Membrane-bound acetylcholinesterase: an early differentiation marker for skeletal myoblasts.
Cell-bound acetylcholinesterase (AChE) was found to be an early differentiation marker on embryonic chick skeletal myoblasts in mixed primary cell cultures. AChE biosynthesis was detected and characterized by (a) a sensitive microtiter assay, (b) use of selective inhibitors, and (c) with mono- and polyclonal antibodies. Both secreted and cell-bound AChE appeared on the first day in culture, at a time when no muscle cell fusion was observed. Characterization of this enzyme revealed that true AChE was bound and secreted by myoblasts. BW284c51, which permeates cell membranes poorly, inhibited all the cell-associated AChE activity on myoblasts, suggesting that the activity measured was on the outer cell surface. On the other hand, fibroblasts appeared to have no or very little bound enzyme and the low level of secreted enzyme activity had the characteristics of pseudo-, or butyrylcholinesterase. Polyclonal anti-Torpedo californica electroplax AChE antibody and several monoclonal antibodies were found to bind specifically to chick myoblasts. Since the cells had not been made permeable before antibody binding, a membrane-bound form of the enzyme was most likely being detected. The cell-bound true AChE was present in identifiable quantities from the first day of culture. Membrane-bound AChE can thus serve as an early differentiation marker for embryonic chick myoblasts in mixed primary cultures.
Elson
H F
HF
Department of Biological Chemistry, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
Gentry
M K
MK
Doctor
B P
BP
D000110:Acetylcholinesterase / Q000032:analysis* / Q000276:immunology; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002091:Butyrylcholinesterase / Q000032:analysis; D002454:Cell Differentiation; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D002642:Chick Embryo; D009132:Muscles / Q000196:embryology* / Q000201:enzymology
D000906:Antibodies; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase