How dose the gastric surface mucus cell line derived from transgenic mouse synthesize and secrete mucins?

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GSM06 is a cell line established from the stomach of transgenic mouse harboring a temperature-sensitive simian virus 40 (SV40) large T-antigen gene. ^3H-Labeled macromolecules produced by the cells incubated with [^3H] glucosamine were characterized to examine whether or not GSM06 cells synthesize mucin (mucus glycoprotein). The GSM06 cells grew until a confluent monolayr formed at 33ﾟC (the permissive temperature for SV40 large T-antigen expression), and the ^3H-labeled macromolecules appeared in both cell extract and medium during culture for at least one week. Unexpectedly, almost all ^3H-labeled macromolecules, which were excluded from a column of Sepharose CL-4B,were identified as hyaluronan by analyzes using Sepharose CL-2B chromatography, cesium trifluoroacetate equilibrium centrigugation, treatment with dithiothreitol, and trypsin and hyaluronidase digestion. At a nonpermissive temperature (39ﾟC), GSM06 cells grew only slightly, but produced much more hyaluronan than at 33ﾟC.The results indicate that GSM06 cells do not produce mucin, but hyaluronan, and that the expression of large T-antigen may influence hyaluronan synthesis in GSM06 cells.