Document Detail

Identification of osmosensitive and ammonia-regulated genes in rat astrocytes by Northern blotting and differential display reverse transcriptase-polymerase chain reaction.
MedLine Citation:
PMID:  11592597     Owner:  NLM     Status:  MEDLINE    
BACKGROUND/AIMS: The hepatic encephalopathy (HE) is in part the result of astrocyte swelling with alterations of glial function. Detoxification of ammonia may be one mechanism by which astrocyte swelling is triggered in HE. METHODS: The differential display polymerase chain reaction (DDRT-PCR) and Northern blot analysis were used for study the functional consequence of cell volume changes and ammonia on gene expression in primary rat astrocytes. RESULTS: Differentially expressed cDNA products were identified with about 92% homology to genes coding for mouse proline rich protein expressed in brain (PRTB), rat clusterin, elongin, and human Kelch motif containing protein. As shown by Northern blot analysis, PRTB and clusterin mRNA levels were upregulated by 19-64% after 4-8 h by both ammonia and hypoosmolarity. Elongin mRNA expression increased by 97% in response to ammonia but slightly by hypoosmolarity. Further, hypoosmotic exposure for 1-24 hours but not ammonia led to an increase of 80% in KMCP mRNA levels. CONCLUSIONS: The identification of these genes offers the opportunity to identify unrecognized molecular mechanisms of HE. The finding that several genes are induced by both, hypoosmolarity and ammonia, supports the view that astrocyte swelling is a major, but not the only pathogenetic event in HE.
U Warskulat; S Kreuels; H W Müller; D Häussinger
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of hepatology     Volume:  35     ISSN:  0168-8278     ISO Abbreviation:  J. Hepatol.     Publication Date:  2001 Sep 
Date Detail:
Created Date:  2001-10-10     Completed Date:  2002-02-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8503886     Medline TA:  J Hepatol     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  358-66     Citation Subset:  IM    
Clinic for Gastroenterology, Hepatology and Infectiology, Heinrich-Heine-University, Düsseldorf, Germany.
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MeSH Terms
Ammonia / pharmacology*
Astrocytes / metabolism*
Base Sequence
Blotting, Northern
Cell Size
Cells, Cultured
Gene Expression Regulation*
Glycoproteins / genetics
Molecular Chaperones / genetics
Molecular Sequence Data
Osmolar Concentration
Rats, Wistar
Reverse Transcriptase Polymerase Chain Reaction
Transcription Factors / genetics
Reg. No./Substance:
0/Clusterin; 0/Glycoproteins; 0/Molecular Chaperones; 0/Transcription Factors; 0/elongin; 7664-41-7/Ammonia

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