activation of Gs-protein and thereby cause transient up-regulation prior to increased drug occupancy and inhibition. Alternatively, there may be separate stimulatory and inhibitory sites. One aspect of inhibition, the increased frequency of blanks, is attributed to a stabilization of the inactivated state; the other aspect, changes in fast kinetics, seems to require a different explanation.

Cardiac Ca channel currents at the level of single cells and single channels.

Ca channels in excitable membranes are of great importance for many cellular functions. Modulation of these channels by voltage, neurotransmitters and drugs regulates Ca influx into the cell and thereby alters the functional state of the cell (7). For many years, quantitative analysis has been seriously complicated by experimental problems (see 2, 8, 10). There has been no preparation as suitable and no drug as specific for Ca channels, as the squid axon and TTX are for Na channels (10). The advent of the isolated adult heart cell and the development of powerful new electrophysiological methods, however, have opened up the possibility to record Ca channel activity at the level of single cells and single channels and to obtain evidence on molecular mechanisms of Ca channel gating and modulation.

Fast and slow gating behaviour of single calcium channels in cardiac cells. Relation to activation and inactivation of calcium-channel current.

The Ca-channel gating behaviour during steady and stepwise depolarization was examined in recordings of single Ca-channel activity from cell-attached membrane patches of single ventricular cells isolated enzymatically from hearts of adult guinea pigs. The single-channel recordings were performed by means of the improved patch-clamp technique (Hamill et al. 1981) with 90 mM Ba in the pipettes. Upon step depolarization, two types of current records were regularly observed in the ensembles: (1) traces with Ca-channel activity (in the form of closely-spaced brief pulses of inward current with a unitary amplitude) of various length, and (2) blank sweeps without any detectable single-channel opening. The records with Ca-channel activity show a distinct tendency for openings to occur towards the beginning of the clamp pulse, followed by long periods of silence. The blank sweeps seem to reflect a condition or conditions where the Ca channel is unavailable for opening. The corresponding ensemble mean current I(t) displayed a rapid rising phase to its peak followed by a slow decay. During steady depolarization, kinetic analysis of the distributions of all open and shut lifetimes revealed a monoexponential probability density distribution function of all open times. By contrast, more than two exponential terms were required for an accurate description of the frequency distribution of all shut lifetimes. Corresponding to the two well-separated fast closed time components, individual Ca-channel openings were grouped into bursts of openings. The bursting behaviour reflected fast gating transitions and was related to the fluctuations of the Ca channel between two short-lived closed states and one open state. This fast gating was terminated by the entrance of the Ca channel into at least one long-lived closed state, exit from which was slow in comparison to the rapid cycling. As consequence, bursts of openings were further grouped together in clusters of bursts, the cluster behaviour being related to slow gating transitions in the kinetics of the Ca channel. The biphasic frequency distribution of the first latencies (resulting from the transit through the two short-lived shut states, before the open state is entered) superimposed on the first time derivative of the rising phase of the ensemble mean current, I(t), upon step depolarization. The time constant of the monoexponential distribution function of all cluster lifetimes matched the declining phase of I(t) during maintained depolarization.(ABSTRACT TRUNCATED AT 400 WORDS)

Modulation of calcium current density by intracellular calcium in isolated guinea pig ventricular cardiomyocytes.

This study describes quantitatively the relation between L-type calcium current (ICa) density and resting cytosolic free calcium concentration ([Ca2+]i) in isolated guinea pig ventricular cardiomyocytes. ICa was measured simultaneously with [Ca2+]i at 22 and 35 degrees C using the whole-cell patch-clamp method in combination with the Fura-2 ratio-fluorescence technique. At 22 degrees C, the increase in [Ca2+]i upon replacing external Na+ with tetraethylammonium resulted in a monotonic decrease in ICa density. Half-maximal inhibition of ICa density occurred at a [Ca2+]i value (K0.5) of 60 +/- 17 nM (mean +/- S.D., n = 59). At 35 degrees C, with a holding potential of -80 mV, a similar relation between [Ca2+]i and ICa was observed (K0.5 of 62 +/- 12 nM, n = 34). When the cells were depolarized from -40 mV, however, the initial decrease in ICa density with increasing [Ca2+]i was usually followed by transient stimulation (< or = 2-fold), which was again overcome by inhibition. Stimulation was not observed in the presence of broad spectrum protein kinase inhibitors. These data suggest that Ca2+i-dependent facilitation of cardiac ICa is mediated by a temperature-sensitive enzymatic pathway that ends in voltage-dependent phosphorylation of Ca2+ channels.

