The contribution of Ca2+ released from the sarcoplasmic reticulum (SR) to smooth muscle contractile activation remains poorly understood. By simultaneously monitoring cytosolic [Ca2+] ([Ca2+]i) and force in isolated rat uterine smooth muscle, we report the influence of SR Ca2+ release on contractility during conditions (a) of altered SR Ca2+ homeostasis and (b) where the only source of activating Ca2+ was derived from the SR. 2. In myometria of non-pregnant rats, ryanodine (1-50 microM), a modulator of calcium-induced calcium release (CICR), had no effect on the spontaneous [Ca2+]i or force transients. However, depletion of SR Ca2+ by inhibiting the SR Ca2+-ATPase (with cyclopiazonic acid (CPA), 20 microM) resulted in an enhancement of spontaneous [Ca2+]i and force transients. 3. In myometria of pregnant rats, although ryanodine had no effect in 40% of tissues studied it produced a small but significant enhancement of the integrated spontaneous [Ca2+]i and force transient in 60% of cases. The potentiating effects of CPA were enhanced in myometria of pregnant rats compared with non-pregnant rats, often resulting in maintained [Ca2+]i increases and contraction. 4. In zero external Ca2+, agonist-induced SR Ca2+ release resulted in transient increases in [Ca2+]i and force. The magnitude of these agonist-induced [Ca2+]i and force changes were significantly enhanced in myometria of pregnant rats. No evidence for agonist-induced Ca2+-independent force production was observed. 5. These results indicate that CICR plays little role in SR Ca2+ release from the myometrium, and that there are gestational-dependent alterations in the ability of SR Ca2+ mobilization to contribute to contractile activation. The implications of these findings for the co-ordination of myometrial [Ca2+]i signalling and contractility are discussed.