Immunofluorescent staining for VGLUT1 (A–C) and VGLUT2 (D–I) in the rat dental pulp in the control (A, D, E), CFA 1-day (B, F), and CFA 3-day (C, G–I) groups. A–C:

<p>Expression of VGLUT1 in pulpal axons in control (A) and CFA 1-day (B) and CFA 3-day (C) groups. VGLUT1 is expressed in many axons in the peripheral portion of the coronal pulp. <b>D–I:</b> Expression of VGLUT2 in pulpal axons in control (D, E), CFA 1-day (F), and CFA 3-day (G–I) groups. In the control group, VGLUT2 is expressed in a small number of axons in the peripheral portion of coronal pulp (D) and few axons in the radicular pulp (E). However, it is expressed in a large number of axons in the peripheral portion (F, G), the core of the coronal pulp (H), and in radicular pulp (I) in the CFA 1-day and CFA 3-day groups. Scale bar = 20 µm.</p

Density of VGLUT1+ and VGLUT2+ axons (A, immunofluorescence assay) and protein levels of VGLUT1 and VGLUT2 (B, C, Western blot assay) in the control, CFA 1-day and 3-day pulps.

<p><b>A</b>: The density (area fraction) of VGLUT1+ pulpal axons is not significantly different between CFA 1-day or CFA 3-day groups and control, whereas the density of VGLUT2+ axons is significantly higher in the CFA 1-day, CFA 3-day groups than control. N = 3 animals in each group. *p<0.01. <b>B</b>: Representative images of the Western blot assay. <b>C</b>: Quantitative analysis of VGLUT1 and VGLUT2 protein in the dental pulp. Protein levels of VGLUT1 and VGLUT2 in the pulp are significantly higher in the CFA 1-day, CFA 3-day groups than control. This difference is bigger for VGLUT2 (3.9 and 4.3 fold higher in the CFA 1-day and CFA 3-day groups than control) than for VGLUT1 (2.9 and 3.1 fold higher in the CFA 1-day and CFA 3-day groups than control). N = 5 animals in each group. *p<0.01.</p

Immunofluorescent staining for VGLUT1 and VGLUT2 (A), density of VGLUT1+ and VGLUT2+ somata (B, immunofluorescence assay), and protein levels of VGLUT1 and VGLUT2 (C, D, Western blot assay) in the trigeminal ganglion.

<p><b>A:</b> Immunofluorescent staining for VGLUT1 (a, b) and VGLUT2 (c, d) in the rat trigeminal ganglion in the control (a, c) and CFA 1-day (b, d) groups. VGLUT1 is expressed predominantly in medium- and large-sized somata (a, b), whereas VGLUT2 is expressed predominantly in small- and medium-sized somata (c, d). The number of VGLUT1+ somata of all somata is not different between control (a) and CFA 1-day group (b), whereas that of VGLUT2+ soma is significantly higher in the CFA 1-day (d) than in the control (c) groups. Scale bar = 50 µm. <b>B:</b> The density of VGLUT2+ somata (fraction of all somata) is significantly higher for the CFA 1-day, CFA 3-day groups than control, whereas the density of VGLUT1+ soma is not significantly different between CFA 1-day or CFA 3-day groups and control. N = 3 animals in each group. *p<0.01. <b>C:</b> Representative images of the Western blot assay for VGLUT1 and VGLUT2 in the rat trigeminal ganglion. <b>D:</b> Quantitative analysis of VGLUT1 and VGLUT2 protein in the trigeminal ganglion. The VGLUT2 protein levels are significantly higher in the CFA 1-day and CFA 3-day groups than for the control, whereas the VGLUT1 protein levels are not different between CFA 1-day or CFA 3-day groups and the control group. N = 5 animals in each group. *p<0.01.</p

Double immunofluorescent staining for VGLUT1 and CD64 (A) and for VGLUT2 and CD64 (B) in the rat dental pulp in CFA 1-day group.

<p>VGLUT1 and VGLUT2 are expressed in many CD64+ cells (arrowhead) as well as in axons (arrow) in the inflamed dental pulp. Scale bar = 20 µm.</p