Journal: Frontiers in Cellular Neuroscience

Article Title: Tear deficiency transforms spatial distribution of corneal calcitonin gene-related peptide-positive nerves in rats

doi: 10.3389/fncel.2025.1619310

Figure Lengend Snippet: Histological alterations in the cornea at different time points after lacrimal gland excision. (A) Light microscopy images of the central cornea after hematoxylin and eosin staining for the sham side at 1 week post-surgery and the excision side at 1, 4 and 8 week post-surgery (Left panel: original magnification, × 40; scale bar, 50 μm. Right panel: magnified images within the dashed line in the left panel.). The arrow indicates the neovascular formations in the stroma. (B) Quantitative analysis of the number of cell nuclei in the stroma. Results are presented as the mean ± SEM ( n = 5 for each group. Statistical significance was assessed using two-way ANOVA, * P < 0.05 and ** P < 0.01, versus the sham side; ## P < 0.01 versus 1-week post-surgery). (C) Light microscopy images of the central cornea after tubulin βIII–immunostaining for the sham side at 1 week post-surgery and the excision side at 1, 4, and 8 weeks post-surgery (Left panel: original magnification, × 40; scale bar, 50 μm. Right panel: magnified images within the dashed line in the left panel.). (D) Quantitative analysis of the tubulin βIII-positive area in the epithelium, representing nerve density. Each column and the vertical bar represent the mean ± SEM ( n = 5 for each group; Two-way ANOVA, * P < 0.05 compared to the sham side, and ## P < 0.01 compared to the 1-week post-surgery).

Article Snippet: Thereafter, the specimens were incubated with anti-tubulin βIII mouse monoclonal (sc-80005; Santa Cruz Biotechnology) and anti-CGRP rabbit polyclonal (Y340; Yanaihara Institute Inc., Shizuoka, Japan) as primary antibodies (both diluted 1:200), for 2 days at 4°C.

Techniques: Light Microscopy, Staining, Immunostaining

Journal: Frontiers in Cellular Neuroscience

Article Title: Tear deficiency transforms spatial distribution of corneal calcitonin gene-related peptide-positive nerves in rats

doi: 10.3389/fncel.2025.1619310

Figure Lengend Snippet: Changes in corneal nerve morphology in whole-mount preparations after lacrimal gland excision. The localization of tubulin βIII–positive nerve fibers (red) and CGRP–positive nerve fibers (green) in the sham side at 1 week post-surgery and the excision side at 1, 4, and 8 weeks post-surgery. (A) Corneal subbasal nerve plexus in the epithelium (original magnification, × 20; scale bar, 50 μm). (B) Corneal nerve innervation in the stroma at a depth of approximately 50–100 μm from the corneal surface (original magnification, × 20; scale bar, 50 μm).

Article Snippet: Thereafter, the specimens were incubated with anti-tubulin βIII mouse monoclonal (sc-80005; Santa Cruz Biotechnology) and anti-CGRP rabbit polyclonal (Y340; Yanaihara Institute Inc., Shizuoka, Japan) as primary antibodies (both diluted 1:200), for 2 days at 4°C.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Tear deficiency transforms spatial distribution of corneal calcitonin gene-related peptide-positive nerves in rats

doi: 10.3389/fncel.2025.1619310

Figure Lengend Snippet: Histological alterations in the cornea at different time points after lacrimal gland excision. (A) Light microscopy images of the central cornea after hematoxylin and eosin staining for the sham side at 1 week post-surgery and the excision side at 1, 4 and 8 week post-surgery (Left panel: original magnification, × 40; scale bar, 50 μm. Right panel: magnified images within the dashed line in the left panel.). The arrow indicates the neovascular formations in the stroma. (B) Quantitative analysis of the number of cell nuclei in the stroma. Results are presented as the mean ± SEM ( n = 5 for each group. Statistical significance was assessed using two-way ANOVA, * P < 0.05 and ** P < 0.01, versus the sham side; ## P < 0.01 versus 1-week post-surgery). (C) Light microscopy images of the central cornea after tubulin βIII–immunostaining for the sham side at 1 week post-surgery and the excision side at 1, 4, and 8 weeks post-surgery (Left panel: original magnification, × 40; scale bar, 50 μm. Right panel: magnified images within the dashed line in the left panel.). (D) Quantitative analysis of the tubulin βIII-positive area in the epithelium, representing nerve density. Each column and the vertical bar represent the mean ± SEM ( n = 5 for each group; Two-way ANOVA, * P < 0.05 compared to the sham side, and ## P < 0.01 compared to the 1-week post-surgery).

Article Snippet: For immunohistochemical staining, the sections were incubated for overnight at 4°C with an anti-tubulin βIII mouse monoclonal antibody (sc-80005; Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:200, an anti-Sema3A mouse monoclonal antibody (sc-74554, diluted 1:1000; Santa Cruz Biotechnology) and an anti-Sema7A mouse monoclonal antibody (sc-374432, diluted 1:400; Santa Cruz Biotechnology).

Techniques: Light Microscopy, Staining, Immunostaining

Journal: Frontiers in Cellular Neuroscience

Article Title: Tear deficiency transforms spatial distribution of corneal calcitonin gene-related peptide-positive nerves in rats

doi: 10.3389/fncel.2025.1619310

Figure Lengend Snippet: Changes in corneal nerve morphology in whole-mount preparations after lacrimal gland excision. The localization of tubulin βIII–positive nerve fibers (red) and CGRP–positive nerve fibers (green) in the sham side at 1 week post-surgery and the excision side at 1, 4, and 8 weeks post-surgery. (A) Corneal subbasal nerve plexus in the epithelium (original magnification, × 20; scale bar, 50 μm). (B) Corneal nerve innervation in the stroma at a depth of approximately 50–100 μm from the corneal surface (original magnification, × 20; scale bar, 50 μm).

Article Snippet: For immunohistochemical staining, the sections were incubated for overnight at 4°C with an anti-tubulin βIII mouse monoclonal antibody (sc-80005; Santa Cruz Biotechnology, Dallas, TX, USA) diluted 1:200, an anti-Sema3A mouse monoclonal antibody (sc-74554, diluted 1:1000; Santa Cruz Biotechnology) and an anti-Sema7A mouse monoclonal antibody (sc-374432, diluted 1:400; Santa Cruz Biotechnology).

Techniques: