It is poorly understood how mechanical stimuli influence in-vivo myocardial remodeling during chronic hemodynamic overload. Combined quantitation of ventricular mechanics and expression of key proteins involved in mechanotransduction can improve fundamental understanding. Methods: Adult anesthetized dogs (n=20) were studied at sinus rhythm (SR) and 0, 3, 10 and 35 days of complete atrioventricular block (AVB). Serial left-ventricular (LV) myofiber mechanics were measured. Repeated LV biopsies were analyzed for mRNA and/or protein expression of _1D integrin, melusin, Akt, glycogen-synthase-kinase-3 (GSK3), muscle LIM protein (MLP), four-and-a-half LIM protein 2 (fhl2), desmin, and calpain. Results: Upon AVB, increased ejection strain (0.29±0.01 vs. 0.13±0.02, SR) and end-diastolic stress (4.8±1.1 vs. 2.7±0.4 kPa) dominated mechanical changes. Brain natriuretic peptide plasma levels were correspondingly high (33±4 vs. 19±1 pg/mL, SR). _1D-Integrin protein expression increased chronically after AVB. Melusin was temporarily overexpressed (+33±9%, 3 days AVB vs. SR), followed by elevated ratios of phosphorylated (P)-Akt / Akt and P-GSK3 / GSK3 (+26±6% and +30±8% at 10 days AVB vs. SR). These changes corresponded to peak hypertrophic growth at 3 to 10 days. MLP increased gradually to maxima at chronic AVB (+36±7%). In contrast, fhl2 (-22±3%, 3 days) and desmin (-30±9%, 10 days AVB) transiently declined but recovered at chronic AVB. Calpain protein expression remained unaltered. Conclusions: Volume overload after AVB causes a transient compromise of cytoskeletal integrity based, at least partly, on transcriptional downregulation. Subsequent cytoskeletal reorganization coincides with the upregulation of melusin, P-Akt, P-GSK3 and MLP, indicating a strong drive to compensated hypertrophy.