Figure 8.
The relative kinetics of appearance and disappearance of OH-Et or Alexa Fluor 488-annexin V in single cells after optic nerve transection. (A) Flowchart of experiments with HEt. (B) In a total of 124 OH-Et-positive cells studied, 73% were positive for <12 h (black), 27% for 12–24 h and 25% for 24–36 h. (C) Longitudinal retinal CSLO imaging was performed every 12 h after optic nerve transection in eyes following intravitreal injection of HEt. Arrows indicate OH-Et-positive cells that were negative at the next imaging session 12 h later, implying that the duration of superoxide in the cell was <12 h in these cells. The arrowhead indicates an OH-Et-positive cell that was subsequently negative at 24 h, implying that the duration of superoxide in this cell was 12-24 hours. (D) Flowchart of experiments with annexin V. (E) Longitudinal imaging of axotomized retinas in eyes with Alexa Fluor 488-annexin V was performed every 24 h. Of 250 annexin V-positive cells analysed, 43, 41 and 16% remained positive <24, 24–48 and 48–72 h, respectively. Scale bars: 20 µm. ONT = optic nerve transection.