High-resolution en-face visualization of the cardiomyocyte plasma membrane reveals distinctive distributions of spectrin and dystrophin.
The actin-binding proteins, spectrin and dystrophin, are key components of the plasma membrane-associated cytoskeleton of the cardiac muscle cell. From confocal immunofluorescence studies, the distribution of spectrin appears to overlap with that of dystrophin, but the precise functional differentiation, molecular distributions and spatial relationship of these two cytoskeletal systems remain unclear. Freeze-fracture replica immuno-electron microscopy, in parallel with immunofluorescence/confocal microscopy, were applied to examine at high resolution the spatial relationships between the spectrin and dystrophin membrane-associated cytoskeleton systems in cardiac muscle. Application of freeze-fracture replica cytochemistry, with single and double immunogold labeling, permitted simultaneous examination of the organization of spectrin and dystrophin in en-face views of the plasma membrane at high resolution. In contrast to the close spatial relationship previously demonstrated for dystrophin and beta-dystroglycan, no association between the gold label marking dystrophin and that marking spectrin was observed. Our freeze-fracture cytochemical results suggest that the two membrane skeletal networks formed by dystrophin and spectrin in cardiac muscle are independently organized, implying that whatever overlap of function (e.g., in structural support to the plasma membrane) may exist between them, the two systems may each have additional distinctive roles.
Stevenson
Shirley A
SA
Faculty of Medicine, Imperial College London, National Heart and Lung Institute, Guy Scadding Building, UK.
Cullen
Michael J
MJ
Rothery
Stephen
S
Coppen
Steven R
SR
Severs
Nicholas J
NJ
D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D003599:Cytoskeleton / Q000737:chemistry; D016189:Dystrophin / Q000032:analysis*; D005614:Freeze Fracturing; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D032383:Myocytes, Cardiac / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013049:Spectrin / Q000032:analysis*
D016189:Dystrophin; D013049:Spectrin