Trypsin and forskolin decrease the sensitivity of L-type calcium current to inhibition by cytoplasmic free calcium in guinea pig heart muscle cells.

A key feature of trypsin action on ionic membrane currents including L-type Ca2+ current (ICa) is the removal of inactivation upon intracellular application. Here we report that trypsin also occludes the resting cytoplasmic free Ca2+ ([Ca2+]i)-induced inhibition of peak ICa in isolated guinea pig ventricular cardiomyocytes, using the whole-cell patch clamp in combination with the Fura-2 ratio-fluorescence technique. The effectiveness of trypsin to guard ICa against [Ca2+]i-induced inhibition was compared with that of forskolin, as cAMP-dependent phosphorylation had been suggested to confer protection against [Ca2+]i-induced inactivation. Intracellular dialysis of trypsin (1 mg/ml) augmented ICa by 7.2-fold, significantly larger than the threefold increase induced by forskolin (3 microM). Forskolin application after trypsin dialysis did not further enhance ICa. An increase in [Ca2+]i from resting levels (varied by 0.2, 10, and 40 mM EGTA dialysis) to submicromolar concentrations after replacement of external Na+ (Na(o)+) with tetraethylammonium (TEA+) resulted in monotonic inhibition of control ICa, elicited from a holding potential of -40 mV at 22 degrees C. AFter trypsin dialysis, however, ICa became less sensitive to submicromolar [Ca2+]i; the [Ca2+]i of half-maximal inhibition (K0.5, normally around 60 nM) increased by approximately 20-fold. Forskolin also increased the K0.5 by approximately threefold. These and accompanying kinetic data on ICa decay are compatible with a model in which it is assumed that Ca2+ channels can exist in two modes (a high open probability "willing" and a low open probability "reluctant" mode) that are in equilibrium with one another. An increase in [Ca2+]i places a larger fraction of channels in the reluctant mode. This interconversion is hindered by cAMP-dependent phosphorylation and becomes nearly impossible after tryptic digestion.

Modulation of L-type Ca2+ current by fast and slow Ca2+ buffering in guinea pig ventricular cardiomyocytes.

Free Ca2+ near Ca2+ channel pores is expected to be lower in cardiomyocytes dialyzed with bis-(o-amino-phenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA) than with ethyleneglycol-bis-(beta-aminoethyl)-N,N,N',N'-tetraacetic acid (EGTA) because BAPTA chelates incoming Ca2+ more rapidly. The consequences of intracellular Ca2+ buffering by BAPTA (0.2-60 mM) and by EGTA (0.2-67 mM) on whole-cell L-type Ca2+ current (ICa,L) were investigated in voltage-clamped guinea pig ventricular cardiomyocytes; bulk cytoplasmic free Ca2+ (Cac2+) was monitored using the fluorescent Ca2+ indicator indo-1. ICa,L was augmented by approximately 12-fold when BAPTA in the cell dialysate was increased from 0.2 to 50 mM (half-maximal stimulation at 31 mM), whereas elevating internal EGTA from 0.2 to 67 mM increased ICa,L only by approximately 2-fold. Cac2+ was < 20 nM with internal BAPTA or EGTA > or = 20 mM. While EGTA up to 67 mM had only an insignificant inhibitory effect on the stimulation of ICa,L by 3 microM forskolin, ICa,L in 50 mM BAPTA-dialyzed myocytes was insensitive to forskolin-induced elevation of adenosine 3',5'-cyclic monophosphate (cAMP); conversely, ICa,L in cAMP-loaded cells was unresponsive to BAPTA dialysis. Cell dialysis with BAPTA, but not with EGTA, accelerated the slow component of ICa,L inactivation (tau S) without affecting its fast component (tau F), resembling the effects of cAMP-dependent phosphorylation. BAPTA-stimulated ICa,L was inhibited by acetylcholine and by the cAMP-dependent protein kinase (PKA) blocker H-89. These results suggest that BAPTA-induced lowering of peri-channel Ca2+ stimulates cAMP synthesis and channel phosphorylation by disinhibiting Ca(2+)-sensitive adenylyl cyclase.

