Effect of β-adrenergic stimulation on the relationship between membrane potential, intracellular [Ca2+] and sarcoplasmic reticulum Ca2+ uptake in rainbow trout atrial myocytes

Anna Llach, Jingbo Huang, Franklin Sederat, Lluis Tort, Glen Tibbits, Leif Hove-Madsen

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    Long depolarizations cause a steady tonic contraction and induce sarcoplasmic reticulum (SR) Ca2+-uptake in trout atrial myocytes. Simultaneous measurements of cytosolic [Ca2+] ([Ca2+]i) and whole membrane current showed an elevated [Ca2+]i throughout the depolarization. Rapid caffeine (Caf) applications at -80 mV before and after a long depolarization were used to determine SR Ca2+ loading and its dependency on membrane potential and [Ca2+]i during depolarization. Following a 10 s depolarization, the maximal SR Ca2+ load was 597 μmol l-1 and loading was half-maximal at -12 mV. The β-adrenergic agonist isoproterenol (ISO) did not affect the maximal SR Ca2+ loading but shifted the potential for half-maximal loading by -26 mV. Following a 3 s depolarization, the maximal SR Ca2+ uptake rate (V̇max) was 418 μmol l-1 s-1 in control conditions. ISO did not affect V̇max, but significantly lowered the average free Ca2+ transient during the depolarization and shifted the K0.5 for the relationship between SR Ca2+ uptake and [Ca2+]i from 1.27 in control to 0.8 μmol l-1 with ISO. Following repetitive 200 ms depolarizations, ISO increased the L-type Ca2+ current (ICa) amplitude by 91±29% and the peak Ca2+ transient by 41±10%, and decreased the half life of the Ca2+ transient from 151±12 to 111±6 ms. Using the relationship between [Ca2+]i and SR Ca2+ uptake to calculate the total SR Ca2+ uptake during a Ca2+ transient elicited by a 200 ms depolarization, a significant increase in the SR Ca 2+ uptake from 37±6 μmol l-1 in control to 68±4 μmol l-1 with ISO was seen. When normalized to the total Ca2+ transport the contribution of the SR was not significantly different in the absence (35±6%) or presence of ISO (41±4%). Exposure of cells to ISO and low extracellular [Ca2+] increased ICa by 67±40% (N=5) but significantly reduced SR Ca 2+ uptake at membrane potentials above -30 mV. Together, these results suggest that (i) ISO has a stimulatory effect on the SR Ca2+ pump that may contribute to the faster decay of the Ca2+ transient, and (ii) the relative contribution of the SR to the Ca2+ removal during relaxation is not altered by ISO in trout atrial myocytes.
    Original languageEnglish
    Pages (from-to)1369-1377
    JournalJournal of Experimental Biology
    Publication statusPublished - 1 Mar 2004


    • Caffeine
    • Excitation-contraction coupling
    • Membrane current
    • Na-Ca exchange
    • Teleost heart
    • Trout

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