Fig. 1: DCNs transcriptome analysis with subsequent RNAi screen of 74 upregulated genes. | Nature Communications

Fig. 1: DCNs transcriptome analysis with subsequent RNAi screen of 74 upregulated genes.

From: Temporal transcriptional regulation of mitochondrial morphology primes activity-dependent circuit connectivity

Fig. 1

A, A’ Multi Color Flip-Out (MCFO) staining of single DCN cell innervating Medulla (M-DCN) and Lobula (L-DCN). B Laser Micro Dissection of DCN clusters from brain cryosections of ato-Gal414A;UAS-CD8::GFP,UAS-RedStinger adult flies. 1, 1’- RedStinger fluorescent DCNs before (1, UV light) and after (1’, UV light, 1”, transmitted light) dissection. 2, 2’ - sample with dissected DCNs clusters in the transmitted (2) and UV (2’) light. Total RNA was extracted using Ambion RNAqueous Micro Kit and libraries were generated using SMART-seq2 Kit and Nextera Tagmentation Kit. NGS NextSeq 500 High Output Kit (400 million reads of apr. 25 million reads/sample coverage was used for sequencing). Created in BioRender. Hassan, B. (2025) https://BioRender.com/fma66dn. C Example of the marker gene atonal high-level expression in DCN samples compared to random central brain regions used as a control, with the equal expression of housekeeping genes. D Heatmap of top-20 genes expressed in DCNs. E–H RNAi-screen of 74 DCN-enriched genes revealed 10 gene candidates causing significant phenotypical changes upon DCN-specific downregulation during development. Lo lobula, OC optic chiasm, Me medulla, DCNs Dorsal Cluster Neurons, VCNs Ventral Cluster Neurons. Scale bar – 20 µm. Genes expressed in DCNs and list of genes for the RNAi-screen are listed in Supplementary Fig. S1. Results of RNAi screen of 74 genes in Supplementary Fig. S1.

Back to article page