Restoration of junctional tetrads in dysgenic myotubes by dihydropyridine receptor cDNA.
Excitation-contraction coupling was restored in primary cultures of dysgenic myotubes by transfecting the cells with an expression plasmid encoding the rabbit skeletal muscle dihydropyridine receptor. Dishes containing normal, dysgenic, and transfected myotubes were fixed, freeze-fractured, and replicated for electron microscopy. Numerous small domains in the surface membrane of normal myotubes contain ordered arrays of intramembrane particles in groups of four (tetrads). The disposition of tetrads in the arrays is consistent with alternate positioning of tetrads relative to the underlying feet of the sarcoplasmic reticulum. Dysgenic myotubes have no arrays of tetrads. Some myotubes from successfully transfected cultures have arrays of tetrads with spacings equal to those found in normal myotubes. Thus the dihydropyridine receptor appears to be needed for the formation of tetrads and their association with the sarcoplasmic reticulum feet. This result is consistent with the hypothesis that each tetrad is composed of four dihydropyridine receptors.
Takekura
H
H
Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia 19104-6058.
Bennett
L
L
Tanabe
T
T
Beam
K G
KG
Franzini-Armstrong
C
C
D000818:Animals; D000831:Animals, Newborn; D015220:Calcium Channels / Q000502:physiology; D020746:Calcium Channels, L-Type; D002478:Cells, Cultured; D018076:DNA, Complementary; D005614:Freeze Fracturing; D051379:Mice; D008854:Microscopy, Electron; D008961:Models, Structural; D009124:Muscle Proteins / Q000096:biosynthesis / Q000502:physiology*; D009132:Muscles / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D010957:Plasmids; D011817:Rabbits; D014162:Transfection
D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D018076:DNA, Complementary; D009124:Muscle Proteins