Eccentric contractions (EC) are known to result in muscle hypertrophy, potentially through activation of the Akt-mTOR-p70S6k signaling pathway. Previous work has also demonstrated that EC result in the opening of stretchactivated channels (SAC) and inhibition of these channels resulted in an attenuation of the EC to induce muscle hypertrophy. The purpose of this study was to test the hypothesis that a known intracellular pathway directly associated with muscle hypertrophy is coupled to the opening of stretch activated ion channels (SAC). Specifically we measured the activation of the Akt, GSK-3, p70S6k, and ribosomal protein S6 following a single bout of EC in the rat tibialis anterior (TA) muscle. The TA muscles preformed four sets of six repetitions of eccentric contractions. In-vivo blockade of SAC was performed by a continuous oral treatment with streptomycin in the drinking water (4g/I) or by i.v. infusion of Gadolinium (Gd³⁺) 80µmol/kg. EC increased the degree of Akt and p70^s6k phosphorylation in the TA muscle, while in animals where SAC had been inhibited there was a reduced capacity for EC to induce Akt or p70^s6k phosphorylation. Accompanying this reduced activation of Akt and p70^s6k, was a failure to phosphorylate GSK-3 or S6 when SAC were inhibited. The results from these data indicate the necessity of functional SAC for the complete activation of Akt and p70S6k pathway in response to EC.