Rotator cuff (RC) tears present a treatment challenge due to muscle atrophy and degeneration, fatty infiltration, and fibrosis. The purpose of this study was to generate a high time-resolution model of RC tear in rabbits and to characterize the progression of architectural and histological changes. Thirty-five female New Zealand White rabbits (age: 6 months) underwent left supraspinatus tenotomy. Five rabbits were used to evaluate immediate muscle architectural changes. The remaining 30 rabbits underwent right shoulder sham surgery and sacrifice at 1, 2, 4, 8, or 16 weeks. Histology was used to quantify muscle fiber cross-sectional area (CSA), muscle degeneration and regeneration, and fat localized to inter- versus intrafascicular regions. Muscle fiber CSA decreased by 26.5% compared to sham at 16 weeks (effect of treatment, p < 0.0001). Muscle degeneration increased after tenotomy (effect of treatment, p = 0.0006) without any change in regeneration. Collagen and fat content increased by 4 weeks and persisted through 16 weeks. Interfascicular fat was increased at all time points, but intrafascicular fat was increased only at 1, 4, and 16 weeks posttenotomy. Intrafascicular fat adjacent to degenerating muscle fibers increased as well (effect of treatment, p < 0.0001; effect of time, p = 0.0102). Statement of clinical relevance: Rabbit supraspinatus tenotomy recapitulates key features of the pathophysiology of human RC tears, including muscle atrophy and degeneration, lack of regeneration, fat accumulation, and fibrosis.