1. Ferret papillary muscles were isolated and injected with aequorin to measure intracellular Ca2+ concentration [( Ca2+]i). Developed tension and [Ca2+]i were measured in response to length changes. 2. A maintained reduction in muscle length produced an immediate decrease in developed tension followed by slow decline over 10‐20 min. This slow decline in tension was accompanied by a slow decline in the amplitude of the systolic [Ca2+]i rise (the Ca2+ transient). The immediate decrease in tension was accompanied by a prolongation of the Ca2+ transient and an abbreviation of the twitch. 3. Repeated reductions in muscle length timed to occur only during the period of contraction (systolic shortening) produced an immediate decrease of developed tension but the subsequent slow decline was substantially smaller. The slow decline in the amplitude of the Ca2+ transients was also smaller. The prolongation of the Ca2+ transient and abbreviation of the twitch were similar to those observed with a maintained reduction of length. 4. Repeated reductions in muscle length during the period between contractions (diastolic shortening) did not produce the immediate decrease of tension but the slow decline of tension was present. The slow decline in the amplitude of the Ca2+ transients was also present. However no change in the duration of the Ca2+ transient or the twitch was present under these conditions. 5. These results suggest that diastolic muscle length can influence the amplitude of the Ca2+ transients achieved during systole. This conclusion was confirmed by experiments in which the recovery of tension and Ca2+ transients was observed after periods of rest. Both developed tension and Ca2+ transients on recovery from a rest were reduced when the rest occurred at a short length in comparison with a long length. 6. We suggest that muscle length influences resting [Ca2+]i and this in turn affects the Ca2+ transients and developed tension.