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. 2017 May 23;19(8):1532–1544. doi: 10.1016/j.celrep.2017.05.002

Figure 3.

Figure 3

Drd2 Knockdown in the Auditory Thalamus Rescues Deficits in Associative Fear Memory and Thalamo-LA Synaptic Transmission in 22q11DS Mice

(A) In vivo infection of MGm neurons with lentiviral vector LV-control shRNA-GFP (control shRNA) or LV-Drd2 shRNA-GFP (Drd2 shRNA).

(B) Representative images of a coronal brain section from a mouse infected with control shRNA showing GFP+ neurons in the MGm under low (left) and high (right) magnifications. Sections were counterstained with DAPI (nuclei).

(C) Relative Drd2 mRNA levels in WT and Df(16)1/+ mice injected with control or Drd2 shRNA (WT injected with control shRNA, six mice; Df(16)1/+ injected with control shRNA, six mice; WT injected with Drd2 shRNA, five mice; Df(16)1/+ injected with Drd2 shRNA, five mice). Mann-Whitney rank-sum test: for WT injected with control shRNA versus Df(16)1/+ injected with control shRNA, U = 5, ∗p = 0.041; for Df(16)1/+ injected with control shRNA versus Df(16)1/+ injected with Drd2 shRNA, U = 4, ∗p = 0.026.

(D) Freezing behavior before (pre-CS) and during (post-CS) presentation of the CS in WT and Df(16)1/+ mice injected with control or Drd2 shRNAs (WT injected with control shRNA, 11 mice; Df(16)1/+ injected with control shRNA, 10 mice; WT injected with Drd2 shRNA, 14 mice; Df(16)1/+ injected with Drd2 shRNA, 14 mice). Pre-CS: Kruskal-Wallis one-way ANOVA on ranks, H3 = 7.677, p = 0.53. Post-CS: one-way ANOVA, F3 = 8.786, ∗p < 0.001. Post hoc: Df(16)1/+ injected with control shRNA versus Df(16)1/+ injected with Drd2 shRNA, ∗p = 0.033; WT injected with control shRNA versus Df(16)1/+ injected with control shRNA, ∗∗p = 0.000013.

(E) Active avoidance performance in WT and Df(16)1/+ mice injected with control or Drd2 shRNAs (WT injected with control shRNA, 10 mice; Df(16)1/+ injected with control shRNA, 10 mice; WT injected with Drd2 shRNA, 14 mice; Df(16)1/+ injected with Drd2 shRNA, 11 mice). Two-way repeated-measures ANOVA: F3,3 = 7.392, ∗p < 0.001. Post hoc at day 4: Df(16)1/+ injected with control shRNA versus Df(16)1/+ injected with Drd2 shRNA, ∗p = 0.006; WT injected with control shRNA versus Df(16)1/+ injected with Drd2 shRNA, p = 0.338.

(F) Total number of crossings between compartments in WT and Df(16)1/+ mice injected with control or Drd2 shRNAs. The same number of mice as used in (E). One-way ANOVA: F3 = 1.626, p = 0.198.

(G) EPSC peak amplitude, as a function of stimulation intensity at thalamo-LA projections of the following groups are shown: WT injected with control shRNA (11 neurons, three mice), Df(16)1/+ injected with control shRNA (13 neurons, three mice), WT injected with Drd2 shRNA (13 neurons, three mice), and Df(16)1/+ injected with Drd2 shRNA (9 neurons, three mice). Two-way repeated-measures ANOVA: F3,14 = 8.673, p < 0.001. Post hoc: ∗p < 0.01.

(H) PPR at thalamo-LA projections of the following groups are shown: WT injected with control shRNA (12 neurons, three mice), Df(16)1/+ injected with control shRNA (15 neurons, three mice), WT injected with Drd2 shRNA (15 neurons, three mice), and Df(16)1/+ injected with Drd2 shRNA (10 neurons, three mice). Two-way repeated-measures ANOVA: F3,4 = 20.75, p < 0.001. Post hoc: ∗p < 0.001. Insets show representative pairs of thalamo-LA EPSCs. Scale bar, 100 pA, 50 ms. ns, not significant. Data are represented as mean ± SEM.

See also Figure S7.