Fig. 5: CLP257 enhances KCC2 membrane expression in rats with peripheral nerve injury (PNI).
From: Enhancing neuronal chloride extrusion rescues α2/α3 GABAA-mediated analgesia in neuropathic pain
a Confocal laser scanning microscopy (CLSM) images showing KCC2 immunostaining in PNI vs. sham animals. Insets show examples of KCC2 profiles of magnified neurons within these two confocal acquisitions (Y-scale bar = 500 i.u.; horizontal dashed lines are 3 pixel-thick and correspond to a selected KCC2 profile and the intensity axis origin). b Averaged intensity KCC2 profiles of visually identified sham and PNI neurons across their plasma membrane. Sham (Cyan) n = 77 neurons in 8 rats; PNI (red) n = 190 neurons in 8 rats. c CLSM images of KCC2 immunostaining in PNI vehicle vs. PNI CLP257 i.t. injected rats. Insets show examples of KCC2 profiles of magnified neurons within these two confocal acquisitions. d Averaged intensity KCC2 profiles in PNI + vehicle (red) n = 268 neurons in 12 rats and in PNI + CLP257 (orange) n = 310 neurons in 17 rats. e Method of global index intensity analysis to determine KCC2 staining at the membrane. f Global index KCC2 analysis of the averaged KCC2 immunostaining pixel intensity at the membrane and in the intracellular space in PNIs vs. shams. g Pixel intensity of KCC2 immunostaining in PNIs where vehicle or 40 mg kg−1 of CLP257 was i.t. injected 2 h prior to tissue fixation. Dots in Fig. 5f, g represent single rats. h Ultrastructural immunostaining of KCC2 of a dorsal horn spinal cord neuron in a naive rat; subcellular compartments are also displayed. i Magnified image showing the subcellular distribution of KCC2. j Higher magnification of the area delimited in i. Note the membrane KCC2 enriched on both sides of an inhibitory synaptic connection and KCC2 in endosomes. Syn.: synapse; (*P < 0.05; **P < 0.01). Source data is available as a Source Data file.
