Figure 3.

CaMKIIβ inhibition in the vCA1 increases stress susceptibility

(A) AAV vectors used for the control construct (AAV-shControl) and CaMKIIβ knockdown (AAV-shCaMKIIβ). ITR, inverted terminal repeats; P_CMV, cytomegalovirus promoter; P_Camk2a, Camk2a promoter; U6, U6 promoter; WPRE, woodchuck hepatitis virus posttranscriptional regulatory element.

(B) Experimental paradigm for behavioral testing. CUMS, chronic ultra-mild stress; SIT, social interaction test; SPT, sucrose preference test; FST, forced swim test.

(C) AAV microinjection into the vCA1. Region-specific expression of GFP in vCA1 is shown. Scale bar, 100 μm.

(D) Specific knockdown of CaMKIIβ but not CaMKIIα by the AAV-shCaMKIIβ.

(E–G) CaMKIIβ knockdown in the vCA1 of B6 mice leads to significantly decreased social interaction time in the SIT (E), decreased sucrose preference in SPT (F), and increased immobility time in FST (G) following CUMS exposure. n = 12–15 mice per group.

(H) AAV vectors engineered to overexpress a constitutively active form of CaMKIIβ (CaMKIIβ-CA) with resistance against shCaMKIIβ (AAV-CaMKIIβ-CA^Res) or a control construct (AAV-mCherry).

(I) Experimental paradigm for behavioral testing.

(J) AAV microinjection into the vCA1. Region-specific expression of GFP and mCherry in the vCA1 is shown. Scale bar, 100 μm.

(K and L) Overexpression of CaMKIIβ-CA^Res together with shCaMKIIβ increases time in the interaction zone in the SIT (K) and tends to increase sucrose preference in the SPT (L) in B6 mice, when compared to the mice overexpressing mCherry together with shCaMKIIβ. n = 14–15 mice per group.

(M) Experimental paradigm for behavioral testing. smSDS, subchronic and mild social defeat stress.

(N) Decreased time in the interaction zone in B6 mice given AAV-shCaMKIIβ following smSDS exposure is prevented by overexpression of CaMKIIβ-CA^Res. n = 15–16 mice per group.

(O) AAV vectors engineered to overexpress a dominant-negative form of CaMKIIβ (AAV-CaMKIIβ-DN) or a control construct (AAV-GFP).

(P) Experimental paradigm for behavioral testing.

(Q) AAV microinjection into the vCA1. Region-specific expression of GFP in vCA1 is shown. Scale bar, 100 μm.

(R–S) Overexpression of CaMKIIβ-DN decreases time in the interaction zone in the SIT (R) and sucrose preference in the SPT (S) in B6 mice following smSDS. n = 13–15 mice per group.

(T) Overexpression of CaMKIIβ-DN decreases Fos levels in the vCA1 of B6 mice following smSDS exposure. n = 6 for all groups.

Two-way ANOVA followed by a Tukey's post hoc test (in E-G, N, R, and S), one-way ANOVA followed by a Tukey's post hoc test (in N), and two-tailed Student's t-test (in K, L, and T) were used for statistical analyses. ∗p < 0.05. Bar graphs show mean ± SEM.

See also Figure S3. Complete statistical summaries are provided in Table S2.