Extended Data Fig. 2: Additional behavioral and molecular characterization of ASD mouse models during neonatal ages.
Related to Fig. 2. a, Displacement responses to single air puff stimuli of varying intensities delivered in sequence to the nape of the neck, in P4 animals treated with Nair on the back hairy skin and non-treated animals. At the end of back hairy skin trials, the 1.0 PSI stimulus was delivered to the whisker pad. N = 2 control mice and N = 2 Nair-treated mice. b, Displacement responses to a single 1.0 PSI air puff stimulus before and after topical lidocaine application to the nape of the neck. Each pair of dots represents the same P4 animal pre- and post-application. N = 3 mice. c, Dorsal root ganglia immunohistochemistry showing expression of NeuN and MeCP2 in control mice at P0-P1 and P4, and in Mecp2-/y mice to validate antibody specificity. Scale bar denotes 50 µm. d-e, Spinal cord immunohistochemistry showing expression of VGLUT1-labeled synaptic terminals and GABRB3 in control P0-P1 and adult mice. Scale bars denote 10 µm (d), 5 µm (e, left column), and 2 µm (e, right column). Red arrows denote co-labeled puncta. f, Quantification of VGLUT1 and GABRB3 (top) or VGAT and GABRB3 (bottom) co-labeled puncta, as a fraction of total VGLUT1+ or VGAT+ puncta in a field of view, at P0-P1 or adulthood. Red arrows denote examples of proximity between markers. Each dot represents the average value for one animal, with three spinal cord images taken per animal. N = 3 animals for both conditions. g, Displacement responses to a single presentation of 0.10, 0.25, 0.50, 0.75 and 1.0 PSI stimuli for global mutant animals. Each dot represents the displacement response of a single animal on an air puff trial, and dashes represent means. n = 16 Gabrb3+/+ mice and n = 10 Gabrb3+/- mice; n = 12 Mecp2+/y mice and n = 11 Mecp2-/y mice; n = 17 Nlgn2+/+ mice and n = 15 Nlgn2+/- mice; n = 18 Rorb+/+ mice and n = 17 Rorbh1/+ mice. h, Average displacement responses to 10 presentations of a 1.0 PSI stimulus at 20-30 second interstimulus intervals. n = 17 Nlgn2+/+ mice, n = 15 Nlgn2+/- mice, n = 8 Nlgn2-/- mice, P = 0.6848 for Nlgn2+/+ vs. Nlgn2+/-, P = 0.9749 for Nlgn2+/+ vs. Nlgn2-/-. n = 18 Rorb+/+ mice, n = 17 Rorbh1/+ mice, n = 7 Rorbh1/h1 mice, P = 0.6596 for Rorb+/+ vs. Rorbh1/+, P = 0.2211 for Rorb+/+ vs. Rorbh1/h1, one-way ANOVAs with post hoc Dunnett’s tests. i, Fraction of animals habituating to repeated presentations of 1.0 PSI air puffs. P = 0.7043 for Nlgn2+/+ vs. Nlgn2-/-, P = 0.6495 for Rorb+/+ vs. Rorbh1/h1, one-sided Fisher’s Exact tests. For a, f, and h, data represent means ± s.e.m. Statistical information provided in Supplementary Table 1.
