Role of the Intracellular Sodium Homeostasis in Chemotaxis of Activated Murine Neutrophils

The importance of the intracellular Ca²⁺ concentration ([Ca²⁺]i) in neutrophil function has been intensely studied. However, the role of the intracellular Na⁺ concentration ([Na⁺]i) which is closely linked to the intracellular Ca²⁺ regulation has been largely overlooked. The [Na⁺]i is regulated by Na⁺ transport proteins such as the Na⁺/Ca²⁺-exchanger (NCX1), Na⁺/K⁺-ATPase, and Na⁺-permeable, transient receptor potential melastatin 2 (TRPM2) channel. Stimulating with either N-formylmethionine-leucyl-phenylalanine (fMLF) or complement protein C5a causes distinct changes of the [Na⁺]i. fMLF induces a sustained increase of [Na⁺]i, surprisingly, reaching higher values in TRPM2-/- neutrophils. This outcome is unexpected and remains unexplained. In both genotypes, C5a elicits only a transient rise of the [Na⁺]i. The difference in [Na⁺]i measured at t = 10 min after stimulation is inversely related to neutrophil chemotaxis. Neutrophil chemotaxis is more efficient in C5a than in an fMLF gradient. Moreover, lowering the extracellular Na⁺ concentration from 140 to 72 mM improves chemotaxis of WT but not of TRPM2-/- neutrophils. Increasing the [Na⁺]i by inhibiting the Na⁺/K⁺-ATPase results in disrupted chemotaxis. This is most likely due to the impact of the altered Na⁺ homeostasis and presumably NCX1 function whose expression was shown by means of qPCR and which critically relies on proper extra- to intracellular Na⁺ concentration gradients. Increasing the [Na⁺]i by a few mmol/l may suffice to switch its transport mode from forward (Ca²⁺-efflux) to reverse (Ca²⁺-influx) mode. The role of NCX1 in neutrophil chemotaxis is corroborated by its blocker, which also causes a complete inhibition of chemotaxis.