Figure 4.

CCK signaling by Edinger-Westphal neurons is necessary for the switch to preparatory nesting during pregnancy

(A) Sections at the level of the EW nucleus in animals injected in the EW with 3 AAVs expressing saCas9 and one of 3 gRNAs targeting Ucn1, CART, or CCK expression, showing the specific deletion of each gene (an example of the constructs injected is shown for Ucn1). CART and Ucn1 gRNAs lead to almost complete loss of expression, but mice injected with CCK gRNAs still retain some expression. The efficiency of deletions is similar in the triple knockdowns and in the single knockdowns.

(B) Rolling-average nest scores of these animals. Only the triple knockdown has significantly reduced scores.

(C) Proportion of mice transitioning to preparatory nesting in the cohort. CART and CCK knockdowns reduce the proportion of mice consistently building preparatory nests. Scale bars are 150 μm. ∗∗ P < 0.01. All data are presented as mean ± SEM. Dunn’s multiple comparisons test (B) and unpaired Mann-Whitney ranked U test (C). See also Figure S3.