Real-time functional analysis of Hv1 channel in neutrophils: a new approach from zebrafish model
| Autor: | Takafumi Kawai, Yasushi Okamura, Adisorn Ratanayotha |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Proton channel Time Factors Physiology Neutrophils Phagocytosis Ion Channels Membrane Potentials Animals Genetically Modified 03 medical and health sciences 0302 clinical medicine Immune system Physiology (medical) Phagosomes Animals Calcium Signaling Zebrafish chemistry.chemical_classification Reactive oxygen species biology Functional analysis Zebrafish Proteins biology.organism_classification In vitro Immunity Innate Cell biology 030104 developmental biology chemistry Reactive Oxygen Species Ion Channel Gating 030217 neurology & neurosurgery Communication channel |
| Zdroj: | American journal of physiology. Regulatory, integrative and comparative physiology. 316(6) |
| ISSN: | 1522-1490 |
| Popis: | Voltage-gated proton channel (Hv1) has been studied in various immune cells, including neutrophils. However, most studies have taken an in vitro approach using isolated cells or primary cultured cells of mammals; therefore, limited evidence is available on the function of Hv1 in a physiological context. In this study, we have developed the in vivo system that enables real-time functional analysis of Hv1 using zebrafish embryos ( Danio rerio). Hvcn1-deficiency ( hvcn1−/−) in zebrafish completely abolished voltage-gated proton current, which is typically observed in wild-type neutrophils. Importantly, hvcn1-deficiency significantly reduced reactive oxygen species production and calcium response of zebrafish neutrophils, comparable to the results observed in mammalian models. These findings verify zebrafish Hv1 (DrHv1) as the primary contributor for native Hv1-derived proton current in neutrophils and suggest the conserved function of Hv1 in the immune cells across vertebrate animals. Taking advantage of Hv1 zebrafish model, we compared real-time behaviors of neutrophils between wild-type and hvcn1−/− zebrafish in response to tissue injury and acute bacterial infection. Notably, we observed a significant increase in the number of phagosomes in hvcn1−/− neutrophils, raising a possible link between Hv1 and phagosomal maturation. Furthermore, survival analysis of zebrafish larvae potentially supports a protective role of Hv1 in the innate immune response against systemic bacterial infection. This study represents the influence of Hv1 on neutrophil behaviors and highlights the benefits of in vivo approach toward the understanding of Hv1 in a physiological context. |
| Databáze: | OpenAIRE |
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