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C Cognard

Publications and source records attributed to C Cognard.

At least 91 records · Page 5Linked to original sources

Calcium current-dependent staircase in rat myotubes and myoballs developing in culture.

Calcium current and contraction were simultaneously recorded in whole-cell patch-clamped rat skeletal muscle cells grown in primary culture. Repetitive depolarizations at low frequency, which elicited calcium currents, led to a staircase response, characterized by the progressive increase of both twitch amplitude and activation rate. It was sometimes possible to elicit a staircase response in 2 or 3 day old postfusion myotubes which did not or weakly contract initially. The staircase response was dependent on calcium entry through calcium channels, since it was reversed when calcium current was depressed by means of inorganic calcium blockers or depolarization to large positive potential. The entry of calcium was also necessary to allow the development of a staircase response following caffeine-induced contractures which partly emptied the intracellular stores of calcium. These features are consistent with the idea that calcium currents allow the initial loading of intracellular calcium stores and, in later stages, serve to replenish and maintain them constant.

Animals↗

Appearance and evolution of calcium currents and contraction during the early post-fusional stages of rat skeletal muscle cells developing in primary culture.

Primary cultures from enzymatically dissociated satellite cells of newborn rat skeletal muscles enabled developmental in vitro studies of mechanical and electrical properties during the first steps of myogenesis. The present work focused on the appearance, evolution and roles of two types of calcium currents (ICa,T and ICa,L) and of depolarization-induced contractile activity during the early stages of muscle cell development in primary culture. Prefusional mononucleated cells (myoblasts), young myotubes of 1 day (with less than 10 nuclei) or 2-3 days (more than 9 nuclei) and myoballs from 4-6, 7-9, 10-12 and 13-16 days cultures were patch-clamped (whole-cell configuration), and calcium currents and contraction simultaneously recorded. Sodium but not calcium currents could be recorded at the myoblast stage. In young myotubes (1 day), ICa,L was present with high incidence as compared to ICa,T, which was poorly expressed. Contractile responses appeared at the next stage (2-3 days) while the occurrence of ICa,T progressively increased. This developmental evolution of the calcium currents and contraction expression was accompanied by some changes in their characteristics: the ICa,T/ICa,L amplitudes ratio progressively increased and the time-to-peak of contraction progressively decreased with the age of myoballs. Physiological functions for calcium currents in developing muscle are suggested and discussed: ICa,T, which is transiently expressed, could be involved in the pacemaker-like activity while ICa,L could serve as an early contraction triggering mechanism and/or initially to fill and then to maintain the intracellular calcium stores.

Animals↗

A third type of calcium current in cultured human skeletal muscle cells.

A third type of calcium current could be recorded on a non-negligible number of human skeletal muscle cells (normal and Duchenne dystrophic (DMD)) in primary culture. This transient current exhibited a maximum at 0 mV, a time-to-peak around 30 ms, an inactivation time constant around 70 ms and was insensitive to nifedipine. On these basis, it differentiates from T- and L-type previously described and looks like the neuronal N-type. However, this third type of current was not sensitive to omega-Cgtx, a specific N-type blocker. The occurrence and the possible role of this current are briefly discussed.

Calcium Channel Blockers↗

Progressive predominance of 'skeletal' versus 'cardiac' types of excitation-contraction coupling during in vitro skeletal myogenesis.

Simultaneous recordings, in the presence or absence of cadmium, of current and contraction of skeletal muscle cells in primary culture (myoballs) showed that the relative part of contraction depending on calcium current progressively decreases with the age of cells whereas the cadmium-insensitive component becomes predominant. The coexistence of "cardiac" and "skeletal" excitation-contraction coupling mechanisms in developing muscle and its developmental regulation are consistent with recent molecular data showing the expression, in newborn animals, of an abbreviated isoform of the alpha 1 subunit of the DHP-receptor of muscle cells.

Animals↗

The correlation between the increase in slow outward current and in contraction induced by caffeine, ryanodine, and rapid cooling in voltage-clamped frog muscle fibers.

