(A) CRD at a pressure of 40 mmHg in CVH mice results in activation of DH neurons within the thoracolumbar (T10-L1) spinal cord, as indicated by pERK-IR (yellow arrows). CVH mice pretreated with intracolonic CCDC (100 μM) display more DH neurons in the spinal cord following 40 mmHg CRD. Scale bars: 100 μm. (B) Group data showing that CVH mice pretreated with intracolonic CCDC (100 μM) displayed significantly more pERK-IR DH neurons within the spinal cord following 40 mmHg CRD compared with 40 mmHg CRD alone (****P < 0.0001, dots indicate individual counts in spinal cord sections from N = 6 CVH CRD and N = 6 CVH CCDC+CRD). (C) Representative track paths are shown for individual CVH mice administered either intracolonic vehicle (saline) or an itch cocktail consisting of a combination of CCDC (100 μM), BAM8-22 (20 μM), and CQ (10 μM). (D–F) Intracolonic administration of the itch cocktail to CVH mice significantly reduces (D) their track length covered within the central area of the observation enclosure (**P < 0.01; CVH + vehicle, N = 9; CVH + itch cocktail, N = 7) and (E) reduces their distance from the walls of the enclosure (*P < 0.05; CVH + vehicle, N = 9; CVH + itch cocktail, N = 7) compared with vehicle-treated CVH mice. (F) Intracolonic administration of the itch cocktail to CVH mice also significantly increased grooming behavior compared with CVH vehicle-treated mice (*P < 0.05; CVH + vehicle, N = 9; CVH + itch cocktail, N = 7). Data represent mean ± SEM. Dots represent values from individual mice. P values determined by unpaired t tests (B, D, E, F).