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Am J Physiol Cell Physiol 274: C595-C602, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 3, C595-C602, March 1998

alpha -Cardiac-like myosin heavy chain as an intermediate between MHCIIa and MHCIbeta in transforming rabbit muscle

Heidemarie Peuker, Agnès Conjard, and Dirk Pette

Faculty of Biology, University of Konstanz, D-78457 Constance, Germany

To elucidate the sequence of myosin heavy chain (MHC) transitions in fast-to-slow transforming rabbit muscle, direct reverse transcriptase-polymerase chain reaction was applied for detecting mRNAs specific to five MHC isoforms in single fibers from control and low-frequency-stimulated tibialis anterior muscles. The detection of MHCIIb, MHCIId(x), MHCIalpha , and MHCIbeta mRNAs was based on previously published methods. The RT-PCR assay for MHCIIa mRNA was based on the identification of a cDNA sequence in the 3'-region from which specific primers were derived. Comparisons between rat, rabbit, and human MHCIIa sequences revealed high degrees of sequence identities. MHC mRNA isoform patterns in single fibers from stimulated muscles showed hybrid fibers expressing the following combinations: MHCIId(x) + MHCIIa, MHCIId(x) + MHCIIa + MHCIalpha , MHCIId(x) + MHCIIa + MHCIalpha  + MHCIbeta , MHCIIa + MHCIalpha , MHCIIa + MHCIalpha  + MHCIbeta , and MHCIalpha + MHCIbeta . The combination MHCIIa + MHCIbeta without MHCIalpha was never seen. These coexpression patterns suggest that the fast-to-slow fiber transition results from sequential isoform expressions in the order MHCIId(x) right-arrow MHCIIa right-arrow MHCIalpha right-arrow MHCIbeta . The allocation of MHCIalpha between MHCIIa and MHCIbeta seems to be in line with graded differences in sequence identity of the 3'-regions of these mRNA isoforms.

fiber type transformation; MHCIIa sequence; rabbit skeletal muscle; reverse transcriptase-polymerase chain reaction; single fiber analysis


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