Previous molecular studies indicated that antimicrobial peptides in lizard are expressed in the skin and tongue among other epithelial organs. The present ultrastructural immunogold study aimed to detect the specific location of three broadly expressed antimicrobial peptides in the tongue of the lizard Anolis carolinensis. The immunocytochemical study indicated that beta-defensin-15, the likely main defensin of granulocytes and skin, is poorly expressed in some dense and medium-dense granules of glandular cells of the papillated tongue. Conversely beta-defensin-27 appears highly expressed in numerous pale and cribriform dense granules of glandular cells and is also secreted on the tongue surface. The immunostaining for cathelicidin-1 indicated a variable but however positive immunolabeling in numerous granules in the tongue glands, suggesting that this antimicrobial peptide previously found on the epidermal surface is also present in the tongue secretions and participates to the formation of the anti-microbial oral barrier. The study suggests that among the numerous beta-defensins and cathelicidins identified in the genome of this lizard is present a specific distribution of different peptide subtypes in various body regions, including the tongue, and that these peptides contribute to the formation of local antimicrobial barriers.