Many immunoglobulin (Ig)-superfamily cell adhesion molecules influence skeletal muscle formation. In Drosophila, dumbfounded (duf/kirre), irreC, sticks and stones and hibris encode related Ig-family proteins expressed in subsets of neurons and muscle precursor cells. The family mediates cell migration, axon guidance and fusion of myoblasts. Despite the importance of these genes in invertebrate myogenesis, no obvious functional parallels are known in vertebrate myogenesis. Here we investigate the gene expression pattern and phylogenetic and protein-structural relationships of the duf-related molecules neurolin and neurolin-like cell adhesion molecule (NLCAM), members of the activated leukocyte cell adhesion molecule (ALCAM) sub-family of Ig-molecules. These proteins are among the closest to Duf/Kirre by sequence. During zebrafish development, neurolin is expressed in subsets of somite and muscle cells, heart and numerous sites of neuronal maturation. The new ALCAM-family member, NLCAM, appears to have arisen by duplication of neurolin/ALCAM. NLCAM is expressed widely during gastrulation, particularly in the nascent neural plate, but later becomes predominantly expressed in sites of muscle and nerve maturation and in the fin fold. The expression of each gene is often in groups of cells in similar parts of the embryo; for example, in the region of Rohon Beard neurons, trigeminal ganglion and fusing fast and migrating slow muscle fibres. However, expression can also be distinct and dynamic; for example, muscle pioneer fibres express neurolin but not NLCAM at high level. Both molecules are expressed in subsets of muscle precursors at times prior to fusion.