Fig. 1

Location of the initial optic nerve disruption injury in IA-injured animals. a-c. Early axonal and BBB changes as shown with double immunofluorescence for APP (green) and mouse IgG (red). Eyeball is to the left, optic chiasm to the right of panel. Four hours after injury, there are multiple axonal swellings and bulbs (green), evidence of primary TAI at approximately 2/3 of optic nerve from the eyeball. There is also disruption of BBB manifested by IgG leakage (red) in the same area but covering a larger segment of the nerve. There are no APP (+) axonal abnormalities or BBB disruption in the optic nerve of sham mice. Insets show representative lesions in two magnifications at four (b) and 24 (c) hours post-injury. d. Disruption of CTB transport in optic nerve after IA injury: Axonal transport in optic nerve was explored by intravitreally injecting CTB488 1 h after injury. Processed with CLARITY 2 days after injury, CTB-488 (green) transport is interrupted at approximately 2/3 of the optic nerve from the eyeball at exactly the same site as APP (+) axonal abnormalities and BBB alterations in a-c. e-f. Details of axonal abnormalities at the level of initial mechanical nerve disruption. Panels are magnifications of identically labeled regions in b, c, and d. Note classical axonal bulbs 4 h post-injury (e), varicosities and undulations at 24 h (f), and the interrupted axonal transport of CTB in most axons in (g). Scale bars: a-c, 500 μm; d, 550 μm; e-g, 50 μm