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Biomedical subjects

D Zacharová

Publications and source records attributed to D Zacharová.

At least 19 recordsLinked to original sources
PubMed1993

Modulation by adrenaline of electrophysiological membrane parameters and contractility in intact and internally perfused single muscle fibres of the crayfish.

The effect of catecholamines on basic membrane characteristics (including labeled ionic fluxes) and contractile parameters was followed in current clamp and voltage clamp conditions in intact muscle fibres and internally perfused muscle fibre segments respectively of the crayfish Astacus fluviatilis; i.e. in muscle fibres which spike and activate tension on calcium principle. Both adrenaline and noradrenaline (6.10(-6) mol/l) facilitated twitch tension induced by graded membrane responses or strontium all-or-none spikes. No effect of isoprenaline was observed. Adrenaline (6.10(-6)-6.10(-5) mol/l) produced an inotropic effect, which appeared with a latency of 2 min and reached its maximum in 5 min. The rates of activation and relaxation of contraction were increased, whilst the latency and the threshold depolarization were decreased. The changes persisted (several tens of min) after washout of adrenaline, depending on concentration and duration of adrenaline application. The resting potential and the strontium spike (Ca2+ replaced with Sr2+) were not influenced and the graded responses were facilitated by adrenaline (from 36.4 +/- 1 mV to 40.0 +/- 2 mV; RP = 77.2 +/- 0.5 mV). Extracellular Ca2+ ions are required for the inotropic effect of adrenaline to occur. The decrease of electrical and contractile responses in nominal calcium-free solutions or after a blockade of Ca2+ influx by Ni2+ ions (1 mmol/l) was relieved by adrenaline. The persistence of inotropic effect of adrenaline was absent, when the extracellular concentration of Ca2+ ions, [Ca2+]0 was decreased from 13.5 to 3.4 mmol/l or the Ni2+ ions were added. The influx of 89Sr2+ ions was decreased in the presence of Ni2+ ions from 24.2 +/- 4.7 pmol.cm-2.s-1 to 11.0 +/- 2.8 pmol.cm-2.s-1, but restored to 20.4 +/- 5.8 pmol.cm-2.s-1 in the presence of adrenaline (6 mumol/l). Adrenaline itself decreased the influx of 89Sr2+ ions, and prolonged the time constant of efflux both in resting and stimulated fibres. The effect of adrenaline is dependent on mobilization of Ca2+ ions from the sarcoplasmic reticulum. First, the inotropic effect of adrenaline was absent in the presence of procaine (blockator of the Ca release channel of the SR), in spite of the increase of the active membrane response (all-or-none procaine action potential); second, adrenaline accelerated the uptake of Ca ions by SR as evidenced by shortening of the restitution processes after caffeine contractures by adrenaline. Membrane calcium currents are increased by adrenaline as a rule; mainly at lower depolarizations (-50 to -20 mV).(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1993

Characterization of tension decline in different types of fatigue-resistant skeletal muscle fibres of the frog. Low extracellular calcium effects.

Twitch and tetanic tension have been measured in single skeletal muscle fibres (m.ileofibularis, Rana esculenta and Rana temporaria). On the basis of resistance to fatigue produced by repetitive tetanic stable frequency stimulation with various numbers of stimulation trains, twitch fibers were subdivided in three groups (resembling those as described by Westerblad and Lännergren 1986 in Xenopus), that is fatigue-resistant (FR), moderately fatigued (MF) and easily fatiguing (EF). It was found further that the fibres differ in tetanic tension decline resistance i.e. fatiguability relating to some basic contractile parameters including the amplitude, the rates parameters of twitch and tetanus tension as well as the tetanus/twitch tension ratio. The main differences observed concern: 1) The inability to maintain the maximum tetanic tension plateau (IMT) during single tetanus. IMT was 18 times higher in EF fibres and 4 times higher in MF fibres, respectively, in comparison with FR fibres. IMT is the first parameter to change significantly during repetitive tetanic stimulation. 2) The different fibre types show pronounced differences in twitch contraction and tetanus tension during repetitive tetanic stimulation. There is a conspicuous facilitation of twitch tension during and after cessation of repetitive stimulation in FR fibres; the MF and EF fibres show, on the contrary, a depression of twitches. 3) Recovery to original (prefatigue) values is rapid in FR fibres, but slow, however, in EF fibres. 4) Removal of extracellular Ca2+ intensified the inability to maintain the maximum tetanic tension (IMT) and the tetanic tension decline, especially in fibres with an initial high fatigue-resistance. We assume that the results might be explained by a different refractoriness of transmission between the T-tubules and the sarcoplasmic reticulum in examined fibres and/or by a different dependency of the T-SR transmission on the extracellular calcium ions. A possible cause of the failure may be an intensification of the inactivation process.

