Fig. 2

Lef1 activates expression of zebrafish hypothalamic genes associated with anxiety.

(A) Volcano plot of zebrafish RNA sequencing (RNA-seq) shows differentially expressed genes in the 3 days post-fertilization (dpf) hypothalamus of lef1 mutants compared to control. Only genes with adjusted P value (AdjP) <0.1 (green line) are shown. Genes with an absolute value of log₂ ratio >1 (blue lines) are shown in red; others are shown in black. Node size represents the averaged fragments per kilobase of transcript per million mapped reads of a gene in the control. (B) Ingenuity Pathway Analysis (IPA) for zebrafish hypothalamic Lef1-dependent genes revealed 20 genes associated with anxiety and depressive disorder, listed in the table. (C and D) Representative images of whole mount in situ hybridization on 3 dpf control and lef1 mutant embryos for known Wnt targets (C) and genes associated with anxiety and depressive disorder (D). Red and yellow arrows indicate expression in caudal and rostral hypothalamus, respectively. Lateral (axin2, dkk1b, lef1, notum1a, crhbp, and grin2cb) or ventral (other genes) views were selected for optimal expression visualization. Scale bar: 100 μm.