Fig. 3: 3D culture better approximates the presentation of GDNF-RET signaling defects.
From: Engineering kidney developmental trajectory using culture boundary conditions
A Left, Schematic of GDNF-RET signaling and GDNF overactivation via Spry1 knockout. Right, Top: 3D optical projection tomography reconstruction of Spry1−/− kidney UB at E13.5 from ref. 16. B Top, Live fluorescence images of E11.5 Hoxb7-eGFP;Spry1−/− kidney branching in ALI culture from ref. 38. Bottom, Live fluorescence projections of E12 kidneys in 3D culture with exogenous GDNF. C Quantification of 3D culture in (B). (n = 31 and 14 tips from 3 and 4 kidneys for basal and +GDNF conditions, pooled across 1 E12 and 2 E13 litters; unpaired two-tailed t-test, p = 1.33*10−8). D Schematic of RET downregulation effects in culture and in vivo. E Similar images to (B) but for E13 culture in basal media in 3D gel (top), with RET inhibitor in ALI transwell (middle), and with RET inhibitor in 3D gel (bottom). F Quantification of (E). (n = 14, 9, and 9 tubules from 4, 5, and 2 E13 kidneys for basal 3D, Ret-i 3D, and Ret-i ALI cultures, respectively, pooled across 3 litters. One-way ANOVA with Tukey’s test, p = 0.0034 and 8.75*10−5 for basal vs Ret-i media in 3D and 3D vs ALI under RET inhibition.) All plotted as mean ± S.D. **p < 0.01, ***p < 0.001, ****p < 0.0001.
