Fig. 4: Rnf146 overexpression in the PFC causes social deficits in mice. | Experimental & Molecular Medicine

Fig. 4: Rnf146 overexpression in the PFC causes social deficits in mice.

From: Dysregulation of the Wnt/β-catenin signaling pathway via Rnf146 upregulation in a VPA-induced mouse model of autism spectrum disorder

Fig. 4

a Diagram for AAV constructs encoding mouse Rnf146 (mRNF146) and eGFP. ITR, inverted terminal repeat sequence; WPRE, Woodchuck hepatitis virus (WHV) posttranscriptional regulatory element. b Schematic diagram of bilateral AAV virus injection in the PFC (left) and an example of a brain slice expressing Rnf146 in the PFC (right). ACC, anterior cingulate cortex; PL, prelimbic cortex; IL, infralimbic cortex. c Left: In the social preference test, Rnf146-overexpressing mice showed a significant preference for a novel conspecific (M) versus an object (O) compared to eGFP-overexpressing control mice (n = 11 mice for eGFP, n = 11 mice for Rnf146, two-way ANOVA: Rnf146, F1, 40 = 0.000, P > 0.9999; target, F1, 40 = 65.87, ****P < 0.0001; interaction, F1, 20 = 6.025, *P = 0.0234; Sidak’s multiple comparisons test: eGFP, mouse versus object t40 = 7.52, ****P < 0.0001; Rnf146, mouse versus object t40 = 3.958, ***P = 0.0006). Right: preference index (PI) of eGFP- or Rnf146-overexpressing mice. Rnf146-overexpressing mice showed PI values comparable to those of control mice (n = 11 mice for eGFP, n = 11 mice for Rnf146, two-tailed unpaired t test; t20 = 1.781, P = 0.0901). d Representative heatmap images from eGFP-overexpressing mice (left) and Rnf146-overexpressing mice (right) of social behavior during the social preference test. e Left: in the social recognition test, Rnf146-overexpressing mice did not show a significant preference for a novel conspecific (N) versus a familiar conspecific (F), while eGFP-overexpressing control mice showed a significant preference for the novel conspecific (n = 11 mice for eGFP, n = 11 mice for Rnf146, two-way ANOVA: Rnf146, F1, 40 = 0, P > 0.9999; target, F1, 40 = 39.94, ****P < 0.0001; interaction, F1, 40 = 15.22, ***P = 0.0004; Sidak’s multiple comparisons test: eGFP, novel versus familiar t40 = 7.227, ****P < 0.0001; Rnf146, novel versus familiar t40 = 1.71, P = 0.1810). Right: PI of eGFP- or Rnf146-overexpressing mice. Rnf146-overexpressing mice showed significantly lower PI values than control mice (n = 11 mice for eGFP, n = 11 mice for Rnf146 group, two-tailed unpaired t test; t20 = 2.759, *P = 0.0121). f Representative heatmap images from eGFP-overexpressing mice (left) and Rnf146-overexpressing mice (right) of social behavior during the social recognition test. g Western blot analysis of the Wnt signaling pathway in the PFC of Rnf146-overexpressing mice. Samples from the PFC of GFP-overexpressing mice were used as controls. h Rnf146-overexpressing mice showed significantly increased Rnf146 levels (normalized to tubulin levels) compared to eGFP-overexpressing mice (n = 7 mice for eGFP, n = 10 mice for Rnf146, two-tailed unpaired t test; t14 = 2.381, *P = 0.0320). i Rnf146-overexpressing mice showed significantly decreased p-β-catenin levels (normalized to β-catenin levels) compared to eGFP-overexpressing mice (n = 7 mice for eGFP, n = 10 mice for Rnf146, two-tailed unpaired t test; t14 = 2.995, **P = 0.0096). j Rnf146-overexpressing mice showed a comparable β-catenin level (normalized to tubulin levels) compared to eGFP-overexpressing mice (n = 7 mice for eGFP, n = 10 mice for Rnf146, two-tailed unpaired t test; t14 = 1.483, P = 0.1601).

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