Cell surface acetylcholinesterase molecules on multinucleated myotubes are clustered over the nucleus of origin.
Multinucleated skeletal muscle fibers are compartmentalized with respect to the expression and organization of several intracellular and cell surface proteins including acetylcholinesterase (AChE). Mosaic muscle fibers formed from homozygous myoblasts expressing two allelic variants of AChE preferentially translate and assemble the polypeptides in the vicinity of the nucleus encoding the mRNA (Rotundo, R. L. 1990. J. Cell Biol. 110:715-719). To determine whether the locally synthesized AChE molecules are targeted to specific regions of the myotube surface, primary quail myoblasts were mixed with mononucleated cells of the mouse muscle C2/C12 cell line and allowed to fuse, forming heterospecific mosaic myotubes. Cell surface enzyme was localized by immunofluorescence using an avian AChE-specific monoclonal antibody. HOECHST 33342 was used to distinguish between quail and mouse nuclei in myotubes. Over 80% of the quail nuclei exhibited clusters of cell surface AChE in mosaic quail-mouse myotubes, whereas only 4% of the mouse nuclei had adjacent quail AChE-positive regions of membrane, all of which were located next to a quail nucleus. In contrast, membrane proteins such as Na+/K+ ATPase, which are not restricted to specific regions of the myotube surface, are free to diffuse over the entire length of the fiber. These studies indicate that the AChE molecules expressed in multinucleated muscle fibers are preferentially transported and localized to regions of surface membrane overlying the nucleus of origin. This targeting could play an important role in establishing and maintaining specialized cell surface domains such as the neuromuscular and myotendinous junctions.
Rossi
S G
SG
Department of Cell Biology and Anatomy, University of Miami School of Medicine, Florida 33101.
Rotundo
R L
RL
D000110:Acetylcholinesterase / Q000096:biosynthesis / Q000187:drug effects / Q000302:isolation & purification*; D000818:Animals; D001692:Biological Transport; D002451:Cell Compartmentation; D002459:Cell Fusion; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D002467:Cell Nucleus / Q000648:ultrastructure; D016764:Cell Polarity; D002478:Cells, Cultured; D017364:Collagenases / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D008565:Membrane Proteins / Q000187:drug effects / Q000302:isolation & purification; D051379:Mice; D009132:Muscles / Q000166:cytology / Q000201:enzymology* / Q000648:ultrastructure; D011487:Protein Conformation; D011784:Quail
D008565:Membrane Proteins; D000110:Acetylcholinesterase; D017364:Collagenases