The effects of caffeine, ryanodine, and rapid cooling were tested on the depolarization-induced contraction and the apamin-insensitive slow outward current (Iso) of voltage-clamped (double mannitol gap) single frog muscle fibers. Subthreshold caffeine concentrations (0.5-2 mM) induced a monotonic increase in contractile and Iso amplitude. Whatever the concentration, the increase in contraction was roughly twice the one in current. Similar results were obtained upon rapid cooling (20-4 degrees C) in the presence of 0.5 mM caffeine. In the absence of external Na+ (choline-substituted) 10(-5) M ryanodine induced a delayed increase (approximately 30 min) in contraction and in current, shortly before the development of a drastic and irreversible contracture. Here again, the increase in contraction was twice that in current. In the presence of 5 mM tetraethylammonium (TEA) and (or) 25 nM charybdotoxin, 2 mM caffeine still induced a strong facilitating effect on contraction but the parallel increase in current was strongly reduced. The linear relationship between the increase in current and contractile amplitude has a slope approximately 0.5 (whatever the drug used to increase contractility); it is approximately 0.1 in the presence of TEA and (or) charybdotoxin. In conclusion, provided the changes in contractile amplitude are caused by parallel changes in depolarization-induced sarcoplasmic reticulum Ca2+ release, about 50% of the apamin-insensitive Iso is controlled by internal Ca2+ release. The main part of this current corresponds to the TEA- and charybdotoxin-sensitive component of Iso.

Animals↗

The blockade of excitation/contraction coupling by nifedipine in patch-clamped rat skeletal muscle cells in culture.

The effects of the dihydropyridine derivative, nifedipine, well known as a blocker of calcium channels, were tested on cultured rat myoballs. Membrane currents and contractions were simultaneously recorded by means of the patch-clamp technique and a photoelectric transducing method. High concentrations of nifedipine (5 microM) inhibited the contractile responses and inward calcium current (ICa) elicited by long depolarizations. In the absence of ICa (1.5 mM cadmium in the bath), nifedipine inhibited both the ICa-independent contractile component and the outward current, supposed to depend on the intracellular calcium released during contraction. At low concentrations (0.5 microM) the blocking effects of nifedipine could be strongly enhanced by shifting the membrane potential towards less negative values (-60 mV) for 50 s prior to the test pulse. A blocking effect of nifedipine, at a usually ineffective concentration (0.1 microM), could also be observed when long-lasting (3 min) prepulses to 0 mV were applied from a reference membrane potential of -60 mV. This effect could be relieved by long-lasting cell hyperpolarizations (-90 mV). The blocking effects of nifedipine unrelated to ICa could be interpreted as an action on a molecule (voltage sensor) in the T-tubule membrane involved in the excitation/contraction coupling process and as a preferential binding of the dihydropyridine derivative on the inactivated form of this molecule, favored by the weak negative potentials or long-lasting depolarizations. The results provide data in favor of the existence of strong similarities between the calcium channels and voltage sensors since their operation was inhibited in a voltage-dependent manner by nifedipine.

Animals↗

The depressing effect of tetracaine and ryanodine on the slow outward current correlated with that of contraction in voltage-clamped frog muscle fibres.

The effects of tetracaine (10-50 microM) and ryanodine (0.1-10 microM) were tested on the slow outward K+ current (Iso) and the mechanical tension of isolated frog muscle fibres in a voltage-clamp device (double mannitol-gap) connected to a mechanoelectric transducer. In the concentration range tested, both drugs induced a simultaneous inhibition of tension and current. In all cases the effect on tension was twice that on current. The tetracaine-induced current and tension blocks were fully reversible and dose-dependent. In contrast the ryanodine effects on current and tension were not reversible and did not exhibit a dose dependence except for the delay before the onset of the response, which was shortened when the concentration was raised. Linear regression analysis of the time-dependent and dose-dependent effects of both drugs indicated a strong correlation between the decreases in tension and current. It is concluded that the slow outward current is partly under the control of the Ca2+ release from sarcoplasmic reticulum during contraction.

