Calsequestrin (CAL) is a calcium-binding protein whose primary function is thought to involve sequestration of calcium in the muscle sarcoplasmic reticulum (SR). Little is known about the mechanisms regulating CAL expression, or about the role of this protein in muscle development. In addition, CAL may regulate calcium localization in some nonmuscle cells. We have identified an avian calsequestrin homolog. The predicted amino acid sequence of the avian CAL, first described as a laminin binding protein, and named aspartactin, is 70-80% identical to mammalian CAL sequences. We have used affinity-purified antibodies and cDNA probes to investigate expression in developing and adult chicken tissues. In adult chickens, the avian CAL homolog was expressed in slow and fast twitch skeletal muscle as well as in cardiac muscle. Surprisingly high levels of CAL protein were also detected in cerebellum. During development, CAL mRNA and protein were detected in Embryonic Day 5 (E-5) limb primordia, well before the initiation of myoblast fusion. In leg skeletal muscle, CAL protein and mRNA increase approximately 10-fold from E-8 to E-18 with a time course that just precedes myoblast fusion. This early expression pattern was also observed in cultured chicken pectoral myoblasts, and appears to be regulated at the level of mRNA abundance. The developmental profile of CAL expression is compared to that of other muscle proteins and possible additional functions of CAL are discussed.