The effects of low frequency electrical stimulation on satellite cell activity in rat skeletal muscle during hindlimb suspension

Bao-Ting Zhang, Simon S Yeung, Yue Liu, Hong-Hui Wang, Yu-Min Wan, Shu-Kuan Ling, Hong-Yu Zhang, Ying-Hui Li, Ella W Yeung, Bao-Ting Zhang, Simon S Yeung, Yue Liu, Hong-Hui Wang, Yu-Min Wan, Shu-Kuan Ling, Hong-Yu Zhang, Ying-Hui Li, Ella W Yeung

Abstract

Background: The ability of skeletal muscle to grow and regenerate is dependent on resident stem cells called satellite cells. It has been shown that chronic hindlimb unloading downregulates the satellite cell activity. This study investigated the role of low-frequency electrical stimulation on satellite cell activity during a 28 d hindlimb suspension in rats.

Results: Mechanical unloading resulted in a 44% reduction in the myofiber cross-sectional area as well as a 29% and 34% reduction in the number of myonuclei and myonuclear domains, respectively, in the soleus muscles (P < 0.001 vs the weight-bearing control). The number of quiescent (M-cadherin(+)), proliferating (BrdU(+) and myoD(+)), and differentiated (myogenin(+)) satellite cells was also reduced by 48-57% compared to the weight-bearing animals (P < 0.01 for all). Daily application of electrical stimulation (2 × 3 h at a 20 Hz frequency) partially attenuated the reduction of the fiber cross-sectional area, satellite cell activity, and myonuclear domain (P < 0.05 for all). Extensor digitorum longus muscles were not significantly altered by hindlimb unloading.

Conclusion: This study shows that electrical stimulation partially attenuated the decrease in muscle size and satellite cells during hindlimb unloading. The causal relationship between satellite cell activation and electrical stimulation remain to be established.

Figures

Figure 1
Figure 1
Muscle fiber size. (A) Cross-sections of soleus muscle stained with H & E from WB, HU, and HU-ES groups. Scale bar = 100 μm. Mean fiber cross-sectional area (μm2) of soleus (B) and EDL (C) muscles in the WB, HU, and HU-ES groups. *** P < 0.001, significant difference between WB and HU groups; # P < 0.05, significant difference between HU and HU-ES groups. n = 4. Data are means ± SEM.
Figure 2
Figure 2
Myonuclear number and domain size. (A, B) Numbers of myonuclei per millimeter fiber length, and (C, D) Myonuclear domain size for WB, HU, and HU-ES groups. *** P < 0.001, significant difference between WB and HU groups; # P < 0.05, significant difference between HU and HU-ES groups. n = 4. Data are means ± SEM.
Figure 3
Figure 3
Changes in the markers of satellite cells. Representative cross-sections of soleus muscle immunostained for M-cadherin, BrdU, myoD, and myogenin (green, arrows), respectively. The two columns on the left are DIC images. The two columns on the right show dystrophin immunolabeling (red) to identify fiber profiles and nuclear staining with DAPI (blue). Scale bar = 20 μm.
Figure 4
Figure 4
Quantification of satellite cells labeled positive in soleus muscles. Number of M-cadherin+ cells or BrdU+, myoD+, and myogenin+ nuclei per 100 fibers in cross sections of the soleus muscles. ** P < 0.01, significant difference between WB and HU groups; # P < 0.05, significant difference between HU and HU-ES groups. n = 6. Data are means ± SEM.

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