Molecular and functional characterization of the voltage-gated proton channel in zebrafish neutrophils

Voltage-gated proton channels (Hv1/VSOP) are expressed in various cells types, including phagocytes, and are involved in diverse physiological processes. Although hvcn1, the gene encoding Hv1, has been identified across a wide range of species, most of the knowledge about its physiological function and expression profile is limited to mammals. In this study, we investigated the basic properties of DrHv1, the Hv1 ortholog in zebrafish (Danio rerio) which is an excellent animal model owing to the transparency, as well as its functional expression in native cells. Electrophysiological analysis using a heterologous expression system confirmed the properties of a voltage-gated proton channel are conserved in DrHv1 with differences in threshold and activation kinetics as compared to mouse (Mus musculus) Hv1 (mHv1). RT-PCR analysis revealed that hvcn1 is expressed in zebrafish neutrophils, as is the case in mammals. Subsequent electrophysiological analysis confirmed the functional expression of DrHv1 in zebrafish neutrophils, which suggests Hv1 function in phagocytes is conserved among vertebrates. We also found that DrHv1 is comparatively resistant to extracellular Zn²⁺, which is a potent inhibitor of mammalian Hv1, and this phenomenon appears to reflect variation in the Zn²⁺-coordinating residue (histidine) within the extracellular linker region in mammalian Hv1. Notably, the serum Zn²⁺ concentration is much higher in zebrafish than in mouse, raising the possibility that Zn²⁺ sensitivity was acquired in accordance with a change in the serum Zn²⁺ concentration. This study highlights the biological variation and importance of Hv1 in different animal species.