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Knowledge of the histochemical substrates of cellular and neurovascular connections in the gingiva is essential in order to understand the initial mechanisms of inflammation in the periodontium. Since the localization of the neuroendocrine peptide galanin in the gingiva is still unclear, we used immunohistochemical, in situ hybridization and immunoblot techniques to assess the localization of galanin in the gingiva of rat molars. Galanin-immunoreactive nerve fibers were located around blood vessels in the lamina propria, beneath the epithelium, in the epithelial–proprial junction and in the basal layer of the epithelium. Galanin was highly expressed in the suprabasal keratinocytes of the gingival epithelium. The localization of galanin in gingival nerve fibers and the expression of galanin in keratinocytes of the gingival epithelium indicate that galanin may be a possible regulator of different cellular functions in the gingiva.