Regulation of melanin-based pigmentation is complex, involving multiple genes. Because different genes can contribute to the same pigmentation phenotype, the genes identified in model organisms may not necessarily apply to wild species. In the barn owl (Tyto alba), ventral plumage colour ranges from white to rufous, with genetic variation in the melanocortin 1 receptor gene (MC1R) accounting for at least a third of this variation. In the present study, we used transcriptomic data to compare the gene expression profiles of growing feathers from nestlings with different MC1R genotypes. We identified 21 differentially expressed genes, nine of which are involved in melanogenesis, while seven are related to neurotransmitter function or synaptic activity. With the exception of CALB1, all of the differentially expressed genes were upregulated in rufous owls compared to white barn owls. To the best of our knowledge, this study is the first to link melanin production with neurotransmitter-related genes, and we discuss possible evolutionary explanations for this connection.