Figure 1.

Experimental design for water avoidance stress. (a) Water avoidance stress (WAS, one hour per day for four days) was applied during the remission phase of PLP-induced relapsing-remitting EAE mice and the clinical scoring was assessed during 60 days. (b) Photography of the experimental system. Mice were placed for 1 hour on a platform positioned at the center of a plastic container filled with water at room temperature up to 1 cm below the platform level. Mice were randomly assigned to the two experimental groups. Non-WAS mice were handled identically, but placed in a standard cage. (c) Measure of fecal pellet outputs in the two experimental groups as an index of stress-induced colonic motility (n = 26 and n = 28 for EAE and EAE+WAS groups respectively). ***P<0.001; ****P<0.0001, Mann Whitney’s U-test.