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Purpose This is the first study to examine whether age impacts the response of single muscle fibers to high/low frequency and high/low volume electrical pulse stimulation. We performed in vitro experiments to evaluate the effect of low-frequency high-volume electrical pulse stimulation (EPS) on mechanistic target of rapamycin (mTOR), mitogen-activated protein kinase (MAPK) signaling pathway, and satellite cell activation in singles fibers of young and aged muscles. Methods Isolated single fibers from gastocnemius in 12-wk (n=21) and 72-wk (n=21) old male C57BL/6 mice were divided into four groups: 1) control (Con) received no EPS, 2) low-frequency low-volume EPS (LL), 3) low-frequency high-volume EPS (LH), and 4) high-frequency low-volume EPS (HL) were made to contract using independent EPS protocols. Satellite cell activation and anabolic pathway (mTOR and MAPK signaling) were measured before and after EPS. Results The number of quiescent (Pax7+/Ki67-) and active (Pax7+/Ki67+) satellite cells, myonuclear content and the phosphorylation of 4E-BP1 and ERK were higher in young when compared with old. However, regardless of age, LH and HL EPS significantly increased the number of activate satellite cells (142%, both) and phosphorylation of mTOR (129% and 133%, respectively), p70S6K (133% and 136%, respectively) and 4E-BP1 (140% and 129%, respectively) compared with Con. The protein expression of ERK phosphorylation only increased by LH EPS in both the young and old groups (123% and 125%, respectively). Conclusion Low-frequency high-volume EPS stimulated satellite cell activation and the mTOR signaling pathway in older similar to young muscle.