Using a single, mechanically skinned fiber approach we tested the hypothesis that denervation (0-50-d) of skeletal muscles that do not overlap in fiber type composition (extensor digitorum longus, EDL, and soleus, SOL, muscles of Long-Evans Hooded rats) leads to development of different fiber phenotypes. Denervation (50-d) was accompanied by (i) a marked increase in the proportion of hybrid IIB/D fibers (EDL) and I/IIA fibers (SOL) from 30% to >75% in both muscles, and a corresponding decrease in the proportion of pure fibers expressing only one myosin heavy chain (MHC) isoform; (ii) complex, muscle- and fiber-type specific changes in sarcoplasmic reticulum Ca²⁺-loading level at physiological pCa (-log₁₀[Ca²⁺]) 7.1, with EDL fibers displaying more consistent changes than SOL fibers; (iii) decrease by ~50% in specific force of all fiber types; (iv) decrease in sensitivity to Ca²⁺, particularly for SOL fibers (by ~40%); (v) decrease in the maximum steepness of the force-pCa curves, particularly for the hybrid I/IIA SOL fibers (by 35%) and (vi) increased occurrence of biphasic behaviour with respect to Sr²⁺ activation in SOL fibers, indicating the presence of both slow- and fast-troponin C isoforms. No fiber types common to the two muscles were detected at any time points (7-, 21- and 50-d) post-denervation. The results provide strong evidence that not only neural factors, but also the intrinsic properties of a muscle fiber influence the structural and functional properties of a particular muscle cell and explain important functional changes induced by denervation at both whole muscle and single cell levels.