Animals↗
PubMed1993

The effect of Pb2+ ions on calcium currents and contractility in single muscle fibres of the crayfish.

Muscle fibres of crustaceans represent an useful model for studying the mode of action of substances influencing calcium channels as the membrane generates the active responses on a pure calcium principle and the excitation-contraction link is dependent on external calcium. We followed the effect of Pb2+ ions (1-300 mumol/l) on contractile responses in single muscle fibres of the crayfish evoked by massive or intracellular electrical stimulation, by potassium depolarization and caffeine application, as well as on action potential in single intact muscle fibres and on calcium currents in voltage clamp conditions (vaseline gap) in internally perfused muscle fibre segments. All types of contractile responses, single twitches, tetanus, potassium and caffeine contractures were blocked. The strontium action potential was blocked very effectively by Pb2+ ions. The total calcium currents which can be split by means of Hodgkin-Huxley equations into two components (fast and slow respectively) differing in the rate of activation and inactivation were suppressed after addition of Pb(NO3)2 (50-300 mumol/l). The effect of Pb2+ was concentration and time dependent. At lower concentrations (100 mumol/l) the blocking effect was more pronounced on the fast inactivating Ca current component. The Pb2+ ions prolonged the time constant of inactivation tau h of the slow channel, while leaving that of the fast channel intact.

Action Potentials↗
PubMed1990

Effects of ruthenium red on excitation and contraction in muscle fibres with Ca2+ electrogenesis.

The effect of ruthenium red (RR) on the electrical and contractile responses, membrane Ca currents, staining patterns of the external and internal membrane system were tested in intact and mechanically skinned muscle fibres of the crayfish Astacus fluviatilis. The following results were obtained: 1. Depression of the contractile responses following membrane depolarization (twitch, tetanus, potassium contractures). 2. Caffeine contractures were unaffected in intact (100 mumol/l - 1 mmol/l RR) and blocked in skinned fibres (30 mumol/l RR). 3. Mechanical threshold and mechanical latency were increased and/or prolonged. 4. The rate of depolarization of the action potentials (AP) was decreased and decremental spread of AP was recorded. 5. Both fast and slowly inactivating Ca ionic currents were decreased and the time constants of activation (tau(m] and inactivation (tau(h] were prolonged after RR (100 mumol/l) pretreatment. 6. The penetration of RR into the T-system was inversely related to its binding to the sarcolemma. The depression of depolarization-induced contractions was most pronounced in fibres with unstained sarcolemma and stained T-tubules. In intact fibres, neither terminal cisternae nor other elements of SR were stained. On the contrary, all internal membrane structures were stained in skinned fibres. There was a gradient of staining intensity from surface toward the interior.

Animals↗
PubMed1990

[Excitation-contraction coupling in isolated muscle fibers with calcium electrogenesis preserved in a culture medium].

The aim of the present work was to check changes in functional characteristics of isolated muscle cells, operating on the calcium electrogenesis principle, while kept in culture media for several days. Skeletal muscle cells of the crayfish Astacus fluviatilis were used to study potassium/caffeine contractures and single/tetanic contractions; simultaneous electrical and mechanical responses were recorded by the microelectrode technique, and kinetics of calcium ionic currents was studied under vaseline-gap voltage clamp. In cultured fibers, active membrane responses and calcium current kinetics remained unchanged, or slightly increased, whereas contractile responses were substantially reduced. A gradual excitation-contraction decoupling was observed. The fiber maintained the ability to respond to direct activation (by caffeine) of the contractile apparatus. Subthreshold caffeine concentrations (0.2-0.5 mmol/l) and adrenaline (6.0(-6), 6.10(-5) mol/l) enhanced the inhibited (due to the culturing) single contractile responses.

Action Potentials↗
PubMed1989

The roles of haemocytes during degeneration and regeneration of crayfish muscle fibres.

