The effects of 14 days of spaceflight on myonuclear number, fiber size, and myosin heavy chain (MHC) expression in isolated rat soleus muscle fiber segments were studied. Single soleus muscle fibers from rats flown on the Spacelab Life Sciences-2 14-day mission were compared with those from age-matched ground-based control rats by using confocal microscopy and gel electrophoresis. Spaceflight resulted in a significant reduction in the number of fibers expressing type I MHC and an increase in the number of fibers expressing type IIx or IIa MHC. Space-flight also resulted in an increase in the percentage of fibers coexpressing more than one MHC and in the reexpression of the neonatal isoform of MHC in some fibers. Fiber cross-sectional area was significantly reduced in pure type I MHC-expressing fibers and in fibers coexpressing type I+II MHC but not in fibers expressing one or more type II MHC in the flight rats. The number of myonuclei per millimeter was significantly reduced in type I MHC-expressing fibers from the flight rats but was not significantly different in type I+II and type II MHC-coexpressing fibers. Fibers expressing neonatal MHC were similar in size to control fibers but had significantly fewer myonuclei per millimeter than flight fibers not expressing neonatal MHC. In type I MHC-expressing fibers, the reduction in fiber cross-sectional area was greater than the reduction in myonuclear number; thus the average cytoplasmic volume per myonucleus was significantly lower in flight than in control fibers. The reduction in both myonuclear number and fiber size of fibers expressing type I MHC after 14 days of spaceflight supports the hypothesis that changes in the number of myonuclei may be a contributing factor to the reduction in fiber size associated with chronic unloading of the musculature.

We tested the hypothesis that rat soleus muscle fiber growth and changes in myosin phenotype during the postnatal, preweaning period would be largely independent of weight bearing. The hindlimbs of one group of pups were unloaded intermittently from postnatal day 4 to day 21: the pups were isolated from the dam for 5 h during unloading and returned for nursing for 1 h. Control pups were either maintained with the dam as normal or put on an alternating feeding schedule as described above. The enlargement of mass (approximately 3 times), increase in myonuclear number (approximately 1.6 times) and myonuclear domain (approximately 2.6 times), and transformation toward a slow fiber phenotype (from 56 to 70% fibers expressing type I myosin heavy chain) observed in controls were inhibited by hindlimb unloading. These properties were normalized to control levels or higher within 1 mo of reambulation beginning immediately after the unloading period. Therefore, chronic unloading essentially stopped the ontogenetic developmental processes of 1) net increase in DNA available for transcription, 2) increase in amount of cytoplasm sustained by that DNA pool, and 3) normal transition of myosin isoforms that occur in some fibers from birth to weaning. It is concluded that normal ontogenetic development of a postural muscle is highly dependent on the gravitational environment even during the early postnatal period, when full weight-bearing activity is not routine.

Osteoporosis due to mineral loss is a major health problem resulting from long-term spaceflight. The development of a suitable countermeasure is essential because an advanced decrease in bone density could be irreversible. Therefore the current study was performed to test our hypothesis that the loading of bones by electrical stimulation-induced muscle contraction may prevent the mineral loss caused by gravitational unloading and bone growth will be maintained. During 10 d of hindlimb suspension, electrical stimulation at 1, 50, or 100 Hz was administered through the left sciatic nerve at the gluteal region of rats with approximately 300 g body weight. The dry weight, mineral content, and mineral density in hindlimb bones were analyzed. The dry weight and mineral content of femur and tibia-fibula in hindlimb-suspended rats tended to be less than in the age-matched cage controls. However, these detrimental effects were prevented by stimulation at 50 and 100 Hz. A positive effect of stimulation was seen even in the nonstimulated limb, although greater effect was induced in the stimulated limb. It is suggested that loading by stimulation-induced muscle contraction at higher frequencies is beneficial for the maintenance of bone growth or the prevention of mineral loss, or both, during hindlimb suspension in rats.

The effects of chronic stretching or shortening of the soleus muscle of adult rats during hindlimb suspension on muscle mass and contractile properties were studied. Rats suspended with the ankle joint immobilized in either a dorsiflexed (Susp-DF. soleus stretched), a plantarflexed position (Susp-PF, soleus shortened), or without immobilization (Susp-Free. soleus shortened) were compared with cage control rats. Suspension-related muscle atrophy was prevented in Susp-DF. The relative muscle weight in Susp-PF was also less than in cage control and Susp-DF. Both isometric maximum twitch tension (Pt) and maximum tetanic tension (Po) in the Susp-Free and Susp-PF were less than control. Both Pt and Po in Susp-DF were normal. The twitch time-to-peak tension and one-half relaxation time tended to be reduced by chronic shortening of the muscle. The rate of tension development during a twitch (dp/dt), expressed as g/s, of Susp-Free group was decreased, but that expressed as g/s/g Pt was greater than controls. That in Susp-DF was subnormal. The fatigue resistance in Susp-Free was normal but was reduced in Susp-DF and Susp-PF. These data suggest that the decreases in the rat soleus mass and maximum tension production and the shift toward a fast-twitch type following hindlimb suspension are prevented by chronic stretching of muscle, although detrimental effect was induced for the fatigue resistance.

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