Elementary currents through Ca2+ channels in guinea pig myocytes.

Elementary Ca2+ and Ba2+ currents were recorded from cell-attached membrane patches of ventricular myocytes from adult guinea pig hearts using the improved patch-clamp technique (Hamill et al. 1981). High concentrations of Ba2+ or Ca2+ (50 or 90 mM) were used in the pipettes to increase the signal-to-noise ratio. All data were derived from elementary current analyses in patches containing only one channel. 1) In response to voltage steps, channel openings occurred singly or in bursts of closely spaced unitary current pulses separated by wider shut intervals. During depolarizations of small amplitude from the resting potential, channel openings occurred almost randomly, whereas during larger depolarizations the events were grouped preferentially at the beginning. 2) Channel openings became more probable with increased depolarization; simultaneously, unitary current amplitudes declined in an ohmic manner. Elementary current amplitudes were slightly larger, when 50 mM Ba2+ replaced 50 mM Ca2+ in the pipettes (slope conductances 9 and 10 pS, respectively), but more than doubled, when Ba2+ was increased to 90 mM (slope conductance 18 pS). Clear outward currents through Ca2+ channels were not observed under these conditions. 3) Peak amplitudes of reconstructed mean currents doubled when 50 mM Ba2+ replaced 50 mM Ca2+ and were larger still when 90 mM Ba2+ was used in the pipettes. The current-voltage relations of the reconstructed mean currents showed a positive shift along the voltage axis as Ba2+ was increased or substituted equimolarly by Ca2+. correspondingly, the open state probability-voltage relations (activation curves) showed a parallel shift as Ba2+ was increased, which was less pronounced when Ba2+ was replaced equimolarly by Ca2+. 4) Determination of Ca2+ channel inactivation using 90 mM Ba2+ in the pipettes indicated an overlap with channel activation in a limited voltage range, resulting in a steady-state "window" current. Inactivation can occur without divalent cation influx. 5) Formation of an inside-out patch resulted in a fast rundown of elementary Ca2+ channel currents. 6) Channel openings were often grouped in bursts. The lifetimes of the open state, the bursts, and the closed states were estimated for Ba2+ and Ca2+ as permeating ions. At least two exponentials were needed to fit the histogram of the lifetimes of all closed states. The lifetimes of the individual openings and bursts were mono-exponentially distributed. The kinetics of the Ca+ channel depended on the voltage and the permeating ion. During +30 mV depolarizations, no significant effect on the permeating ion on channel gating could be detected.(ABSTRACT TRUNCATED AT 400 WORDS)

Cat ventricular muscle treated with D600: effects on calcium and potassium currents.

In single sucrose-gap experiments on cat ventricular muscle strands stimulated with 300 ms pulses at 0.33 Hz, 2 microM-D600 reduced the Ca-dependent slow inward current (ICa) by 50% within 5 min and more than 90% in 90-120 min. The late outward current was reduced by up to 30%. During the exposure to D600, Ca channels could be unblocked by hyperpolarizing pulses and blocked again by stimulation with depolarizing pulses. Since the degree of unblocking depended on voltage, and the degree of blocking depended on stimulation pattern, ICa amplitude could be rapidly manipulated to probe the dependence of K conductance on ICa. Under control conditions, an increase in stimulation rate from 0.02 to 1 Hz reduced ICa by 15% and increased the late outward current by a smaller amount. During exposure to D600, a similar intervention provoked a 60% reduction in ICa, but a control-like increase in the late outward current. Two other series of experiments failed to disclose a link between ICa and K conductance: when a block of Ca channels was reimposed following their unblocking, the outward currents were independent of ICa amplitude. Unblock-block experiments also provided information on the extent of steady-state ICa at 0 mV. The fraction of Ca channels not undergoing inactivation appears to be very small. During full D600 block, the inward peak of the current wave form is broad and very much delayed in comparison with pre-drug currents or currents on the first pulse following unblocking. A similar wave form was recorded in D600-treated ventricular myocytes from cat but not guinea-pig. The likely explanation is that D600 unmasks a small transient outward current in cat ventricle.