Adaptation of Mouse Skeletal Muscle to Long-Term Microgravity in the MDS Mission

The effect of microgravity on skeletal muscles has so far been examined in rat and mice only after short-term (5–20 day) spaceflights. The mice drawer system (MDS) program, sponsored by Italian Space Agency, for the first time aimed to investigate the consequences of long-term (91 days) exposure to microgravity in mice within the International Space Station. Muscle atrophy was present indistinctly in all fiber types of the slow-twitch soleus muscle, but was only slightly greater than that observed after 20 days of spaceflight. Myosin heavy chain analysis indicated a concomitant slow-to-fast transition of soleus. In addition, spaceflight induced translocation of sarcolemmal nitric oxide synthase-1 (NOS1) into the cytosol in soleus but not in the fast-twitch extensor digitorum longus (EDL) muscle. Most of the sarcolemmal ion channel subunits were up-regulated, more in soleus than EDL, whereas Ca2+-activated K+ channels were down-regulated, consistent with the phenotype transition. Gene expression of the atrophy-related ubiquitin-ligases was up-regulated in both spaceflown soleus and EDL muscles, whereas autophagy genes were in the control range. Muscle-specific IGF-1 and interleukin-6 were down-regulated in soleus but up-regulated in EDL. Also, various stress-related genes were up-regulated in spaceflown EDL, not in soleus. Altogether, these results suggest that EDL muscle may resist to microgravity-induced atrophy by activating compensatory and protective pathways. Our study shows the extended sensitivity of antigravity soleus muscle after prolonged exposition to microgravity, suggests possible mechanisms accounting for the resistance of EDL, and individuates some molecular targets for the development of countermeasures

The role of sphingolipids in the control of skeletal muscle function: a review.

In this review, potential roles for the endogenous sphingolipid, sphingosine, and its derivatives are described for muscle cells. Sphingosine modulates the function of important calcium channels in muscle, including the ryanodine receptor (RyR) calcium release channel of the sarcoplasmic reticulum (SR). Sphingosine blocks calcium release through the SR ryanodine receptor and reduces the activity of single skeletal muscle RyR channels reconstituted into planar lipid bilayers. Sphingosine-blocked calcium release is coincident with the inhibitory effects of sphingosine on [3H]ryanodine binding to the RyR. The sphingomyelin signal transduction pathway has also been identified in both skeletal and cardiac muscle. A neutral form of sphingomyelinase (nSMase) enzyme has been localized to the junctional transverse tubule membrane. The high turnover of the SMase is responsible for the production of ceramide and sphingosine. HPLC analyses indicate that significant resting levels of sphingosine are present in muscle tissue. A model of excitation-contraction coupling is presented suggesting a potential role for this endogenous sphingolipid in normal muscle function. Putative roles for sphingolipid mediators in skeletal muscle dysfunction are also discussed. We hypothesize that sphingosine plays important roles in malignant hyperthermia and during the development of muscle fatigue

Type 1, 2A, 2B myosin haevy chain electrophoretic analysis of rat muscle fibers

Mammalian skeletal muscles are mixture of three type of fibers: type 1, type 2A, and type 2B fibers. Immunological studies and proteolytic analysis of myosin heavy chains from the three type of fibers have demonstrated the presence of distinct myosin isoforms. By using typed single muscle fibers and improving an electrophoretic method we are able to resolve three distinct polypeptides which are demonstrate to correspond to type 1, 2A and 2B myosin heavy chain isoforms by using specific monoclonal antibodies. The analysis of single muscle fibers shows that different myosin heavy chain isoforms are frequently coexpressed in the same muscle fiber

Calcium sensitivity and myofibrillar protein isoforms of rat skinned skeletal muscle fibres

We investigated the calcium sensitivity for tension generation of different fibre types and the possible correlation between calcium sensitivity and the presence of distinct regulatory protein and myosin light chain (MLC) isoforms in rat skinned skeletal muscle fibres. Fibre types 1, 2A and 2B were identified by electrophoretic analysis of myosin heavy chain (MHC) isoforms. Fibres showing more than one MHC isoform were discarded. Type 1 fibres from the soleus showed a higher pCa (-log10 [Ca], where [ ] denotes concentration) threshold and a lower slope of pCa/tension curve than type 2 extensor digitorum longus (EDL) fibres; between type 2 fibres, type 2B showed the higher slope of pCa/tension curve. Type 1 fibres from different muscles showed similar calcium sensitivities when containing only the slow set of regulatory proteins and MLC; when both slow and fast isoforms were present, calcium sensitivity shifted toward fast type fibre values. Type 2A fibres from different muscles showed a similar calcium sensitivity, independently of the set (purely fast or mixed) of regulatory proteins and MLC. It is suggested that when both fast and slow isoforms of regulatory proteins and of MLC are present in a muscle fibre, calcium sensitivity is dictated mainly by the fast isoforms

Myosin heavy chain composition of single fibres from normal human muscle

Electrophoretic analysis in the presence of 33% glycerol of purified myosin from normal human muscle shows three distinct protein bands which are identified as type 1, 2B, and 2A myosin heavy chain (MHC) isoforms by affinity-purified polyclonal antibodies. Analysis of MHC of single human muscle fibres shows that human muscles contain a large population of fibres showing the coexistence of type 2A and 2B MHC

Effects of sphingomyelin derivatives on innervated and denervated rat soleus muscle

Present work is aimed at studying the effects of natural bioactive sphingomyelin derivatives, on normal and denervated slow-twitch skeletal muscle. The effects on fibre cross sectional area and myosin heavy chains composition of sphingosine (SPH), sphingosine 1-phosphate (S1P) and sphingosylphosphorylcholine (SPC) were studied. Denervation was bilaterally performed cutting the sciatic nerve at the level of trochanter in adult rats. A group of animals was used as controls. Sphingolipids were continuously released by a mini-osmotic pumps implanted subcutaneously in the scapular region and connected by a catheter to the left (control or denervated) soleus muscles. Supplementation of SPC to adult control soleus muscle, produced significant atrophy of fibres and small changes to fibre type composition. No significant effects of SPH and S1P were found. In contrast, SPH and, to a smaller extent, S1P reduced the atrophy and the slow-to-fast transformation produced by 7-14 days of denervation. Preliminary results indicate that these sphingolipids may exert their action by reducing the overall muscle apoptosis and by activating satellite cells

Role of sphingosylphosphorylcholine on skeletal muscle denervation and regeneration

Present work is aimed at studying the role of sphingosylphosphorylcholine (SPC), a natural bioactive sphingomyelin derivative, on denervation and regeneration of skeletal muscle. SPC is known to participate in growth, proliferation and survival of various cell systems. The experiments were performed in adult rats. SPC was continuously released by a mini-osmotic pumps connected by a catheter to control, denervated or regenerating (after bupivacaine injection) soleus muscles. Supplementation of SPC to adult muscles, produced significant atrophy of fibres and light changes to fibre type composition. In contrast, SPC did not influence denervation atrophy. During regeneration, SPC produced alterations on fibre dimensions and on myosin expression. In conclusion, it appears that SPC exerts distinctive actions depending on definite condition of skeletal muscl