Neuropathy is a debilitating condition that affects Diabetic patients. The development of neuropathy is associated with a loss of VEGF-A signaling in the central nervous system and, as a result, dysfunction within the neurovascular interactions. Here, we investigate the loss of spinal cord vasculature and the development of neuropathy in an endothelial cell specific inducible VEGF receptor 2 (VEGFR2) knockdown model in mice. Procedures were carried out in accordance with the UK Home Office Animals Scientific Procedures Act 1986. 24 transgenic mice (C57bl6 30g; both genders) were used. All mice used were vegfr2fl/fl and either Tie2CreERT2 positive (n=12) or Tie2CreERT2 negative (n=12), dosed once daily intraperitoneally (i.p) with 1mg tamoxifen or vehicle (10% ethanol in autoclaved sunflower oil), for 5 consecutive days. Nociceptive behavior was tested using Hargreaves test. After 8 days, hypoxyprobe (60mg/kg) i.p or tritc-dextan intravenously (1mg/20g body weight under sodium pentobarbital 60mg/kg i.p anaesthesia) was injected 30 minutes before euthanasia, then transcardially perfused with 4% paraformaldehyde under terminal anaesthesia (sodium pentobarbital 60mg/kg i.p). Spinal cord tissue was collected and frozen. Cryosections (40μM) were stained using Isolectin B4 (IB4), and Anti-hypoxyprobe. Confocal imaging was performed followed by Imaris 8.1 3D rendering. Values are mean ± S.E.M. One-way ANOVA unless otherwise stated. VegfR2ko (Tie2CreERT2 VEGFR2flfl + tamoxifen; cKO) mice showed a pronounced response to heat as compared to mice treated with vehicle (Tie2CreERT2 VEGFR2flfl+vehicle; Veh) or negative for Tie2CreERT2 (VEGFR2flfl+tamoxifen; WT) (6±2 vs. 8±1s, Veh and 8±1s, WT, p<0.001,n=5). Confocal images of IB4 stained vessels followed by 3D rendering revealed a significant reduction in vessel volume (cKO=400±100 vs Veh=800±100μm3/fieldview, p<0.001 and WT=600±100μm3/fieldview, p<0.05, n=6) and diameter (cKO=2±0.5μm vs. Veh=3.2±0.5, p<0.05, and WT=3.2±0.5, p<0.05, n=6) in the cKO as compared to controls. Confocal images showed an increase in the intensity of hypoxyprobe staining in the cKO versus WT (cKO=1.6±0.2 vs. WT=1.0 ±0.1 x 106 pixels, p<0.05, Unpaired T-test). Mapping of the dorsal horn of the spinal cord showed an increase in hypoxic cells per lamina (lx) in the cKO against WT (l2= cKO=33±6.5 vs. WT=2.5±0.8 p<0.001, l4 = cKO=12±2.2 vs. WT=1±0.4, p<0.05, Two way ANOVA, n=4). A reduction in perfusion was seen in cKO (cKO=3.7±0.5 vs. WT=4.7±0.6 % fluorescence area fraction, p<0.01 Mann-Whitney, n=4) as to WTs. Thus our data suggests a loss of VEGFR2 signaling in the endothelium leads to spinal cord vascular degeneration and hypoxia, which is associated with neuropathic pain development.