Figure 6.

Electrophysiological characteristics of CFP+ FABP7− cells from the DRG of hGFAP-CFP mice. (A) Representative images of dissociated DRG culture from hGFAP-CFP (green) mice at 7 days after SNI, with immunofluorescence staining against SGC marker FABP7 (red). Dotted circle delineates the neuronal soma. All observed CFP+ cells that are detached from neuronal soma are FABP7−. Potassium current density of detached CFP+ cells in dissociated culture of contralateral or ipsilateral L3, L4 and L5 DRG, at 2 days (B) and 7 days (C) after SNI. The 4AP at 1 mM was used to block K_v currents (two-way ANOVA: B. Interaction Group F(2, 722) = 48.37 p < 0.0001 with post-hoc Dunnett’s multiple comparisons with ipsilateral group ipsi vs. ipsi with 4AP 0 mV to 80 mV p < 0.05, (C) Interaction Group F(1, 1450) = 8.568 p < 0.01 with post-hoc Sidak multiple comparisons contra vs. ipsi −140 mV to 100 mV p > 0.05). (D) Resting membrane potential (RMP) of the patched detached CFP+ cells at 7 days after SNI (contra N = 25 cells, ipsi N = 35 cells, unpaired Student’s t-test p > 0.05). (E) Voltage clamp protocol and representative traces recorded in whole-cell patch clamp from contralateral and ipsilateral cells with and without 4AP application. Western blot for K_v1.1 (F), K_v1.6 (G), with respective ponceau staining for total protein in samples of CFP+ cells sorted by FACS from L3, L4, L5 ipsilateral and contralateral DRG, 7 days after SNI. Quantification of K_v1.1 (H) and K_v1.6 (I) protein level normalised to total protein level (N = 5 samples, paired Student’s t-test p > 0.05).