MRF4, Myf-5, and myogenin mRNAs in the adaptive responses of mature rat muscle.

We studied the possible role of specific muscle regulatory factors (MRF) in the adaptive response to changes in contractile activity in mature skeletal muscle. The tibialis anterior muscle of anesthetized female rats was subjected to low-frequency stimulation, static stretch, or a combination of both. Message levels of MRF were observed after 2 h of activity, and the subsequent 20-h recovery period by slot blot and in situ hybridizations for MRF4, Myf-5, and myogenin. A combination of stimulation and stretch for 2 h increased MRF4 (11.6 +/- 5.3-fold) and Myf-5 (6.6 +/- 1.4-fold). In situ hybridization showed abundance in some regions of the muscle with positive staining near peripheral nuclei of both large and small fibers. Message levels remained high for 30 min and declined to near control levels by 20 h of recovery. Myogenin mRNA levels were unaffected by any manipulations. Neither stretch alone nor 10 Hz of electrical stimulation alone induced a significant increase in MRF. We conclude that myonuclei, and possibly activated myoblasts, increase expression of Myf-5 and MRF4 after a combination of both stimulation and stretch for 2 h.

IIx and slow myosin expression follow mitochondrial increases in transforming muscle fibers.

Metabolic profile and contractile isoform expression commonly define classic fiber types in skeletal muscle. Little is known about how metabolic requirements determine expression of fast IIx and slow myosin isoforms in muscles undergoing fiber type conversion. Tibialis anterior muscles from female New Zealand White rabbits were stimulated continuously at 10 Hz for 4-21 days. Quantitative fiber analysis was made for oxidative potential by histochemistry and for fast IIx and slow myosin mRNA content by in situ hybridization. In control muscle we found 3 +/- 0.27% fibers coexpress both fast IIx and slow myosin mRNA and so were not assignable to a classic fiber type. After stimulation, increase in fiber oxidative potential was detectable by 4 days and preceded IIx mRNA increases on a fiber-by-fiber basis. Slow myosin transcripts were detected by 7 days in fibers with higher oxidative levels. Coexpression of IIx and slow transcripts peaked at 22 +/- 2.5% of fibers by 7 days. IIx then declined, leaving slow myosin expressed in 62 +/- 0.45% of fibers by 3 wk. We conclude that during fiber type transformation individual fibers can transcribe two myosin mRNAs synchronously. Metabolic demand precedes and may be linked to IIx and slow myosin isoform expression.

Repair of injured skeletal muscle: a molecular approach.

We review cellular and molecular processes involved in injury and repair of skeletal muscle with regard to the amount and location of damage produced. Discussion is based on advances made by use of newer techniques, including immunochemistry, in situ hybridization, molecular biology, ultrastructural analysis, and cell culture. Damage and repair processes after eccentric work, stretch, overload, chronic stimulation, cold injury, and other models are discussed for cellular and molecular components. Hypertrophy or hyperplasia can occur under certain conditions. After injury, satellite cells are activated by growth factors. These cells can also be activated during fiber-type transformation, probably to provide necessary DNA content rather than to supply cells of a new lineage. Emphasis is given to myosin mRNA studied by in situ hybridization to localize subcellular distribution. Increases in mRNA concentration are found near nuclei in damaged regions and at the subcellular sites being repaired in the middle of skeletal muscle fibers or near the myotendon junction. The activation of genes for muscle regulatory factors during development is compared with their activation in regeneration and response to injury.