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Abstract Chemical signalling is an essential component of the communication system of lizards, and epidermal glands are responsible for producing semiochemicals that regulate many behavioural interactions. Two types of epidermal glands have been previously described for lizards: follicular and generation glands. Generation glands are characterized by the aggregation of novel glandular cell types in the epithelium and the lack of a lumen or external pore. Despite the fact that several subtypes of generation glands have been recognized over the years, the morphology, taxonomic distribution, function and evolutionary origins of generation glands remain nearly unexplored in Neotropical clades. Here, we describe a novel escutcheon-type generation gland (‘α-gland’) for lizards of the South American family Tropiduridae, characterize its structural and ultrastructural organization, and study the homology of the constituent parts in a phylogenetic framework. The α-glands emerged in the ancestor of Eurolophosaurus, Plica, Strobilurus, Tropidurus and Uracentron, and are found in at least 39 species with diverse ecological habits. We preliminarily analysed the protein profile of α-glands and discovered differential expression of protein components between sexes. Our investigations change the general view about epidermal gland homology, leading us to argue that generation and follicular glands are possibly more closely related functionally and evolutionarily than previously thought.