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  • 1
    ISSN: 1432-1424
    Keywords: calcium current ; skeletal muscle ; sulfhydryl ; invertebrate muscle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Voltage-clamp experiments using the three-microelectrode voltage clamp technique were performed on ventroabdominal flexor muscles of the crustacean Atya lanipes. Potassium and chloride currents were found to underlie the normal, passive response of the muscle. Blocking potassium currents with tetraethylammonium and replacing chloride ions with methanesulfonate did not unmask an inward current. By treating the muscle with the sulfhydryl-alkylating agent 4-cyclopentene-1,3-dione an inward current was detected. The current induced by the agent is carried by Ca2+, since it is abolished in Ca2+-free solutions. The induced Ca+current is detected at about -40 mV and reaches a mean maximum value of -78 μA/cm2 at ca. -10 mV. At this potential the time to peak is close to 15 msec.The induced Ca2+ current inactivated with 1-sec prepulses which did not elicit detectable Ca2+ current; the fitted h xcurve had a midpoint of-38 mV and a steepness of 5.0 mV. Measurements of isometric tension were performed in small bundles of fibers, and the effects of the sulfhydryl-alkylating agents 4-cyclopentene-1,3-dione andN-ethylmaleimide were investigated. Tetanic tension was enhanced in a strictly Ca2+-dependent manner by 4-cyclopentene1,3-dione. The amplitude of K+ contractures increased after treatment with N-ethylmaleimide. It is concluded that Ca2+ channels are made functional by the sulfhydryl-specific reagents and that the increase in tension is probably mediated by an increase in Ca2+ influxthrough the chemically induced Ca2+ channels.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 173 (2000), S. 9-17 
    ISSN: 1432-1424
    Keywords: Key words: Crustacean muscle — L-type calcium channels — Excitation-contraction coupling — Calcium-induced calcium release
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. The superficial (tonic) abdominal flexor muscles of Atya lanipes do not generate Ca2+ action potentials when depolarized and have no detectable inward Ca2+ current. These fibers, however, are strictly dependent on Ca2+ influx for contraction, suggesting that they depend on Ca2+-induced Ca2+ release for contractile activation. The nature of the communication between Ca2+ channels in the sarcolemmal/tubular membrane and Ca2+ release channels in the sarcoplasmic reticulum in this crustacean muscle was investigated. The effects of dihydropyridines on tension generation and the passive electrical response were examined in current-clamped fibers: Bay K 8644 enhanced tension about 100% but did not alter the passive electrical response; nifedipine inhibited tension by about 70%. Sr2+ and Ba2+ action potentials could be elicited in Ca2+-free solutions. The spikes generated by these divalent cations were abolished by nifedipine. As the Sr2+ or Ba2+ concentrations were increased, the amplitudes of the action potentials and their maximum rate of rise, V max , increased and tended towards saturation. Three-microelectrode voltage-clamp experiments showed that even at high (138 mm) extracellular Ca2+ concentration the channels were silent, i.e., no inward Ca2+ current was detected. In Ca2+-free solutions, inward currents carried by 138 mm Sr2+ or Ba2+ were observed. The currents activated at voltages above −40 mV and peaked at about 0 mV. This voltage-activation profile and the sensitivity of the channels to dihydropyridines indicate that they resemble L-type Ca2+ channels. Peak inward current density values were low, ca.−33 μA/cm2 for Sr2+ and −14 μA/cm2 for Ba2+, suggesting that Ca2+ channels are present at a very low density. It is concluded that Ca2+-induced Ca2+ release in this crustacean muscle operates with an unusually high gain: Ca2+ influx through the silent Ca2+ channels is too low to generate a macroscopic inward current, but increases sufficiently the local concentration of Ca2+ in the immediate vicinity of the sarcoplasmic reticulum Ca2+ release channels to trigger the highly amplified release of Ca2+ required for tension generation.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 409 (1987), S. 555-560 
    ISSN: 1432-2013
    Keywords: Acetylcholine receptor ; Sulfhydryl reagents ; Neuromuscular junction ; Miniature endplate current
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The acethylcholine receptor was chemically modified using bisulfite to add a sulfonate group to a disulfide bond on the α subunit, and diamide, an oxidizing agent, to form an interchain disulfide bond between β subunits of adjacent receptors. In previous work, both reagents increased mepc decay times but produced no change in mean channel open time or conductance as measured by spectral analysis of endplate current fluctuations (Steinacker and Zuazaga 1981). In the current work, we show that, while both chemical modifications increase the decay time of the miniature endplate current, only sulfonation increases the time to peak. Sulfonation also produced an effect on voltage jump current relaxation time, which parallels the increase in miniature endplate current decay time, and an increase in the ratio of the current relaxation amplitudes. Diamide had no effect on voltage jump current relaxation amplitudes or time constants. These data are analyzed in an attempt to correlate changes in specific rate constants to changes in the macroscopic current measurements.
    Type of Medium: Electronic Resource
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