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. 2024 Jan 5;12:RP90133. doi: 10.7554/eLife.90133

Figure 4. Bsh represses Zfh1 to suppress L1/L3 neuronal fate.

(A–A’’) Zfh1 is expressed in all lamina progenitor cells (LPCs) and some lamina neurons at 12 hr after pupa formation (APF). GFP labels all lamina cells. Elav labels lamina neurons. The yellow dash circle outlines LPCs and the white dash circle outlines lamina neurons. Genotype: R27G05-Gal4>UAS-myrGFP. Here and below, scale bar, 10 µm. n≥5 brains. (B–B’’) Zfh1 is expressed in Svp+ L1 and Erm+ L3 neurons at 1–2 days APF; Svp and Erm are never co-expressed. The white dashed circle outlines L1 and L3 neurons and the yellow line indicates the rough boundary between L1 and L3 cell bodies. (C) Schematic of lamina neuron development from 1-day pupae to adult. (D–F) Zfh1-knockdown (KD) in LPCs results in a loss of Svp+ L1 and Erm+ L3 neurons. Quantification: the number of Erm+ or Svp+ cell bodies in a single optical slice. Genotype: R27G05-Gal4>UAS-zfh1-RNAi. (G–I) Bsh-KD in LPCs results in ectopic Zfh1 in L4/L5 layers. White circles label Bsh+ cell bodies in L4 layer in control. L5 layer, white outline. Quantification: the percentage of Bsh+ or Zfh1+ neurons in L5 layer. Genotype: R27G05-Gal4>UAS-bsh-RNAi. (J) Summary. Data are presented as mean ± SEM. Each dot represents each brain. n=6 brains in (F) and n=5 brains in (I). ***p<0.001, unpaired t-test.

Figure 4.

Figure 4—figure supplement 1. Zfh1 expression and epistasis with Bsh.

Figure 4—figure supplement 1.

(A–D) Zfh1-knockdown (KD) in lamina progenitor cells (LPCs) significantly reduces Zfh1 expression in all LPCs and lamina neurons at 12 hr after pupa formation (APF). The white line circle outlines LPCs. The yellow line circle outlines lamina neurons. White dash circle outlines Zfh1+ glia. Scale bar here and below, 10 µm. Genotype: R27G05-Gal4>UAS-Zfh1-RNAi. Quantification: Zfh1 signal of three cell bodies from each brain and normalized Zfh1 signal by setting the highest Zfh1 signal as 100. (E–H) The total number of Ap+ and Pdm3+ cell bodies is decreased when knocking down Zfh1 in LPCs. The number of Bsh+ L4 and L5 neurons is decreased when knocking down Zfh1 in LPCs. Scale bar, 10 µm. (G) The total number of Ap+ and Pdm3+ cell bodies in a single slice. (H) The number of Bsh+ L4 and L5 neurons in a single slice. Genotype: R27G05-Gal4>UAS-Zfh1-RNAi. (I–K) The number of lamina neurons is decreased when knocking down Zfh1 in LPCs. Scale bar, 10 µm. (K) The number of Elav+ cells in a single slice. Genotype: R27G05-Gal4>UAS-Zfh1-RNAi. (L–N) Zfh1 is ectopically expressed in GFP+ neurons. GFP (Bsh-LexA>LexAop-myrGFP) labels LPCs and L5 neurons in control. The white line circle outlines GFP+ neurons. GFP+ LPCs are Tll+ (yellow arrowhead). Scale bar, 10 µm. (N) The percentage of Zfh1+ cells in GFP+ neurons (Elav+) in control (L) and Bsh-KD (M). We divided GFP+ neurons into three domains with equal width from newborn to older neurons. Data are presented as mean ± SEM. Each dot represents a brain. n=5 brains per genotype. **p<0.01, ***p<0.001, unpaired t-test.