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The calcium\u2010binding proteins calretinin (CR) and calbindin\u2010D28k (CB) have been widely used to characterize neuronal populations in vertebrate brains. Here, the distribution of the immunoreactivity against CR and CB was investigated in the olfactory organ and brain of <jats:italic>Polypterus senegalus<\/jats:italic> and compared to the distribution of these molecules in other ray\u2010finned fishes. In general, CB\u2010immunoreactive (ir) neurons were less abundant than CR\u2010ir cells. CR immunohistochemistry revealed segregation of CR\u2010ir olfactory receptor neurons in the olfactory mucosa and their bulbar projections. Our results confirmed important differences between pallial regions in terms of CR immunoreactivity of cell populations and afferent fibers. In the habenula, these calcium\u2010binding proteins revealed right\u2010left asymmetry of habenular subpopulations and segregation of their interpeduncular projections. CR immunohistochemistry distinguished among some thalamic, pretectal, and posterior tubercle\u2010derived populations. Abundant CR\u2010ir populations were observed in the midbrain, including the tectum. CR immunoreactivity was also useful for characterizing a putative secondary gustatory\/visceral nucleus in the isthmus, and for distinguishing territories in the primary viscerosensory column and octavolateral region. Comparison of the data obtained within a segmental neuromeric context indicates that some CB\u2010ir and CR\u2010ir populations in polypteriform fishes are shared with other ray\u2010finned fishes, but other positive structures appear to have evolved following the separation between polypterids and other ray\u2010finned fishes. J. Comp. 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