Pannexin 1 Channels Link Chemoattractant Receptor Signaling to Local Excitation and Global Inhibition Responses at the Front and Back of Polarized Neutrophils

Neutrophil chemotaxis requires excitatory signals at the front and inhibitory signals at the back of cells, which regulate cell migration in a chemotactic gradient field. We have previously shown that ATP release via pannexin 1 ( PANX1 ) channels and autocrine stimulation of P2Y 2 receptors contribute to the excitatory signals at the front. Here we show that PANX1 also contributes to the inhibitory signals at the back, namely by providing the ligand for A 2A adenosine receptors. In resting neutrophils, we found that A 2A receptors are uniformly distributed across the cell surface. In polarized cells, A 2A receptors redistributed to the back where their stimulation triggered intracellular cAMP accumulation and protein kinase A (PKA) activation, which blocked chemoattractant receptor signaling. Inhibition of PANX1 blocked A 2A receptor stimulation and cAMP accumulation in response to formyl peptide receptor stimulation. Treatments that blocked endogenous A 2A receptor signaling impaired the polarization and migration of neutrophils in a chemotactic gradient field and resulted in enhanced ERK and p38 MAPK signaling in response to formyl peptide receptor stimulation. These findings suggest that chemoattractant receptors require PANX1 to trigger excitatory and inhibitory signals that synergize to fine-tune chemotactic responses at the front and back of neutrophils. PANX1 channels thus link local excitatory signals to the global inhibitory signals that orchestrate chemotaxis of neutrophils in gradient fields. Background: Chemotaxis requires excitatory and inhibitory signals at the front and back of cells. PANX1 and P2Y 2 receptors provide local excitation at the front. Results: We found that PANX1 triggers A 2A receptor activation and inhibits chemotactic signaling at the back. Conclusion: PANX1 thus integrates excitatory and inhibitory signals that regulate chemotaxis at the front and back of cells. Significance: These findings suggest new strategies to modulate neutrophil chemotaxis.