Distribution and functionality of Arch-EGFP pumps in the peripheral sensory pathways of Nav1.8-Arch+ mice. Distribution and functionality of Arch-EGFP.

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Distribution and functionality of Arch-EGFP pumps in the peripheral sensory pathways of Nav1.8-Arch+ mice. Distribution and functionality of Arch-EGFP pumps in the peripheral sensory pathways of Nav1.8-Arch+ mice. Confocal micrographs showing the fluorescence of Arch-EGFP (green), CGRP immunostaining (blue), P2X3 labeling (red), and the merge. A, Arch-EGFP colocalizes with either P2X3 or CGRP in dorsal root ganglia neurons, validating its selective expression in nociceptors. B, Arch-EGFP fluorescence overlaps with CGRP and P2X3 labeling in laminae I and II of the dorsal horn of spinal cord. C, Arch-EGFP colocalizes with CGRP and P2X3 in free nerve endings in the lower and upper dermis of glabrous skin. Insets are at lower magnification (20×). Scale bars, 50 μm. D, Representative yellow (589 nm) light-induced outward photocurrent and membrane hyperpolarization in a DRG neuron. Magnification shows an inward deflection in the voltage-clamp trace (arrow), illustrating a proton-mediated ASIC-like current. This inward current did not translate into a depolarization in the current-clamp trace. E, Arch-mediated blockade of electrically induced action potentials (10 ms, 0.4 nA current injection) in DRG neurons. Optical stimulation was continuous (intensity, 0.83 mW/mm2). Vh = −60 mV in voltage-clamp configuration; and the resting membrane potential was −62.23 ± 2.92 mV in current-clamp configuration (n = 8–9 cells). Ihab Daou et al. eneuro 2016;3:ENEURO.0140-15.2016 ©2016 by Society for Neuroscience