Fig. 8: Retinoic acid negatively regulates ret.
From: Axon targeting of transcriptionally distinct pioneer neurons is regulated by retinoic acid signaling
A−C Representative single confocal Z-slices of pLL ganglia expressing EGFP (A), caRAR (B), or dnRAR (C) in individual cells (dashed outlines). Expression of ret (yellow) and hoxb5a (magenta) was assessed by FISH. D, E Quantified expression levels of hoxb5a and ret after injection of RAR constructs. We analyzed the following numbers of embryos from two independent experiments: EGFP control: 21; caRAR: 15, dnRAR: 12. Each plot shows minimum, Q1, median, Q3, and maximum. Nonparametric Kruskal–Wallis test values are shown on plots: *p < 0.05; ****p < 0.0001. Exact p-values are as follows: hoxb5a, GFP vs caRAR < 0.0001; GFP vs dnRAR = 0.0340; caRAR vs dnRAR < 0.0001; ret, GFP vs caRAR = 0.0135; GFP vs dnRAR = 0.1024; caRAR vs dnRAR < 0.0001. F Schematic of mosaic labeling using coinjection of two constructs: caRAR-2A-EGFP and mCherry (control) or caRAR-2A-EGFP and ret-mCherry driven by neurod1 promoter. Following injection, embryos were screened at 72 hpf for the position of colabeled axon terminals. Created in BioRender. Lab, N. (2025) https://BioRender.com/vij5z6b. G, H Confocal image of the neuron cell body (G) and its axon terminal (L1) labeled by caRAR and mCherry (H,H’). I, J Confocal image of the neuron cell body (I) and its axon terminal (terminal NMs) labeled by caRAR and ret-mCherry (J,J’). K, L Frequency of NM targeted by labeled neurons coexpressing either caRAR-2A-EGFP+mCherry or caRAR-2A-EGFP + ret-mCherry: p = 0.0002, Chi-square test. The following number of cells were examined in 2 independent experiments (1 cell/animal): caRAR-2A-EGFP + mCherry – 10; caRAR – 10, and caRAR-2A-EGFP + ret-mCherry – 18. All images are lateral views with anterior to the left. Scale bars = 20 μm. All images scaled the same.
