# Data from: Spatiotemporal changes in Netrin/DSCAM1 signaling dictate axonal projection direction in Drosophila small ventral lateral clock neurons

## Cite this dataset

Liu, Jingjing et al. (2024). Data from: Spatiotemporal changes in Netrin/DSCAM1 signaling dictate axonal projection direction in Drosophila small ventral lateral clock neurons [Dataset]. Dryad. https://doi.org/10.5061/dryad.gf1vhhmxj

## Abstract

Axon projection is a spatial and temporal-specific process in which the growth cone receives environmental signals guiding axons to their final destination. However, the mechanisms underlying changes in axonal projection direction without well-defined landmarks remain elusive. Here, we present evidence showcasing the dynamic nature of axonal projections in *Drosophila*'s small ventral lateral clock neurons (s-LNvs). Our findings reveal that these axons undergo an initial vertical projection in the early larval stage, followed by a subsequent transition to a horizontal projection in the early-to-mid third instar larvae. The vertical projection of s-LNv axons correlates with mushroom body calyx expansion, while the s-LNv-expressed Down syndrome cell adhesion molecule (Dscam1) interacts with Netrins to regulate the horizontal projection. During a specific temporal window, locally newborn dorsal clock neurons (DNs) secrete Netrins, facilitating the transition of axonal projection direction in s-LNvs. Our study establishes a compelling* in vivo *model to probe the mechanisms of axonal projection direction switching in the absence of clear landmarks. These findings underscore the significance of dynamic local microenvironments in the complementary regulation of axonal projection direction transitions.

## README: Data from: Spatiotemporal changes in Netrin/DSCAM1 signaling dictate axonal projection direction in Drosophila small ventral lateral clock neurons

This dataset contains experimental data from a study aimed at investigating the development process of axon projection in s-LNvs. The data includes measurements of vertical axon projection distances and horizontal projection A.U. (arbitrary units) under different experimental conditions. The results of this study are discussed in detail in our relevant publication.

**Description of the data and file structure**

The xlsx data supports Figures 1, 2, 3, 4, 5, 6, Figure 2-Figure supplement 1, Figure 3-figure supplement 1, Figure 3-figure supplement 2, Figure 4-figure supplement 1 Figure 5-figure supplement 1, Figure 6-figure supplement 1.

**Figure1-source data 1 Related to figure1B**: data underlying in the main text. This file contains average vertical projection length at different developmental stages. Figure 1B: Average vertical projection length at different developmental stages

Column B: 8 h

Column C: 24 h

Column D: 48 h

Column E: 72 h

Column F: 96 h

Column G: 120 h

**Figure1-source data 2 Related to Figure 1C**: data underlying in the main text. This file contains average horizontal projection (A.U.) at different developmental stages. Figure 1C: Average horizontal projection at different developmental stages

Column B: 8 h

Column C: 24 h

Column D: 48 h

Column E: 72 h

Column F: 96 h

Column G: 120 h

**Figure2-source data 1 Related to figure2B**: data underlying in the main text. This file contains three tabs. Figure 2B: Development of vertical projection process alongside Mushroom Body growth

Total vertical projection distance

Column B: 24 h

Column C: 48 h

Column D: 72 h

Column E: 96 h

Column F: 120 h

**OK107-GAL4 calyx area**

Column I: 24 h

Column J: 48 h

Column K: 72 h

Column L: 96 h

Column M: 120 h

**201Y-GAL4 calyx area**

Column O: 24 h

Column P: 48 h

Column Q: 72 h

Column R: 96 h

Column S: 120 h

**Figure2-source data 2 Related to figure2E**: data underlying in the main text. This file includes data on the MB under both ablation and non-ablation treatments, with vertical projection distances

Column A: 24 h no ablation

Column B: 24 h ablation

Column C: 48 h no ablation

Column D: 48 h ablation

Column E: 72 h no ablation

Column F: 72 h ablation

**Figure2-supplement figure1 source data 1**: data underlying in the supplement figure. This file includes data on the MB calyx area under both ablation and non-ablation treatments

Column A: 24 h no ablation

Column B: 24 h ablation

Column C: 48 h no ablation

Column D: 48 h ablation

Column E: 72 h no ablation

Column F: 72 h ablation

**Figure3-source data 1 Related to figure3B**: data underlying in the main text. This file includes the vertical projection distances and related statistical methods for both non-knockdown and knockdown Dscam1 treatments.Figure 3B: Critical role of Dscam1 in horizontal projection

Column A: Pdf >GFP

Column B: Pdf >GFP, Dscam1RNAi

**Figure3-source data 2 Related to figure3C**: data underlying in the main text. This file includes the horizontal projection A.U. and related statistical methods for both non-knockdown and knockdown Dscam1 treatments.Figure 3C: Critical role of Dscam1 in horizontal projection

Column A: Pdf >GFP

Column B: Pdf >GFP, Dscam1RNAi

**Figure3-source data 3 Related to figure3E**: data underlying in the main text. This file includes the horizontal projection A.U. and related statistical methods for different Dscam1 mutant fly.

