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Acetylcholine receptor channels are present in undifferentiated satellite cells but not in embryonic myoblasts in culture
Authors:G Cossu  F Eusebi  F Grassi  E Wanke
Affiliation:1. Duke University Medical Center, Box 104790, Durham 27710, NC, USA;2. Toxicology Program, North Carolina State University, Raleigh, NC, USA;3. Center for Human Heath and the Environment, North Carolina State University, Raleigh, NC, USA;1. Molecular Population Genetics & Breeding Group, Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, 117604, Singapore;2. Shanghai Fisheries Institute, 265 Jiamusi Road, Shanghai 200433, China;3. Department of Fisheries, Rongchang Campus, Southwestern University, 160 Xueyuan Road, Rongchang, Chongqing 402460, China;4. Department of Biological Sciences, National University of Singapore, 14 Science Drive, 117543, Singapore
Abstract:The expression and the physiological properties of acetylcholine receptors (AChRs) of mononucleated myogenic cells, isolated from either embryonic or adult muscle of the mouse, have been investigated using the gigaohm seal patch-clamp technique in combination with immunocytochemistry (with an anti-myosin antibody) and alpha-bungarotoxin binding techniques. Undifferentiated (myosin-negative) embryonic myoblasts, grown either in mass culture or under clonal conditions, were found to be unresponsive to ACh and did not bind alpha-bungarotoxin. On the contrary, undifferentiated satellite cells (from adult muscle) exhibited channels activated by ACh and alpha-bungarotoxin binding sites similar to those observed in differentiated (myosin-positive) embryonic myoblasts and myotubes. Two classes of ACh-activated channels with different opening frequencies were identified. The major class of channels had a conductance of about 42 pS and mean open time of 3.1-8.2 msec. The minor class of channels had smaller conductance (about 17 pS) and similar open time. During differentiation, the conductance of the two channels did not change significantly, while channel lifetime became shorter in myotubes derived from satellite cells but not in myotubes derived from embryonic myoblasts. The relative proportion of small over large channels was significantly larger in embryonic than in adult myogenic cells.
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