Mouse 3T3 fibroblasts were permanently transfected with cDNAs encoding isoforms of the neural cell adhesion molecule (N-CAM) present in human skeletal muscle and brain. Parental and transfected cells were then used in a range of adhesion assays. In the absence of external shear forces, transfection with cDNAs encoding either transmembrane or glycosylphosphatidylinositol (GPI)-linked N-CAM species significantly increased the intercellular adhesiveness of 3T3 cells in suspension. Transfection of a cDNA encoding a secreted N-CAM isoform was without effect on adhesion. Cells transfected with cDNAs containing or lacking the muscle-specific domain 1 sequence, a four-exon group spliced into the muscle but not the brain GPI-linked N-CAM species, were equally adhesive in the assays used. We also demonstrate that N-CAM-mediated intercellular adhesiveness is inhibited by 0.2 mg/ml heparin; but, at higher concentrations, reduced adhesion of parental cells was also seen. Coaggregation of fluorescently labeled and unlabeled cell populations was performed and measured by comparing their distribution within aggregates with distributions that assume nonspecific (random) aggregation. These studies demonstrate that random aggregation occurs between transfected cells expressing the transmembrane and GPI-linked N-CAM species and between parental cells and those expressing the secreted N-CAM isoform. Other combinations of these populations tested exhibited partial adhesive specificity, indicating homophilic binding between surface-bound N-CAM. Thus, the approach exploited here allows for a full analysis of the requirements, characteristics, and specificities of the adhesive behavior of individual N-CAM isoforms.
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1 December 1989
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December 01 1989
Intercellular adhesion mediated by human muscle neural cell adhesion molecule: effects of alternative exon use.
J A Pizzey,
J A Pizzey
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
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L H Rowett,
L H Rowett
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
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C H Barton,
C H Barton
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
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G Dickson,
G Dickson
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
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F S Walsh
F S Walsh
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
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J A Pizzey
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
L H Rowett
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
C H Barton
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
G Dickson
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
F S Walsh
Department of Neurochemistry, Institute of Neurology, Queen Square, London, United Kingdom.
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1989) 109 (6): 3465–3476.
Citation
J A Pizzey, L H Rowett, C H Barton, G Dickson, F S Walsh; Intercellular adhesion mediated by human muscle neural cell adhesion molecule: effects of alternative exon use.. J Cell Biol 1 December 1989; 109 (6): 3465–3476. doi: https://doi.org/10.1083/jcb.109.6.3465
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