Glycosaminoglycans and proteoglycans induce gap junction expression and restore transcription of tissue-specific mRNAs in primary liver cultures

Abstract

Normal rat hepatocytes maintained on tissue culture plastic and in serum-supplemented medium lose their gap junctions within 12 hr and expression of their tissue-specific functions within 24 to 72 hr. The gap junctions are lost via internalization and degradation, and the differentiated functions due to loss of synthesis and to rapid degradation of tissue-specific mRNAs. Near normal levels of tissue-specific mRNAs can be achieved by stabilization of the mRNAs but not by transcription (for most genes), if the cells are cultured in a serum-free, hormonally defined medium and on substrata of tissue culture plastic, fibronectin or laminin, or on various purified collagens. The hormonally defined medium also extends the life-span of the gap junctions to about 24 hr. Certain glycosaminoglycans, proteoglycans and anionic polysaccharides have proven to be potent inducers of gap junction expression and function, to increase abundance of tissue-specific mRNAs, and to lower abundance of common gene mRNAs, a level of gap junctions and a pattern of gene expression similar to that in vivo. Addition to the hormonally defined medium of 10 micrograms per ml of hyaluronates, dermatan sulfates, bovine lung heparan sulfate, chondroitin 4-sulfate or chondroitin 6-sulfate resulted in a weak response in induction of gap junctions (5 to 15% of the cells became dye and electrically coupled) and in gene expression. An intermediate response in gap junction expression (30 to 50% coupled cells) and in gene expression was observed with 50 to 100 micrograms per ml of heparins or hyaluronates.(ABSTRACT TRUNCATED AT 250 WORDS)