Journal: The Journal of Physiology
Article Title: Diabetes‐induced microvascular complications at the level of the spinal cord: a contributing factor in diabetic neuropathic pain
doi: 10.1113/JP275067
Figure Lengend Snippet: A , isolated spinal cord endothelial cells demonstrated increased cell death in 50 m m glucose when compared with 50 m m mannitol and 5 m m glucose ( * P < 0.05, *** P < 0.001). VEGF‐A 165 b treatment prevented high glucose‐induced endothelial cell death ( ** P < 0.01). B , IB 4 stained vasculature in the spinal cord of naïve age matched controls was compared with that in ( C ) diabetic + vehicle (arrowheads = vessels smaller than 6 μm) and ( D ) diabetic + VEGFA 165 b. E , there was a significant reduction in total volume of the microvasculature in the spinal cord of the diabetic + vehicle group in addition to ( F ) a reduction in vessel diameter compared with naïve controls ( ** P < 0.01, *** P < 0.001, n = 4 per group). E and F , VEGF‐A 165 b treatment prevented the diabetes‐induced vascular degeneration in the lumbar spinal cord ( ** P < 0.01, n = 4 per group). G , VEGF‐A 165 b treatment also prevents the diabetes‐induced decrease in larger and intermediate microvessels. Scale bar: B–D = 25 μm. [Color figure can be viewed at http://wileyonlinelibrary.com ]
Article Snippet: Primary antibodies, mouse anti‐CD31 (2 μg ml −1 , Abcam; AB24590), rabbit anti‐occludin (5 μg ml −1 , Invitrogen; 71‐1500), mouse anti‐VE cadherin (5 μg ml −1 , BD Biosciences, Franklin Lakes, NJ, USA; 550548), rabbit anti‐VEGFR2 (1:200, 55B11, Cell Signaling), rabbit anti‐Pan VEGF‐A (A20, 1 μg ml −1 , Santa Cruz; sc‐152), mouse anti‐VEGF‐A 165 b (2 μg ml −1 , Abcam; ab‐14994), and rabbit anti‐Actin (1:100, Santa Cruz) antibodies were diluted in blocking solution and incubated overnight at 4°C.
Techniques: Isolation, Staining