Nonlinear summation of contractions is studied in single barnacle (Balanus nubilus) muscle fibres, loaded with the photoprotein aequorin. The results indicate that nonlinear summation of aequorin transients is indeed present and for short interpulse intervals (25-250 ms), a more-than-linear summation of transients, which suggest an increase of the cytosolic Ca2+ concentration in the second response, is observed. This augmented Ca2+ concentration is not merely due to summation with the preceding conditioning transient, but to an enlargement of the second transient in its own right. Furthermore, the enlargement of the second Ca2+ response is not the result of prolonged release, or slowing of re-uptake by intracellular organelles. On the contrary, Ca2+ release is found to be enhanced and for short depolarizations (20 ms), its time to half re-uptake is reduced. The intensified Ca2+ release, triggered by the second standard depolarization, is related to the level of cytosolic Ca2+ concentration reached in the conditioning response and, for example, appears to be larger in the presence of Dantrolene-sodium, which is known to reduce Ca2+ movements in a single twitch. It is concluded that contractile potentiation observed during nonlinear summation of contractions, is associated with a potentiation of intracellular Ca2+ movements, which interact to regulate the cytosolic Ca2+ concentration during contraction.