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Am J Physiol Gastrointest Liver Physiol 279: G1333-G1342, 2000;
0193-1857/00 $5.00
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Vol. 279, Issue 6, G1333-G1342, December 2000

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

Alon Lang1, Laura W. Schrum2, Robert Schoonhoven3, Shmuel Tuvia4, Jose A. Solís-Herruzo1, Hidekazu Tsukamoto5, David A. Brenner1,6, and Richard A. Rippe1

Departments of 1 Medicine, 2 Surgery, 3 Environmental Science and Engineering, and 6 Biochemistry and Biophysics, University of North Carolina, Chapel Hill 27599; and 4 Department of Cell Biology, Duke University, Durham, North Carolina 27710; and 5 Departments of Medicine and Pathology, University of Southern California, Los Angeles, California 90033

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 alpha B-crystallin. Northern blots confirmed expression of alpha B-crystallin in culture-activated HSCs but not in quiescent HSCs. Western blot analysis and immunocytochemical staining confirmed expression of alpha B-crystallin protein in activated but not quiescent HSCs. alpha B-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 alpha B-crystallin was also induced in vivo in activated HSCs from experimental cholestatic liver fibrosis. Confocal microscopy demonstrated a cytoplasmic distribution of alpha B-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 alpha B-crystallin was similar to that of HSP25, another small heat shock protein, but differed from the classic heat shock protein HSP70. Therefore, alpha B-crystallin represents an early marker for HSC activation.

differential polymerase chain reaction display; gene expression; liver fibrosis


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