Fig 1.
Flowchart of the experimental design.
ONC, optic nerve crush; BMD, brimonidine; QD, once daily; BID, twice daily.
Fig 2.
Sampling method for cell counting in retinal whole-mounts.
DAPI-, NeuN-, and GFAP-positive cells were counted in 15 randomly chosen sampling boxes (0.04 mm2): the central (n = 5, red squares), mid-periphery (n = 5, orange squares), and peripheral (n = 5, yellow squares) regions of the retina. DAPI, 4′,6-diamidino-2-phenylindole; NeuN, neuronal nuclei; GFAP, glial fibrillary acidic protein. Scale bar 1000 μm.
Fig 3.
Photomicrograph illustrating immunofluorescence staining of mouse retinal whole-mount in experimental ONC groups, representing retinal cell densities in the experimental ONC and control groups.
A, B, C, D: Grey scale DAPI nuclear marker representing all cells of retinal ganglion cell layer; E, F, G, H ‐ NeuN-stained retinal ganglion cells and amacrine cells in green; I, J, K, L–retinal astrocytes (in red) labeled with antibodies against GFAP. A, E, I: retinal whole-mount explan from control group (without ONC); B, F, J: retinal whole mount explant from ONC/saline drop + IP group; C, G, K: retinal whole mount in ONC/BMD drop group; D, H, L: retinal whole mount in ONC/BMD drop + IP group. ONC, optic nerve crush; BMD, brimonidine; IP, intraperitoneal; DAPI, 4′,6-diamidino-2-phenylindole; NeuN, neuronal nuclei; GFAP, glial fibrillary acidic protein. Scale bar– 50 μm.
Fig 4.
NeuN-stained cell counts in the retina after ONC.
**** p<0.0001. ONC, optic nerve crush; BMD, brimonidine; IP, intraperitoneal; NeuN, neuronal nuclei.
Table 1.
NeuN-positive cell count average (cells/mm2 ± SD) in the total and different regions of murine retina.
Fig 5.
NeuN-stained cell count in the peripheral region of the mouse retina after ONC.
ONC, optic nerve crush; BMD, brimonidine; IP, intraperitoneal; NeuN, neuronal nuclei. * p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001.
Table 2.
Cell count dynamics between retinal regions (Δ cell change ± SD).