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Volume 271, Number 39, Issue of September 27, 1996 pp. 24089-24095
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Differential Expression of B-Crystallin and Hsp27 in Skeletal Muscle during Continuous Contractile Activity
RELATIONSHIP TO MYOGENIC REGULATORY FACTORS

(Received for publication, April 17, 1996, and in revised form, June 26, 1996)

From the Department of Internal Medicine, Molecular Cardiology Research Laboratories, University of Texas Southwestern Medical Center, Dallas, Texas 75235-8573

B-crystallin (BC) is a major structural protein (22 kDa) of the ocular lens as well as a bona fide heat shock protein in non-lens tissue. The BC gene is abundantly expressed in tissues with high oxidative capacity, including the heart and type I skeletal muscle fibers, and is regulated by the MyoD family of basic helix-loop-helix transcription factors during myogenesis. To test the hypothesis that BC expression may be directly regulated by the demand for oxidative metabolism, we examined the expression of BC and the ancestral-related Hsp27 in rabbit tibialis anterior muscle subjected to continuous low frequency motor nerve stimulation (3 V, 10 Hz). BC mRNA and protein increased within the 1st day of continuous contractile activity (5- and 2.5-fold, respectively) and achieved maximum levels (>20-and 4-fold, respectively) after 21 d of stimulation. Hsp27 mRNA and protein levels also increased with stimulation, but with a less specific and dramatic induction pattern. In agreement with the Northern analysis, in situ hybridization performed on cross sections from tibialis anterior muscle revealed progressively increasing BC transcript signal, localized in a ringlet pattern, from 1 through 21 days of stimulation. Serial sections subjected to myosin immunohistochemistry revealed that BC expression was confined to slow-twitch type I and a subpopulation of fast twitch type II fibers after 1 day but present in nearly all fibers after 21 days of stimulation. Transcript levels of all four myogenic regulatory factors (MyoD, myogenin, myf-5, and MRF4) also increased with stimulation in a pattern temporally similar with BC, suggesting that expression of BC in response to stimulation may, in part, be regulated through myogenic regulatory factor(s) interaction with the canonical E-box element located within the BC promotor. These data demonstrate that expression of the small heat shock protein, BC, is rapidly induced independent of the ancestrally related Hsp27 in a fiber type specific pattern in skeletal muscle subjected to the oxidative stress imposed by continuous contractile activity.

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