Figure 1. Shank3 functions cell-autonomously in peripheral somatosensory neurons for normal innocuous touch behaviors.

(A) Hairy skin sensitivity was measured using tactile PPI. Percent inhibition of the startle response to a 125 dB noise, when the startle noise is preceded by a light air puff (250 ms ISI). Student’s unpaired t-test or one-way ANOVA with post-hoc Tukey’s test, *, p < 0.05.

(B) Response to a light air puff stimulus alone directed to the back hairy skin. Responses are expressed as percent of startle response to a 125 dB noise. Student’s unpaired t-test or one-way ANOVA with post-hoc Tukey’s test, *, p < 0.05.

(C) Texture discrimination was measured using the textured NORT behavioral assay. A positive value indicates preference for the novel object, compared to the familiar object. Student’s unpaired t-test or one-way ANOVA with post-hoc Tukey’s test, *, p < 0.05.

(D) Open field (OF) test was used as a general measure of exploration and anxiety-like behavior. Shown are representative activity traces in the OF test for mutant mice and control littermates.

(E) Percent time spent in the center of the OF chamber. Student’s unpaired t-test or one-way ANOVA with post-hoc Tukey’s test, *, p < 0.05.

(F) Percent time spent in the open arms of the EPM. Student’s unpaired t-test or one-way ANOVA with post-hoc Tukey’s test, *, p < 0.05.

(G) Preference index for the percentage of time spent investigating a novel mouse, compared to a novel object, in the “Sociability” portion of the 3-chamber social interaction test. Student’s unpaired t-test or one-way ANOVA with post-hoc Tukey’s test, *, p < 0.05; ^#, p < 0.10.

(H) Preference index for the percentage of time spent investigating a novel mouse, compared to a familiar mouse, in the “Social Novelty Preference” portion of the 3-chamber social interaction test. Student’s unpaired t-test or one-way ANOVA with post-hoc Tukey’s test, *, p < 0.05; ^#, p < 0.10.

(I) Voltage step protocol used to activate HCN channels and elicit I_h during whole-cell voltage clamp recordings.

(J-K) Representative electrophysiological traces showing I_h during a hyperpolarizing voltage step protocol in large diameter DRG neurons cultured control and Shank3B^+/− mutant mice, at baseline (K) and with a selective HCN-channel blocker, ZD-7288 (L).

(L) Quantification of I_h density at each voltage step for large diameter neurons cultured from DRGs of control and mutant mice. Two-way ANOVA with post-hoc Sidak’s test, [F (1,1470) =187.7; P < 0.0001] *, p < 0.05.

(M) Representative traces from large diameter DRG neurons cultured from control and Shank3B^+/− mutant mice during whole cell current clamp recordings, in which the minimal amount of current required to elicit an action potential in each neuron (rheobase, R_h), was determined.

(N) Quantification of average R in large diameter DRG neurons cultured from control and Shank3B^+/− mutant mice. Student’s unpaired t-test, *, p < 0.005.