While subtypes 1 and 2 of the ryanodine receptor (RyR) function as intracellular Ca2+ release channels, little is known about the function of the third subtype (RyR-3), first identified in brain. Myocytes from mice homozygous for a targeted mutation in the RyR-1 gene (dyspedic mice) can now be used for a study on the function of RyR-3, which is predominantly expressed in these cells according to our reverse transcription-polymerase chain reaction analysis. We here demonstrate in these myocytes caffeine-, ryanodine- and adenine nucleotide-sensitive Ca(2+)-induced Ca2+ release with approximately 10 times lower sensitivity to Ca2+ than that of RyR-1. Although RyR-3 does not mediate excitation-contraction coupling of the skeletal muscle type, we propose that RyR-3 may induce intracellular Ca2+ release in response to a Ca2+ rise with a high threshold.