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Abstract
Purpose
To evaluate the expression patterns of F4/80 and nestin in the ciliary body and the optic nerve following N-meth-yl-N-nitrosourea (NMU)-induced retinal degeneration in adult mice.
Methods
After intraperitoneal injection of MNU (60 mg/kg) in adult mice, the eyes were enucleated at 2, 4, 7 and 30 days. Hematoxylin and eosin (H&E) stain, terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) stain and immunohistochemical stains of F/80 and nestin were performed.
Results
After MNU treatment, the photoreceptors were destroyed by cell apoptosis. According to immunohistochemistry, F4/80 and nestin were not co-expressed in the control group, but F4/80 was expressed within the ciliary body and optic nerve in the MNU-treated group; the expression of nestin also increased. In the outer nuclear layer, F4/80 and nestin co-expressing cells were observed.
References
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Figure 1.
H&E staining results of the retina. (A) Normal. (B) 2 days after MNU treatment, the retinal thickness is slightly decreased and the arrangement of PRL and ONL is distorted. (C) 7 days after MNU treatment, the depletion of PRL and RPE are noticed and the ONL is reduced to a few layers of cells. (D) 30 days after MNU treatment, the ONL completely disappeared (RPE = retinal pigment epithelium; PRL = photoreceptor layer; ONL = outer nuclear layer; OPL = outer plexiform layer; INL = inner nuclear layer; IPL = inner plexiform layer; GCL = ganglion cell layer).
Figure 2.
TUNEL staining results of the retina. (A) No TUNEL+ cells are observed in the control group. (B) Large number of TUNEL+ cells are shown in the ONL at 2 days after MNU treatment. (C) Seven days after MNU treatment, small num-ber of TUNEL+ cells are remained in the ONL and GCL. (D) Thirty days after MNU treatment, no TUNEL+ cells are observed as the ONL disappeared (RPE = retinal pigment epithelium; PRL = photoreceptor layer; ONL = outer nuclear layer; OPL = outer plexiform layer; INL = inner nuclear layer; IPL = inner plexiform layer; GCL = ganglion cell layer).
Figure 3.
Immunochemistry examination of the optic nerve head. Immunofluorescent labeling with F4/80 (red) and nestin (green). In the control optic nerve head, there are some of F4/80+ cells (A: arrowhead) and nestin+ cells (B: asterisk), but no cells co-ex-pressed F4/80 and nestin (C: merged image of A and B). Two days after MNU treatment, some of F4/80+ cells are also positive for nestin (D-F: arrow). Four days after MNU treatment, large number of F4/80 and nestin co-expressing cells are observed (G-I: arrow). Seven days after MNU treatment, these cells are decreased (J-L). Scale bar: 100 μ m.
Figure 4.
Immunochemistry examination of the ciliary body. Immunofluorescent labeling with F4/80 (red) and nestin (green). In the control ciliary body, F4/80 immunoactivity is seen in the ciliary epithelium, but nestin is not expressed (A-C: arrow). The stroma of ciliary body is non-specifically stained by nestin, but F4/80 is not expressed (B, E, H: asterisk). In the choroid, F4/80 and nestin co-expressing cells are observed (A-C: arrowhead). Two days after MNU treatment, F4/80 immunoactivity of ciliary epithelium is more increased (D: arrow) and there are some of cells which co-expressed F4/80 and nestin (D-F: arrowhead). Four days after MNU treatment, F4/80+ cells are seen in the ciliary epithelium (G: arrow), and F4/80 and nestin co-expressing cells are seen in the fibrovascular tissue of ciliray body (G-L: arrowhead). (CB: ciliary body). Scale bar: 100 μ m.
Figure 5.
Immunochemistry examination of the retina. Immunofluorescent labeling with F4/80 (red) and nestin (green). In the con-trol retina, there are no F4/80+ cells (A), but nestin is found in retinal blood vessels (B: arrowheads) and muller cells (B: asterisk). Two days after MNU treatment, F4/80 and nestin are co-expressed between INL and ONL (D-F: arrow). Four days after MNU treatment, F4/80 is also expressed in the ONL (G-I: arrow). Nestin immunoactivity of the muller cells was increased (E, H) (PRL = photoreceptor layer; ONL = outer nuclear layer; OPL = outer plexiform layer; INL = inner nuclear layer; IPL = inner plexi-form layer; GCL = ganglion cell layer). Scale bar: 50 μ m.
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