Fig. 5

Insulin is essential for cone survival in vitro and delays RD-induced cone loss in vivo. Representative images of peanut agglutinine- (PNA, red) and cone arrestin- (CAR, green) stained retinal explants and quantification of a PNA+ cones and b CAR+ cones after 5 days of culture in high glucose DMEM (25 mM) with or without human insulin and/or insulin receptor inhibitor (HNMPA) or its vehicle (n = 5/group; b one-way Anova *p = 0.0358 and ^$p < 0.0001 versus the high glucose culture; c one-way Anova *p = 0.0012 and ^$p < 0.0001 versus the high glucose culture). c Representative images of peanut agglutinine- (PNA, red), cone arrestin- (CAR, green) and ionized calcium-binding adapter molecule 1- (IBA-1, white) stained retinal flatmounts of C57BL6/J mice 7 days after RD without or with subretinal injection of insulin. Quantification of d subretinal IBA-1+ cells, e PNA+ cones, and f CAR+ cones in retinal flatmounts of C57BL6/J mice without RD and 7 days after RD with or without subretinal injection of insulin (n = 6–12/group; e one-way Anova *p < 0.0001 and ^$p < 0.0001 versus the control group; f one-way Anova/Bonferroni test *p < 0.0001 versus the control group, ^$p = 0.0020 versus the PBS group; g one-way Anova/Bonferroni test *p < 0.0001 versus the control group, ^$p = 0.0002 versus the PBS group). g Representative images of peanut agglutinine- (PNA, red), cone arrestin- (CAR, green), and ionized calcium-binding adapter molecule 1- (IBA-1, white) stained retinal flatmounts of C57BL6/J mice 7 days after RD without or with subretinal injection of IGF-1. Quantification of h subretinal IBA-1+ cells, i PNA+ cones, and j CAR+ cones in retinal flatmounts of C57BL6/J mice without RD and 7 days after RD with or without subretinal injection of IGF-1 (n = 6–12/group; g one-way Anova *p < 0.0001, ^$p < 0.0001 versus the control group; h one-way Anova *p < 0.0001 versus the control group; i one-way Anova *p < 0.0001 versus the control group). PNA, peanut agglutinine; CAR, cone arrestin; IR, insulin receptor; DMSO, dimethyl sulfoxide; IBA-1, ionized calcium-binding adapter molecule 1; PBS, phosphate-buffered saline; RD, retinal detachment; IGF-1, insulin-like growth factor; DMEM, Dulbecco’s modified Eagle’s medium; HNMPA, hydroxy-2-naphthalenylmethylphosphonic acid. Scale bar (a, c, g) = 50 μm. All values are reported as mean ± SEM