Abstract
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 αB-crystallin. Northern blots confirmed expression of αB-crystallin in culture-activated HSCs but not in quiescent HSCs. Western blot analysis and immunocytochemical staining confirmed expression of αB-crystallin protein in activated but not quiescent HSCs. αB-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 αB-crystallin was also induced in vivo in activated HSCs from experimental cholestatic liver fibrosis. Confocal microscopy demonstrated a cytoplasmic distribution of αB-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 αB-crystallin was similar to that of HSP25, another small heat shock protein, but differed from the classic heat shock protein HSP70. Therefore, αB-crystallin represents an early marker for HSC activation.
Original language | English |
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Pages (from-to) | G1333-G1342 |
Journal | American Journal of Physiology - Gastrointestinal and Liver Physiology |
Volume | 279 |
Issue number | 6 42-6 |
DOIs | |
State | Published - 2000 |
Externally published | Yes |
Keywords
- Differential polymerase chain reaction display
- Gene expression
- Liver fibrosis