Figure 7.

CGRP administration does not provoke KCC2 internalization in dorsal horn neurons in either female or male mice. A–D, Representative confocal images of the dorsal horn of spinal cord in female and male mice treated with either 0.1 nmol CGRP or vehicle showing CGRP, IB4, KCC2, and NeuN staining. Scale bars: A–D, 20 μm. a–d, High-magnification images of the areas highlighted in the white rectangle in A–D showing neurons expressing KCC2 (top) together with the average pixel KCC2 intensity plots versus distance to the membrane profile (bottom graphs). Scale bars: a–d, 5 μm. E, Average KCC2 intensity profiles from dorsal horn neurons of female mice treated with vehicle (violet line; n = 8 mice), female mice treated with 0.1 nmol CGRP (purple line; n = 10 mice), male mice treated with vehicle (orange line; n = 7 mice), and male mice treated with 0.1 nmol CGRP (red line; n = 10 mice). Results are presented as the mean per group for visualization purposes since the curves are duplications of the graphs in a–d. F, Membrane KCC2 intensity of the four experimental groups calculated at position zero. No significant differences were observed between vehicle or CGRP-treated female or male mice with Welch's ANOVA; n = 7-10 mice/group.