Expression of small heat shock protein alphaB-crystallin is induced after hepatic stellate cell activation

Using the differential PCR display method to select cDNA fragments that are differentially expressed after hepatic stellate cell (HSC) activation, we have isolated from activated HSCs a cDNA that corresponds to rat alphaB-crystallin. Northern blots confirmed expression of alphaB-crystallin in cultur...

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Bibliographic Details
Published in:American journal of physiology: Gastrointestinal and liver physiology 2000-12, Vol.279 (6), p.G1333-G1342
Main Authors: Lang, A, Schrum, L W, Schoonhoven, R, Tuvia, S, Solís-Herruzo, J A, Tsukamoto, H, Brenner, D A, Rippe, R A
Format: Article
Language:English
Subjects:
Animals
Cells, Cultured
Crystallins - biosynthesis
Crystallins - genetics
Hot Temperature
HSP70 Heat-Shock Proteins - biosynthesis
Liver - metabolism
Liver Cirrhosis, Experimental - metabolism
Male
Molecular Sequence Data
Phosphorylation
Polymerase Chain Reaction
Rats
Rats, Sprague-Dawley
RNA, Messenger - metabolism
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Summary:Using the differential PCR display method to select cDNA fragments that are differentially expressed after hepatic stellate cell (HSC) activation, we have isolated from activated HSCs a cDNA that corresponds to rat alphaB-crystallin. Northern blots confirmed expression of alphaB-crystallin in culture-activated HSCs but not in quiescent HSCs. Western blot analysis and immunocytochemical staining confirmed expression of alphaB-crystallin protein in activated but not quiescent HSCs. alphaB-crystallin is induced as early as 6 h after plating HSCs on plastic and continues to be expressed for 14 days in culture. Expression of alphaB-crystallin was also induced in vivo in activated HSCs from experimental cholestatic liver fibrosis. Confocal microscopy demonstrated a cytoplasmic distribution of alphaB-crystallin in a cytoskeletal pattern. Heat shock treatment resulted in an immediate perinuclear redistribution that in time returned to a normal cytoskeletal distribution. The expression pattern of alphaB-crystallin was similar to that of HSP25, another small heat shock protein, but differed from the classic heat shock protein HSP70. Therefore, alphaB-crystallin represents an early marker for HSC activation.
ISSN:0193-1857