The membrane skeleton of acetylcholine receptor domains in rat myotubes contains antiparallel homodimers of beta-spectrin in filaments quantitatively resembling those of erythrocytes.
I used immunogold labeling and quick-freeze, deep-etch, rotary replication to characterize the membrane skeleton at regions with high concentrations of acetylcholine receptor domains in receptor clusters of cultured rat muscle cells. This membrane skeleton consists of a network of filaments closely applied to the cytoplasmic membrane surface. The filaments are specifically decorated by immunogold labeling with a monoclonal antibody, VIIF7, that recognizes an isoform of beta-spectrin colocalizing with acetylcholine receptors. The filaments are 32 +/- 11 nm in length and three to four filaments (average 3.1-3.3) join at each intersection to form the network. These parameters are nearly identical to those reported previously for the membrane skeleton of erythrocytes. Depending on the amount of platinum coating, filament diameters range from 9 to 11 nm in diameter, and are 1.4 nm larger on average than spectrin filaments of erythrocytes replicated at the same time. Filaments are decorated with gold particles close to one end, consistent with the location of the epitope recognized by the monoclonal antibody. Computer modeling shows that all filament intersections in the membrane skeletal network are equally capable of being labeled by the monoclonal antibody. This pattern of labeling is consistent with a network containing antiparallel homodimers of beta-spectrin.
Department of Anatomy, University of Maryland at Baltimore 21201, USA.
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Pumplin
D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000648:ultrastructure*; D004912:Erythrocytes / Q000648:ultrastructure*; D005613:Freeze Etching; D046911:Macromolecular Substances; D016253:Microscopy, Immunoelectron; D009132:Muscles / Q000737:chemistry / Q000648:ultrastructure*; D017434:Protein Structure, Tertiary; D051381:Rats; D011950:Receptors, Cholinergic / Q000737:chemistry*; D013049:Spectrin / Q000737:chemistry*
D046911:Macromolecular Substances; D011950:Receptors, Cholinergic; D013049:Spectrin