Crayfish haemolymph contains three types of haemocytes with cytoplasmic granules: coagulocytes, granulocytes and amoebocytes. Muscle degeneration was induced by either a gross mechanical injury or a mild puncture injury of m. extensor carpopoditi. Granulocytes and amoebocytes were involved in the phagocytosis of disintegrating muscle fibres. Within three weeks after the gross injury the first myotubes were found. The formation of regenerated fibres started before the degenerating material was removed completely. Mild injury resulted in the formation of contraction clots, localized at the ends of a fibre and connected to a persistent external lamina in the form of an empty sheath. The external lamina sheaths were invaded by amoebocytes. They arranged themselves into a superficial layer similar to an epithelium, formed gap junctions and zonulae adherentes, and showed an increase in the number of cytoplasmic microtubules. These transformed haemocytes retained their ability to engulf material of the disintegrating fibre. In about three weeks the number of microtubules in the transformed haemocytes decreased, and newly formed contractile filaments appeared. Satellite cells are present along the normal crayfish muscle fibres. Following their activation in degenerated material, they might conceivably induce the transformation of haemocytes into myogenic cells.

Animals↗
PubMed1989

Calcium channels in skeletal muscle fibres of the frog.

The vaseline gap voltage clamp technique was used to study slow and fast calcium currents in phasic and tonic muscle fibres of the frogs Rana temporaria (R.t.) and Xenopus laevis (X.l.). At physiological Ca2+ and Na+ concentrations three inward currents could be recorded in R.t. twitch muscle fibres: a fast Ca current, a slow Ca current, and a non-specific (INS) current carried by Na ions. The last current appeared as a sole component at a low threshold membrane potential (MP -55 mV) and was present until the slow Ca current appeared. In tonic R.t. muscle and twitch X. laevis fibres only the fast and the slow Ca currents could be recorded; however the inactivation of the slow Ca current was very prolonged in the presence of Na ions. A modulatory effect of Na ions on Ca2+ currents was observed. In phasic fibres the fast Ca currents inactivated either very slowly or rapidly.

Animals↗
PubMed1988

Intracellular site of Sr2+ and Ba2+ accumulation in frog twitch muscle fibres as determined by electron probe X-ray microanalysis.

Strontium and barium can substitute for calcium at different levels of the excitation-contraction-relaxation cycle. The problem of sequestration of these ions in cellular microcompartments may be settled only by direct evidence obtained with analytical methods. Isolated frog twitch muscle fibres were perfused with increasing concentrations of potassium in Ca-free solution supplemented with Sr2+ (10 mmol/l) or Ba2+ (5 mmol/l). After equilibration in a Ca-free Ringer with Sr2+ or Ba2+ for 30 to 60 min the fibres were frozen in liquid propane (at 80 K) to immobilise ions. Ultrathin (150 nm) cryosections were cut at 170 K, freeze-dried, carbon-coated and analysed in an electron microscope equipped with an X-ray spectrometer. The ultrastructure of the superficial layer of the fibres was satisfactorily preserved. The terminal cisternae (t.c.) of the sarcoplasmic reticulum (SR) were dark and contained various amounts of Sr or Ba in addition to Ca. In Sr loaded fibres the longitudinal SR occasionally showed electron dense content with significant amounts of Ca; no Sr was present. The results suggest that t.c. is the common sequestering compartment for Ca, Sr and Ba. Essentially the same distribution pattern of Sr was found following precipitation of Sr with a solution containing digitonin and Koxalate.

Animals↗
PubMed1987

Histochemical study of calcium on T-tubule membranes and in the sarcoplasmic reticulum, in frog twitch muscle fibres at rest and during activity.

The trigger calcium hypothesis of signal transmission between T-tubules and terminal cisternae (TC) of the sarcoplasmic reticulum (SR) in twitch muscle fibres implies the presence of calcium along T-tubule membranes at rest and its release upon excitation. To test this hypothesis, calcium was immobilised using a fixing and precipitating solution of glutaraldehyde in phosphate buffer at pH 8.0 and the calcium was substituted for by lead. Simultaneous tension recordings revealed the occurrence of contractions or a burst of twitches upon perfusion with the fixative. Procaine or tetrodotoxin (TTX) was used to inhibit this activity. In fibres without fixative-induced activity, precipitates were observed along T-tubules and in adjoining parts of TC. In activated fibres, tubular and TC precipitates were absent. These results are consistent with the trigger calcium hypothesis. In fibres activated by depolarisation, calcium returned to TC after passing successively through different parts of the SR.

Animals↗