Recovery of mRNA Expression of Tryptophan 2,3-Dioxygenase and Serine Dehydratase in Long-Term Cultures of Primary Rat Hepatocytes

Expression of tryptophan 2,3-dioxygenase (TO) and serine dehydratase (SDH) has not previously been maintained or re-induced in long-term cultured hepatocytes. In the present study, we succeeded in inducing expression of TO and SDH mRNAs in adult rat hepatocytes cultured in serum-free L-15 medium sup...

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Published in:Journal of biochemistry (Tokyo) 1996-09, Vol.120 (3), p.511-517
Main Authors: Mizuguchi, Toru, Mitaka, Toshihiro, Hirata, Koichi, Nakamura, Toshikazu, Mochizuki, Yohichi
Format: Article
Language:English
Subjects:
3-dioxygenase
Animals
Blotting, Northern
Cells, Cultured
Dexamethasone - pharmacology
dimethyl sulfoxide
Dimethyl Sulfoxide - pharmacology
Glucagon - pharmacology
Kinetics
L-Serine Dehydratase - biosynthesis
Liver - drug effects
Liver - enzymology
Male
primary rat hepatocytes
Rats
Rats, Sprague-Dawley
re-expression
RNA, Messenger - biosynthesis
serine dehydratase
Serum Albumin - biosynthesis
Time Factors
Transcription, Genetic - drug effects
tryptophan 2
Tryptophan Oxygenase - biosynthesis
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Summary:Expression of tryptophan 2,3-dioxygenase (TO) and serine dehydratase (SDH) has not previously been maintained or re-induced in long-term cultured hepatocytes. In the present study, we succeeded in inducing expression of TO and SDH mRNAs in adult rat hepatocytes cultured in serum-free L-15 medium supplemented with epidermal growth factor and 2% dimethyl sulfoxide (DMSO). After the start of culture, the expression of TO mRNA rapidly disappeared and at 96 h it was less than 10% of that at isolation. However, after the addition of 2% DMSO from 96 h, the transcript level gradually increased and reached about 40% of that of the isolated cells at day 14. In addition, the expression of TO mRNA was enhanced in cells treated with both 10-5 M dexamethasone and 10-7 M glucagon. In contrast, the expression of SDH mRNA decreased very rapidly and we could not detect it after 24 h of culture. Furthermore, 2% DMSO failed to induce it. However, when both 10-5 M dexameth asone and 10-7 M glucagon were added to the culture medium at day 9, we observed dramatic induction of SDH mRNA 24 h later. Primary hepatocytes cultured by this method could express and maintain highly differentiated hepatic functions for a long time. Thus, this in vitro system is suitable for the investigation of hepatic functions.
ISSN:0021-924X
DOI:10.1093/oxfordjournals.jbchem.a021443