AbstractMuscle‐tendon interactions are important for the establishment of a functional musculoskeletal system. Fgf4 and Fgf8 are expressed in muscle and tendon boundary regions during limb development, suggesting a potential role for Fgf signaling pathway in muscle and tendon interactions. We have examined the expression of Fgf syn‐expression group components during muscle and tendon formation of vertebrate limb development. We observed that the transcriptional effector of Fgf signaling, Pea3, and the modulators of Fgf signal, Sprouty1 and 2, were expressed in muscles and tendons and that their expression was enhanced at the myotendinous junctions in chick and mouse limbs. Analysis of Pea3 and Sprouty gene expression in muscleless limbs of Pax3 mutant mice indicated a major expression in muscles but also revealed that the Pea3 and Sprouty expression in tendons depended on muscles. Finally, our data showed that Fgf4 positively regulated Pea3, Sprouty1, and 2 expression in chick limb mesenchyme. Developmental Dynamics 238:1195–1206, 2009. © 2009 Wiley‐Liss, Inc.