Alkaloids↗

Phenytoin preferentially inhibits L-type calcium currents in whole-cell patch-clamped cardiac and skeletal muscle cells.

The effect of the anticonvulsant diphenylhydantoin (phenytoin) was tested on the inward calcium currents of whole-cell patch-clamped cells from rat and human muscles and from frog atrium. A concentration of 10 microM phenytoin was required to obtain a threshold inhibitory effect and, even with high concentrations (100 microM), the inhibition was not complete. In skeletal muscle (rat and human cells in culture), phenytoin (30 microM) exerted a more potent effect on the high-threshold calcium current (ICa,L inhibition: 53 +/- 6% mean +/- SDn-1) rather than on the low-threshold one (ICa,T inhibition: 16 +/- 10%). Similar results were obtained on dissociated frog atrial cells. These data are to be contrasted with those previously reported on neuronal cells, where specific inhibition of ICa,T was reported. Thus, the action of phenytoin appears to be different in muscle and nerve so that phenytoin does not appear to be a specific inhibitor of ICa,T.

Animals↗

Calcium currents in normal and dystrophic human skeletal muscle cells in culture.

Human muscle cells obtained from biopsy specimens were grown in a primary culture system and electrophysiologically studied. Whole cell patch-clamp recordings revealed the presence of two types of calcium currents: (i) a low-threshold (-60 mV) one (ICa, T) with fast activation and inactivation kinetics (time-to-peak: 39 ms at -30 mV); and (ii) a high-threshold (-10 mV) one (ICa,L) with slower kinetics (time-to-peak: 550 ms at 20 mV). These two types of calcium currents could be also distinguished by their pharmacological characteristics since ICa,L was sensitive to the antagonist and agonist dihydropyridine derivatives contrary to ICa,T which was completely resistant to these compounds. These functional calcium channels existed both in normal and Duchenne dystrophic (DMD) human skeletal muscle cells in culture. We discuss a possible role of these two types of calcium channels in the myoplasmic calcium accumulation observed in the Duchenne muscular dystrophy.

Calcium↗

The depressant effects of some amiloride analogues on the slow outward K+ current and contraction of voltage-clamped frog muscle fibres.

The effects of 4 derivatives of amiloride, which are known to block the Na+/H+ antiporter, were studied on the slow outward current (Iso) and on tension development of voltage-clamped single muscle fibres of the frog Rana ridibunda. Each compound tested induced a strong depressant effect on tension and Iso in a voltage-, time- and dose-dependent (10(-8) to 10(-5) M) manner at physiological pH. The effects exhibited a strong pH dependence (the greater the pH in the range between 6.6 and 8.0, the greater the depressant action). This seems to exclude the involvement of the Na+/H+ antiport and alterations of surface membrane charges as mediators. The current block was not observed after the release of calcium from the sarcoplasmic reticulum was blocked with 75 microM tetracaine. It is concluded that the primary effect of the 4 compounds used was to inhibit the internal release of Ca2+ from the sarcoplasmic reticulum, and that this inhibition blocked the Ca2+-dependent slow outward current.

Amiloride↗

The slow inward calcium current is responsible for a part of the contraction of patch-clamped rat myoballs.

The slow inward calcium current and the contractile response were simultaneously recorded in voltage clamped (whole cell patch clamp recording) rat myoballs in primary culture. The shape of the contraction(T)/potential(V) relationship and the application of the inorganic calcium channel blocker cadmium (1.5 mM), which suppresses a part of the contractile activity, demonstrate the existence of two components of contraction. One of them is related to the slow calcium current.

Animals↗

Pharmacological and biochemical characterization of rat hippocampal 5-hydroxytryptamine1A receptors solubilized by 3-[3-(cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS).

