Figure 2. Somatosensory Behaviors of VGLUT3^fl/fl;Lbx1^Cre Mice.

(A) In lsl-tdTom;VGLUT3^Cre mice, tomato⁺ neurons are located in laminae II_i–III and 16% co-localize with PKCγ-IR (arrow).

(B) In VGLUT3^EGFP mice, EGFP is expressed by laminae II_i–III neurons at p10, but not adult.

(C) VGLUT3-IR in VGLUT3^fl/fl;Avil^Cre mice peaks around P10 (middle row).

(D) In Lbx1^Cre mice, only spinal cord neurons express Cre.

(E) EGFP and tomato co-localize at p10 in VGLUT3^EGFP;lsl-tdTom;Lbx1^Cre mice (arrow). A few EGFP⁺ neurons at the lamina IV border do not express tomato⁺ (arrowhead).

(F) Tomato is not present in DRG of adult VGLUT3^EGFP;lsl-tdTom;Lbx1^Cre mice.

(G) VGLUT3-IR in VGLUT3^fl/fl;Lbx1^Cre mice at P10 is present in primary afferents (arrow) and not spinal cord neurons.

(H) Baseline PWT of VGLUT3^fl/fl;Lbx1^Cre mice was similar to controls. After SNI (n=8 both groups), PWTs were significantly elevated compared to controls. Withdrawal responses in the Randall-Selitto assay (n=9 both groups) were also significantly higher than controls. After carrageenan, PWTs of VGLUT3^fl/fl;Lbx1^Cre mice (static mechanical) were elevated compared to controls (n=9 both groups). The mice also responded less frequently than controls to light dynamic mechanical stimulation with a cotton swab (n=11 and n=8 respectively). Latency to respond in the Hargreaves’ test did not differ from controls (n=11 and n=8 respectively). VGLUT3^fl/fl;Lbx1^Cre mice responded less frequently than controls to pinprick of the plantar hindpaw (n=11 and n=8 respectively). Sticky tape and fur clip measures of touch did not differ between VGLUT3^fl/fl;Lbx1^Cre mice and controls (both tests n=8 and n=7 respectively).

All scale bars = 100 μm except inset in E (20 μm). Data are mean ± SEM. *p<0.05, **p ≤ 0.01, ***p ≤ 0.001.