We examined the effects of post-stimulus length changes on voltage-clamped, aequorin-injected single muscle fibers from the barnacle Balanus nubilus. Extra light (extra calcium) is seen when the fiber is allowed to shorten (a small percentage) during the declining phase of the calcium transient. The opposite is observed when the fiber is stretched. Increasing the extent of shortening increases the amount of extra calcium, as does decreasing the temperature. The extra calcium probably comes from the myofilaments and not from the sarcoplasmic reticulum because (a) there is a strong correlation between the extra calcium and the level of activation; (b) there is a strong correlation between the extra calcium and the amount of force redeveloped after a length change; and (c) the time course of the appearance of the extra calcium is intermediate between that of the free calcium concentration and that of force. We suggest (a) that the calcium binding to the activating myofibrillar proteins is sensitive to muscle length or muscle force, and (b) that there is a pool of bound calcium (activating calcium) that waxes and wanes with a time course intermediate between the free calcium concentration and force.

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