Acetylcholine receptors in cultured human muscle cells

Franklin, Gillian I.; Yasin, Rose; Hughes, B.R.; Thompson, Edward J.

doi:10.1016/0022-510x(80)90085-4

Cited by 11 publications

(9 citation statements)

References 16 publications

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“…For assessment of r binding, samples were harvested as above into PBS containing 1% BSA. Binding of ~2sI-a-bungamtoxin (Amersham International) was determined in suspension, and otherwise according to Franklin et al (14), and the specific binding found by adding an excess of unlabeled ct-bungarotoxin (Sigma Chemical Co.). tions were spotted onto nitrocellulose (0.45 I~m, Schleicher & Schuell, Inc., Keene, NH) using a Bin Dot apparatus (Bio-Rad Laboratories).…”

Section: Creatine Kinase Assay and A-bungarotoxin Bindingmentioning

confidence: 99%

Skeletal muscle neural cell adhesion molecule (N-CAM): changes in protein and mRNA species during myogenesis of muscle cell lines.

Moore¹,

Thompson²,

Kirkness³

et al. 1987

The Journal of Cell Biology

124

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Abstract. Qualitative and quantitative changes in neural cell adhesion molecule (N-CAM) protein and mRNA forms were measured during myogenesis in G8-1 and C2 cell lines. Indirect immunofluorescence assay showed that N-CAM was constitutively expressed by myoblasts in culture and that myotubes appeared to be stained more strongly. These changes were quantified using a dot blot assay. N-CAM levels increased almost 4-fold in G8-1 cells and 15-fold in C2 cells during myogenesis. The kinetics of accumulation of N-CAM were not coordinate with other muscle markers such as creatine kinase or acetylcholine receptor levels, since N-CAM accumulated significantly ahead of these markers. Immunoblotting showed that myogenesis was not associated with changes in the extent of sialylation of N-CAM. However, distinct changes in desialo forms were observed after neuraminidase treatment. Myogenesis was accompanied by increases in 125-and 155-kD desialo forms with minor changes in 120-and 145-kD forms. Biosynthetic labeling studies showed that myoblasts specifically expressed a transmembrane isoform of 145 kD that was phosphorylated and was down-regulated in myotubes. Pulse-chase analysis of myotubes showed that the 120-kD isoform and an isoform of 145 kD that co-migrated with, but was distinct from, the 145 kD transmembrane isoform of myoblasts were precursors of the 125-and 155-kD isoforms, respectively, that accumulated in myotubes. The 125-and 155-kD isoforms in myotubes are linked to the cell membrane via phosphatidylinositol linkage and can be released by phospholipase C. Indirect immunofluorescence analysis showed that phosphatidylinositol specific phospholipase C specifically released N-CAM from the myotube membrane generating N-CAM-free myotubes, while myoblasts were unaffected by this treatment. Three N-CAM mRNA species were observed in mouse muscle cell lines. Myoblasts were characterized by their expression of 6.7-and 5.2-kb transcripts while myotubes express 5.2-and 2.9-kb transcripts. Thus, myogenesis is qualitatively associated with a down regulation of the 6.7-kb transcript and an up regulation of the 5.2-and 2.9-kb transcript.

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Section: Creatine Kinase Assay and A-bungarotoxin Bindingmentioning

confidence: 99%

Skeletal muscle neural cell adhesion molecule (N-CAM): changes in protein and mRNA species during myogenesis of muscle cell lines.

Moore¹,

Thompson²,

Kirkness³

et al. 1987

The Journal of Cell Biology

124

View full text Add to dashboard Cite

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“…This process is nerve independent (SYTKOWSKI et al 1973;PRIVES et al 1976;BEKOFF and BETZ 1976;JACOB and LENTZ 1979;FAMBROUGH 1979;FRANKLIN et al 1980;WALSH and RITTER 1981), but muscle activity increases both total AChR and the number of AChR clusters (COHEN and FISCHBACH 1973;SHAINBERG et al 1976). This process is nerve independent (SYTKOWSKI et al 1973;PRIVES et al 1976;BEKOFF and BETZ 1976;JACOB and LENTZ 1979;FAMBROUGH 1979;FRANKLIN et al 1980;WALSH and RITTER 1981), but muscle activity increases both total AChR and the number of AChR clusters (COHEN and FISCHBACH 1973;SHAINBERG et al 1976).…”

Section: A) Acetylcholine Receptors and Acetylcholinesterasementioning

confidence: 99%

Skeletal Muscle

Schmalbruch

1985

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“…If DMD results from some abnormality of the plasma membrane, then this abnormality may also be reflected in changes in the distribution of various cell surface receptors. Franklin et al 41 have used '251labeled a-bungarotoxin (a-bgt) to examine the distribution of acetylcholine receptors in cells from 4 patients with DMD, 2 with Becker dystrophy, and 12 patients with other neuromuscular disorders. The binding of a-bgt increased rapidly at the time of myotube formation, but there was variation between myotubes within the same culture, between different cultures, and between disorders.…”

Section: Distribution Of Acetylcholine Receptorsmentioning

confidence: 99%

“…Nevertheless, binding to myotubes in DMD cultures was generally low, but myotubes in "clusters" (see Cell-Cell Interactions in DMD Muscle Cultures, below) bound a-bgt, whereas the grain density of most of the cells within a cluster was similar to background. Franklin et al 41 reported that the degree of differentiation of myotubes in DMD cultures (measured by creatine kinase activity) was poor, and it is probable that the low levels of a-bgt binding in these cultures reflects this poor differentiation, rather than a defect in the DMD plasma membrane.…”

Section: Distribution Of Acetylcholine Receptorsmentioning

confidence: 99%

Tissue culture studies of muscle disorders: Part 1. Techniques, cell growth, morphology, cell surface

Witkowski¹

1986

Muscle and Nerve

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Tissue culture has been used extensively in studies of human inherited disorders, and its application in the field of the neuromuscular disorders has increased rapidly in recent years. This review, covering the period 1977 to 1984, deals with tissue culture studies of both human and animal muscle disorders, although Duchenne muscular dystrophy (DMD) figures prominently because of the overwhelming interest in that disorder. The review is in two parts. In the first part, I discuss technical innovations in the field, the morphology and growth of cells, and a variety of studies related to the cell surface. Important findings in relation to DMD include reports of abnormal growth rates and reduced lifespan of DMD cells, hypersensitivity to DNA-damaging agents, abnormal cell-to-cell and cell-to-substratum adhesion, and a more "fluid" cell membrane. However, these findings are controversial or have so far been reported only by single laboratories.

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Acetylcholine receptors in cultured human muscle cells

Cited by 11 publications

References 16 publications

Skeletal muscle neural cell adhesion molecule (N-CAM): changes in protein and mRNA species during myogenesis of muscle cell lines.

Skeletal muscle neural cell adhesion molecule (N-CAM): changes in protein and mRNA species during myogenesis of muscle cell lines.

Skeletal Muscle

Tissue culture studies of muscle disorders: Part 1. Techniques, cell growth, morphology, cell surface

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