Heterologous expression of BI Ca2+ channels in dysgenic skeletal muscle.
We have examined the ability of BI (class A) Ca2+ channels, cloned from rabbit brain, to mediate excitation-contraction (E-C) coupling in skeletal muscle. Expression plasmids carrying cDNA encoding BI channels were microinjected into the nuclei of dysgenic mouse myotubes grown in primary culture. Ionic currents and intramembrane charge movements produced by the BI channels were recorded using the whole-cell patch-clamp technique. Injected myotubes expressed high densities of ionic BI Ca2+ channel current (average 31 pA/pF) but did not display spontaneous contractions, and only very rarely displayed evoked contractions. The expressed ionic current was pharmacologically distinguished from the endogenous L-type current of dysgenic skeletal muscle (Idys) by its insensitivity to the dihydropyridine antagonist (+)-PN 200-110. Peak BI Ca2+ currents activated with a time constant (tau a) of approximately 2 ms and inactivated with a time constant (tau h) of approximately 260 ms (20-23 degrees C). The time constant of inactivation (tau h) was not increased by substituting Ba2+ for Ca2+ as charge carrier, demonstrating that BI channels expressed in dysgenic myotubes do not undergo Ca(2+)-dependent inactivation. The average maximal Ca2+ conductance (Gmax) produced by the BI channels was quite large (approximately 534 S/F). In contrast, the average maximal charge movement (Qmax) produced in the same myotubes (approximately 2.7 nC/microF) was quite small, being barely larger than Qmax in control dysgenic myotubes (approximately 2.3 nC/microF). Thus, the ratio Gmax/Qmax for the BI channels was considerably higher than previously found for cardiac or skeletal muscle L-type Ca2+ channels expressed in the same system, indicating that neuronal BI Ca2+ channels exhibit a much higher open probability than these L-type Ca2+ channels.
Adams
B A
BA
Department of Physiology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523.
Mori
Y
Y
Kim
M S
MS
Tanabe
T
T
Beam
K G
KG
D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism*; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004558:Electric Stimulation; D015640:Ion Channel Gating; D008564:Membrane Potentials; D051379:Mice; D009119:Muscle Contraction; D018482:Muscle, Skeletal / Q000002:abnormalities / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010957:Plasmids; D011817:Rabbits
D015220:Calcium Channels; D002118:Calcium