Rat hippocampal 5-hydroxytryptamine1A (5-HT1A) binding sites were solubilized with a yield of 34% using 3-[3-(cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS, 10 mM) as detergent. Kinetic analyses of [3H]8-hydroxy-2-(di-n-propylamino)tetralin ([3H]8-OH-DPAT) binding indicated that the 5-HT1A sites exhibit the same properties in the soluble form as in the membrane-bound form. Furthermore, a positive correlation (r = 0.988) was found between the respective pIC50 values of a series of agonists and antagonists to inhibit [3H]8-OH-DPAT binding to either soluble or membrane-bound 5-HT1A sites. Gel filtration through Sephacryl S-400 as well as chromatography on wheat germ agglutinin (WGA)-agarose did not affect the modulation by guanine nucleotides (5'-guanylylimidodiphosphate) of [3H]8-OH-DPAT binding which suggests that the 5-HT1A binding subunit is a glycoprotein tightly attached to a G protein even in its soluble form. The [3H]8-OH-DPAT binding material eluted from Sephacryl S-400 had an apparent molecular mass of 155 kilodaltons, as expected from a heterodimer with one binding subunit (approximately 60 kilodaltons) and one G protein (approximately 80 kilodaltons). Marked enrichment in 5-HT1A binding sites relative to other soluble proteins was found in the peak fractions eluted from Sephacryl S-400 (by sixfold) and WGA-agarose (by 26-fold) columns, suggesting that these chromatographic steps might be of interest for the purification of central 5-HT1A receptors.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Antamanide antagonizes the phalloidin-induced negative inotropic effect and blocks voltage dependently the fast outward K+ current in voltage-clamped frog muscle fibres.

The effects of antamanide (10(-14)-10(-5) M) and N-acetyl-secophalloidin (10(-7)-5 X 10(-3) M) a neutral non-toxic derivative of phalloidin, were tested on voltage-clamped single frog muscle fibres. Antamanide protected muscle fibres against the negative inotropic effect of phalloidin but blocked the fast potassium permeability in the same concentration range and the same voltage-dependent manner as did phalloidin. N-Acetyl-secophalloidin exhibited a strongly attenuated blocking effect on K+ permeability in a 1,000-fold higher concentration range than phalloidin. Neither antamanide nor N-acetyl-secophalloidin affected the contractile properties. These results suggest the existence in the frog muscle membrane of a receptor with two sites for phalloidin and antamanide which acts on potassium conductance.

Animals↗

Stimulation frequency and external ionic composition control the repriming of caffeine-induced contractures in frog skeletal muscle.

The effect of stimulation rate and of external ionic composition on the repriming period of contractures induced by 6 mM caffeine was tested on isolated skeletal muscle fibres of the frog (Rana ridibunda). The repriming period, which was 11.2 +/- 0.1 min (mean +/- SEM, n = 9) on quiescent fibres, was shortened in fibres stimulated at a frequency ranging from 3 to 12 min-1 (optimal rate, 8 min-1; full repriming 5.7 +/- 0.2 min; n = 10). A 10-fold increase in the extracellular calcium concentration shortened the repriming period on both stimulated and quiescent fibres, whereas decreasing external calcium (1/10) delayed it. In a Na+-free solution (Li+ substituted) the repriming period of stimulated fibres was markedly delayed (14 min), whereas quiescent fibres never recover more than 10% of their ability to develop subsequent caffeine contractures. In contrast, with a 35% Na solution, the repriming period was greatly shortened (stimulated, 5.4 +/- 0.2 min, n = 7; quiescent, 6.2 +/- 0.5 min, n = 8). It is concluded that repriming depends on three mechanisms that seem to refill a calcium store and trigger recovery: the slow inward calcium current, a Na+-Ca2+ exchange, and perhaps a passive Ca2 influx.

Animals↗

[Two components of calcium-dependent potassium permeability coexist in the membrane of frog skeletal muscle].

Two components of Ca-dependent K permeabilities were demonstrated in the membrane of voltage-clamped muscles fibres by raising the internal calcium concentration with 3 mM caffeine or 0.1 microM Bay K 8644. One, which is TEA-sensitive, is enhanced by an increase of intracellular calcium and the other, which is apamin-sensitive, is inhibited by high levels of internal calcium concentration.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