Inwardly rectifying potassium channels promote directional sensing during neutrophil chemotaxis

Potassium channels control membrane potential and various physiological processes, including cell migration. However, the specific role of inwardly rectifying potassium channels in immune cell chemotaxis remains unknown. Here, we demonstrate that inwardly rectifying potassium channels, particularly Kir7.1 (Kcnj13), maintain the resting membrane potential and are crucial for directional sensing during neutrophil chemotaxis. Blocking or knocking out Kir in neutrophils disrupted their ability to sense direction toward different chemoattractants in multiple models. Using genetically encoded voltage indicators, we observed oscillating hyperpolarization during tail retraction in zebrafish neutrophils, with Kir7.1 required for depolarization toward the chemokine source. Focal depolarization via optogenetics biased pseudopod selection and triggered new protrusions, which depended on Gα signaling. Global hyperpolarization caused neutrophils to stall migration. Additionally, Kir influences GPCR signaling activation in dHL-60 cells. This research introduces membrane potential as a key component of the complex feedforward mechanism that links the adaptive and excitable networks necessary to guide immune cells in challenging tissue environments.