Serotonin stimulates proliferation of ionocytes via the 5-HT2A receptor in zebrafish larvae

Osmoregulation is an essential process in all living organisms. For aquatic organisms, such as freshwater fishes whose natural environment is hyperosmotic, specialized cells, called ionocytes, are present in the skin during developmental stages and contribute to the maintenance of osmotic homeostasis. Such cells are known to proliferate in response to osmotic stress, but the molecular mechanism by which that process is regulated remains poorly characterized. In this study, using immunohistochemistry and confocal microscopy, we demonstrate that cutaneous ionocytes in developing zebrafish ( Danio rerio ) express serotonin 2A (5-HT2A) receptors by co-labeling with other known ionocyte markers, such as the Na ⁺ /K ⁺ -ATPase, Concanavalin A and Mitotracker. Furthermore, by quantifying ionocyte number through early stages of development, we implicate 5-HT2A receptors in initiating ionocyte proliferation. Exposure of zebrafish embryos and larvae to acidic pH, or exogenous 5-HT, increased the number of cutaneous ionocytes. The effects of both stimuli were abolished in the presence of the 5-HT2A receptor-specific antagonist, ketanserin. Moreover, activation of 5-HT2A receptors led to increased detection of ionocytes with phosphorylated extracellular signal-related kinase (ERK), a key regulator of cell division and differentiation linked with 5-HT2A. We used tetrabenazine, an inhibitor of vesicular monoamine transporter 2 (vmat2) and 5-HT storage, to deplete potential sources of 5-HT. Tetrabenazine treatment in fish exposed to acidic pH reduced ionocyte proliferation, implicating an endogenous source of 5-HT in regulation of ionocyte populations.These results demonstrate the importance of a pathway initiated by 5-HT2A activation that regulates ionocyte proliferation in developing zebrafish exposed to osmotic stress.