Fig. 4

Depletion of z-dlx5a in zebrafish embryos causes delayed olfactory differentiation. a. Confocal stacked images of Trcp2::Venus (left, yellow fluorescence) and OMP::CFP (right, cyan fluorescence) zebrafish embryos not injected (left), injected with a control MO (right), or injected with anti-z-dlx5a MO (bottom panels), taken at 72 hpf. Arrows indicate the normal axonal pathway in the control embryos. Asterisks indicate the regions of reduced fluorescence intensity. b. Whole-mount bright field micrographs of injected embryo, showing an overall normal embryonic morphology and growth rate in the injected embryos, compared to the non-injected ones. c. Percentages of embryos showing YFP or CFP fluorescence, over the total of examined ones, comparing not-injected, control injected and MO injected ones. d. RT-PCR analysis on RNA extracted from anti-z-dlx5a MO-treated and control embryos, showing that the MO efficiently generates an inactive splice-variant form of the endogenous mRNA. A scheme of the z-dlx5a gene (Ex1–Ex2–Ex3), the positions of the z-dlx5a MO and the position of the PCR primers (A–D) are reported on the left. e. (on the left) Quantification of the olfactory differentiation phenotype by Real-Time qPCR for differentiation-related mRNAs in a sample of the embryonic heads of MO-injected embryos (grey bars). Embryos injected with control MO were used for comparison (open bars). Normalization is carried out relative to control samples, made = 1. (on the right) Relative abundance of z-hoxA7a and relative abundance of z-hoxA10b mRNAs in whole embryos injected with z-dlx5a MO (grey bars), to monitor developmental progression and exclude a generalized delay.