Glucocorticoid hormones are important intermediates between an organism and its environment. They enable an organism to adjust its behavioural and physiological processes in response to environmental changes by binding to mineralocorticoid receptors (MR) and glucocorticoid receptors (GR) expressed in many tissues, including the integument. The regulation of glucocorticoids co-varies with melanin-based colouration in numerous species, an association that might result from pleiotropic effects of genes in the melanocortin system and evolve within a signalling context. Most studies have focused on the circulating levels of glucocorticoids disregarding the receptors that mediate their action, and that might partly account for the covariation between the regulation of stress and melanin-based colouration. We investigated the association of the expression levels of GR and MR genes with melanin-based colouration in the growing feathers of nestling barn owls (Tyto alba). We also explored the association between GR and MR expression levels and the expression of genes related to the melanocortin system and melanogenesis to better understand the origin of the link between the expression of receptors to which corticosterone binds and melanin-based colouration. Nestling barn owls displaying larger eumelanic black feather spots expressed GR and MR at lower levels than smaller-spotted individuals. However, we found that the expression of the GR and MR genes was positively rather than negatively correlated with the expression of genes involved in the deposition of melanin pigments at the time we sampled the nestlings. This provides mixed evidence of the association between melanin-based traits and MR and GR gene expression. The finding that the expression of GR and MR was positively associated with the expression of the PCSK2 gene (encoding one of the protein convertase responsible for the production of hormone peptide ACTH and α-MSH) suggests that the melanocortin system may be implicated in the establishment of the covariation between melanin-based colour and the expression of receptors to which glucocorticoids bind. However, further studies investigating the expression of melanin-based traits with stress-related endpoints at different time points of feather development will be necessary to understand better the proximate mechanism linking melanin-based traits with stress.