Abstract
Cell adhesion molecules (CAMs) are cell surface glycoproteins that may play a variety of roles in morphogenesis and histogenesis, particularly in defining borders of discrete cell populations. To examine the influence of CAM expression on such cell segregation events in vitro, we have transfected cells with cDNAs coding for two calcium-dependent CAMs of different specificity, the liver CAM (L-CAM) and the structurally related molecule N-cadherin. The cDNAs were introduced separately or together into murine sarcoma S180 cells, which normally do not express these molecules, to produce cell lines denoted S180L, S180cadN, and S180L/cadN, respectively. A number of cell lines of each type were produced that differed in their levels of CAM expression. In adhesion assays, S180L and S180cadN cells aggregated specifically via their respective CAMs, and S180L cells did not appear to adhere to S180cadN cells. Cells expressing high levels of each CAM aggregated more rapidly than cells expressing low levels. Segregation between two cell types occurred when they expressed CAMs of different specificity or different levels of the same CAM. S180L and S180cadN cells both sorted out from untransfected cells, and cells expressing high levels of either L-CAM or N-cadherin segregated from cells expressing low levels of the same CAM; in all cases segregation was inhibited by antibodies specific for the transfected CAM. S180L cells sorted out from S180cadN cells, but this segregation was inhibited only when antibodies to both CAMs were applied together. Doubly transfected S180L/cadN cells also sorted out from S180L cells and from S180cadN cells, and the process was inhibited by antibodies to the unshared CAM (N-cadherin or L-CAM, respectively). Cytochalasin D and nocodazole inhibited sorting-out, consistent with the probable role of microfilaments and microtubules in cell movement and in accord with evidence that the action of these CAMs depends on interactions with cortical cytoplasmic components. Using cDNAs for only two CAMs in these studies, we could distinguish at least eight cell lines by their behavior in sorting-out assays. This suggests that qualitative and quantitative differences in the expression in vivo of a relatively small number of CAMs can lead to a large variety of patterns among cell collectives and their borders during tissue formation.
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