Fig. 5

Fig. 5

Insulin knockdown impairs α cell development and destabilizes α cell fate. (A,B) Confocal projections of 100 hpf Tg(insa:CFP-NTR) control (A) and insaMO-injected (B) islets stained for Glucagon (red) and CFP (β cells, green). α cell count is decreased and α/beta; cell architecture is inverted. (C) Quantification of Glucagon-positive cells in 54 hpf, 80 hpf and 100 hpf control (gray) and insaMO-injected (red) islets (n ≥ 5 for all time points except 54 hpf control, n=3). (D–E′) Confocal projections (D,E) and confocal planes (D′,E′) of 2 months post fertilization (mpf) control (D,D′) and insaMO-injected (E,E′) islets stained for Glucagon (red), Insulin (green) and DNA (blue). Reduction of α cell mass was sustained into adult stages. (F,G) Confocal planes of 100 hpf control (F) and insaMO-injected (G) islets immunostained for Glucagon (white) and Pdx1 (red). Double positive Glucagon+ Pdx1+ cells (arrows) were increased with insaMO-injection. (H) Quantification of double positive Glucagon+ Pdx1+ cells in 100 hpf control (n=6) and insaMO-injected (n=10) islets. Two way ANOVA was used in C and Student′s t-test was used in H to determine significance.