Column A: w1118

Column B: Dscam1 21/+

Column C: Dscam1 1/+

Column D: Dscam1 05518/+

Column E: Dscam1 21/05518*

Column F: Dscam1 21/1

Column G: Dscam1 05518

**Figure3-supplement figure1 source data 1**: data underlying in the supplement figure. This file includes the vertical projection distances and related statistical methods for both non-knockdown and knockdown Dscam1 treatments.

Column A: Clk856-Gal4 >GFP

Column B: Clk856-Gal4 >GFP, Dscam1RNAi

**Figure3-supplement figure1 source data 2**: data underlying in the supplement figure. This file includes the horizontal projection A.U. and related statistical methods for both non-knockdown and knockdown Dscam1 treatments.

Column A: Clk856-Gal4 >GFP

Column B: Clk856-Gal4 >GFP, Dscam1RNAi

**Figure3-supplement figure2 source data 1**: data underlying in the supplement figure. This file includes the horizontal projection A.U.and related statistical methods for Pdf-GAL4 knockdown Dscam1 interaction molecule and cytoskeleton-associated regulatory proteins

Column A: pdf-GAL4/+

Column B: pdf-GAL4 >tsrRNAi

Column C: pdf-GAL4 >chicRNAi

Column D: pdf-GAL4 >SH3PX1RNAi

Column E: pdf-GAL4 >DockRNAi

Column F: pdf-GAL4 > PakRNAi

**Figure4-source data 1 Related to figure4B**: data underlying in the main text. This file includes the horizontal projection A.U. and related statistical methods for netrins mutant

Column A: w1118

Column B: NetAΔ

Column C: NetBΔ

Column D: NetABΔi

**Figure4-source data 2 Related to figure4D**: data underlying in the main text. This file includes the horizontal projection A.U. and related statistical methods for nSyb-GAL4 and repo-GAL4 knockdown Netrins

Column A: UAS-dicer; nSyb-GAL4/w1118

Column B: UAS-dicer; nSyb-GAL4/Netrins RNAi

Column C: UAS-dicer; repo-GAL4/w1118

Column D: UAS-dicer; repo- GAL4/Netrins RNAi

**Figure4-supplement figure1 source data 1**: data underlying in the supplement figure. This file includes the horizontal projection A.U. and related statistical methods for tubulin-GAL4 knockdown Netrins or slit

Column A: tubulin-GAL4/W1118

Column B: tubulin-GAL4/Netrins RNAi

Column C: tubulin-GAL4/slit RNAi

**Figure5-source data 1 Related to figure5B**: data underlying in the main text. This file includes the horizontal projection A.U. and related statistical methods for OK107-GAL4 and per-GAL4,pdf-GAL80;UAS-GFP* knockdown Netrins

Column A: OK107-GAL4/w1118

Column B: OK107-GAL4 /Netrins RNAi

Column C: per-GAL4,pdf-GAL80;UAS-GFP/w1118

Column D: per-GAL4,pdf-GAL80;UAS-GFP/Netrins RNAi

**Figure5-source data 2 Related to figure5D**: data underlying in the main text. This file includes the horizontal projection A.U. and related statistical methods for DN ablation at different developmental stage.

Column A: 72 h no ablation

Column B: 72 h ablation

Column C: 96 h no ablation

Column D: 96 h ablation

**Figure5-source data 3 Related to figure5F**: data underlying in the main text. This file includes the horizontal projection A.U. and related statistical methods for per-GAL4, Pdf-GAL80 overexpress NetB in NetABΔ.

Column A: NetABΔ

Column B: NetABΔ;per-GAL4

Column C: NetABΔ;OE-NetB

**Figure5-supplement figure1 source data 1**: data underlying in the supplement figure. This file includes the horizontal projection A.U. and related statistical methods for both ablated DN or crz+ neuron

Column A: per-GAL4,Pdf-GAL80 No ablation

Column B: per-GAL4,Pdf-GAL80 Ablation

Column C: crz-GAL4 No ablation

Column D: crz-GAL4 Ablation

**Figure6-source data 1 Related to figure6B**: data underlying in the main text. This file includes the cell numbers at different developmental stage.

Column A: 24 h

Column B: 48 h

Column C: 72 h

Column D: 96 h

**Figure6-supplement figure1 source data 1**: data underlying in the supplement figure. This file includes the horizontal projection A.U. and related statistical methods for both R78G02-GAL4 overexpressed netrin.

Column A: R78G02/+

Column B: R78G02 >UAS-NetA

Column C: R78G02 >UAS-NetB

## Funding

Ministry of Science and Technology of the People's Republic of China

National Natural Science Foundation of China

Department of Science and Technology of Guangdong Province