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Alcohol Inhibition of Cell Adhesion in BMP‐Treated NG108‐15 Cells
Author(s) -
Wilkemeyer Michael F.,
Pajerski Mandy,
Charness Michael E.
Publication year - 1999
Publication title -
alcoholism: clinical and experimental research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 153
eISSN - 1530-0277
pISSN - 0145-6008
DOI - 10.1111/j.1530-0277.1999.tb04065.x
Subject(s) - cell adhesion , neural cell adhesion molecule , transfection , microbiology and biotechnology , chemistry , cell adhesion molecule , cell , cell culture , adhesion , cell growth , biochemistry , biology , gene , genetics , organic chemistry
Background The L1 cell adhesion molecule is expressed as alternatively spliced neuronal and nonneu‐ronal isoforms. We have reported that in transfected fibroblasts, ethanol variably inhibits cell‐cell adhesion mediated by the nonneuronal isoform of human L1. In contrast, ethanol consistently inhibits morphoge‐netic changes and cell‐cell adhesion in NG108‐15 cells treated with OP‐1 (BMP‐7), a powerful inducer of L1 and N‐CAM gene expression. Methods All studies were performed by using NG108‐15 cells cultured in serum‐free medium. Cell morphology was assessed by a quantitative assay of cell clustering. Cell adhesion was measured by a short‐term re‐aggregation assay, and isoforms of L1 were characterized by RT‐PCR and sequencing. Results We show that ethanol inhibits the morphogenetic effects of BMP‐2, BMP‐4, BMP‐5, and BMP‐6, each of which increases the expression of L1 and N‐CAM. Pretreatment of NG108‐15 cells with 25‐100 mM ethanol did not induce tolerance to ethanol's inhibition of OP‐1 morphogenesis or cell‐cell adhesion. Ethanol or anti‐L1 Fab fragments partially inhibited cell‐cell adhesion in OP‐1‐treated NG108‐15 cells. The combination of ethanol and Fab fragments did not inhibit cell‐cell adhesion more than Fab fragments alone. As in L1‐transfected fibroblasts, a series of n‐alcohols displayed a cutoff between butanol and pentanol for inhibition of cell‐cell adhesion in OP‐1‐treated NG108‐15 cells. RT‐PCR and direct sequencing revealed that the neuronal isoform was the sole or predominant L1 isoform in OP‐1‐treated NG108‐15 cells. Conclusions These data suggest that ethanol inhibits cell‐cell adhesion in OP‐1‐treated NG108‐15 cells by interacting directly or indirectly with the neuronal isoform of L1.