Dual control of dopamine in Drosophila myeloid-like progenitor cell proliferation and regulation of lymph gland growth (2022)
Figure 2

Figure 2: The medullary zone progenitors synthesize dopamine to control lymph gland size and differentiation.

(A) Schematic representation of the dopamine synthesis pathway.

(B-D') A temporal expression profile of TH at 48, 72 and 96h AEL lymph glands in control background (domeMESO-Gal4, UAS-GFP/+)

(E-G') A temporal expression profile of Ddc at 48, 72 and 96h AEL lymph glands in control background (domeMESO-Gal4, UAS-GFP/+).

(H) Quantification of TH levels (domeMESO-Gal4, UAS-GFP/+, n=18 at 48h, n= 15, at 72h and n= 23 at 96h AEL).

(I) Quantification of Ddc levels (domeMESO-Gal4, UAS-GFP/+, n= 17 at 48h, n= 33 at 72h and n= 38 at 96h AEL).

(J-L) Assessment of dopamine levels upon the knockdown of TH and Ddc in the progenitors using domeMESO-Gal4, UAS-GFP.

(M) The mean intensity quantification of dopamine levels upon TH and Ddc loss of function in the progenitors (domeMESO-Gal4, UAS-GFP/+, n=40, domeMESO-Gal4, UAS-GFP; THRNAi, n= 17, domeMESO-Gal4, UAS-GFP; DdcRNAi, n= 34).

(N-P) The loss of progenitor TH and Ddc function using domeMESO-Gal4, UAS-GFP analysed at wandering 3rd instar, 120h AEL.

(Q) Quantifications of lymph gland size upon loss of TH (domeMESO-Gal4, UAS-GFP; THRNAi, n= 34) and Ddc (domeMESO-Gal4, UAS-GFP; DdcRNAi, n= 56) when compared to stage matched control (domeMESO-Gal4, UAS-GFP/+, n=90).

(R) Quantifications of the proportions of %Dome (green bars), %negative (blue bar), %overlap (Dome and P1, yellow bars) and %P1 (red bars). For Dome positive area (domeMESO-Gal4, UAS-GFP/+, n=27, domeMESO-Gal4, UAS-GFP; THRNAi, n= 15 and domeMESO-Gal4, UAS-GFP; DdcRNAi, n= 18), negative and overlap areas (domeMESO-Gal4, UAS-GFP; THRNAi, n= 15, domeMESO-Gal4, UAS-GFP; DdcRNAi, n= 18) and P1 positive area (domeMESO-Gal4, UAS-GFP/+, n=27, domeMESO-Gal4, UAS-GFP; THRNAi, n= 15 and domeMESO-Gal4, UAS-GFP; DdcRNAi, n= 18).

(S) Quantification of the number of PPO positive crystal cells upon dopamine synthesis perturbation (domeMESO-Gal4, UAS-GFP/+, n=29, domeMESO-Gal4, UAS-GFP; THRNAi, n= 21, domeMESO-Gal4, UAS-GFP; DdcRNAi, n= 23).

Data information: DNA is stained with Hoechst in blue, iDopamine in red, Tyrosine hydroxylase (TH) and Dopa decarboxylase (Ddc) in red, domeMESO (representative of progenitors) expression in green, P1 (representative of mature plasmatocytes) in red. All image panels show a 20µm scale bar. AEL indicates After Egg Laying. The quantifications in Q and S represent mean with standard deviation, R represents mean and in H, I and M represents the median with whiskers indicating maximum and minimum values, box indicates the lower and upper quartiles. The statistical analysis applied in H, I, M, Q and S is One-way ANOVA with Tukey's multiple comparisons test while that applied in R is Two-way Anova with Tukey's multiple comparisons test. 'n' represents the number of lymph gland lobes analyzed, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. For better representation, the primary lobe of the lymph gland has been represented and outlined in white dashed lines while the MZ is outlined in yellow dashed line.