|
Fig. 7 Prucalopride promotes enteric neurogenesis in normal development and injury. (A) After photoconversion of all enteric neurons at 4.5 dpf, cohorts of Phox2b-kaede fish were exposed to 10 µM prucalopride (n=5), 100 µM prucalopride (n=6) or DMSO (n=4) for 12 h and then live imaged at 5 dpf. The mean number of de novo hindgut neurons was significantly higher in fish treated with prucalopride (25, 27.7 and 14, respectively; P=0.019 and P=0.0036, respectively). (B) Under a similar design as A, fish were instead exposed to 100 µM GR 113808 (n=6) versus DMSO (n=6) for 12 h and then re-imaged at 5 dpf. The mean number of de novo hindgut neurons was significantly lower in fish treated with GR 113808 than in controls (8.67 versus 14.33; P=0.0086). (C) At 4.5 dpf, Phox2b-kaede fish underwent laser ablation of ten distal hindgut enteric neurons and then photoconversion of all enteric neurons. Cohorts were exposed to 10 µM prucalopride (n=6) or DMSO (n=5) for 12 h and then live imaged at 5 dpf. There was no difference in de novo distal hindgut neurons between these two groups. (D) Under a similar experimental design as C, fish were instead exposed to 10 µM prucalopride (n=3) or DMSO (n=3) at 3.5 dpf for 12 h, and then underwent cell ablation and photoconversion at 4.5 dpf. At 5 dpf, live imaging revealed significantly more distal hindgut neurons in fish pre-treated with prucalopride (22 versus 13.3; P=0.031). (E) Fish were exposed to 100 µM GR 113808 (n=6) or DMSO (n=6) at 3.5 dpf for 12 h, and then underwent cell ablation and photoconversion at 4.5 dpf. At 5 dpf, live imaging revealed significantly fewer distal hindgut neurons in fish pre-treated with GR 113808 than in controls (5.67 versus 12.50; P<0.0001). Data are mean±s.e.m. *P<0.05, **P<0.01,****P<0.0001. ns, not significant.