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.
|Journal||American Journal of Physiology - Gastrointestinal and Liver Physiology|
|Issue number||6 42-6|
|State||Published - 2000|
- Differential polymerase chain reaction display
- Gene expression
- Liver fibrosis