Journal: Purinergic Signalling

Article Title: Up-regulation of P2X7 receptors mediating proliferation of Schwann cells after sciatic nerve injury

doi: 10.1007/s11302-015-9445-8

Figure Lengend Snippet: Expression of P2X7R ir in longitudinal sections of normal sciatic nerves. a–c show co-localization of P2X7R ir (red) and S100Aβ ir (green). Note an arrowhead showing a fibre-like structure and an arrow showing a trapezoid structure in a and b; c is the merged image of a and b. Note an arrow indicating a trapezoid structure double labelled by both P2X7R and S100β antibodies (yellow). d–f show co-localization of P2X7R ir (red) and Tuj-1 ir (green). Note an arrow showing a trapezoid structure in d and an arrow showing an axon in e. f is the merged image of d and f; note an arrow indicating a green axon passing through the middle of five trapezoid structures. g–i show co-localization of P2X7R ir (red) and p75NTR ir (green). Note an arrow showing a fibre-like structure in g and h. i is the merged image of g and h. An arrow indicates a double-labelled non-myelinating Schwann cell with P2X7R and p75NTR antibodies (yellow). j–l show co-localization of P2X7R ir (red) and MBP ir (green). Note an arrow showing a fibre-like structure with P2X7R ir, which was not labelled by MBP in l. m–o show co-localization of P2X7R ir (red) and CASPR ir (green). Note that no colocalization of P2X7R ir and CASPR ir was observed. Scale bars in a–c and g–i = 100 μm; scale bars in d–f = 50 μm

Article Snippet: The sections were washed 3–5 min in PBS, and then preincubated in a blocking solution (10 % normal bovine serum, 0.2 % Triton X-100, 0.4 % sodium azide in 0.01 mol/L PBS, pH 7.2) for 30 min followed by incubation with the primary antibodies: P2X7R (1:1000), Alomone, rabbit polyclonal, APR-004; S100β (1:400), Tuj-1 (1:200), p75NTR, PCNA (1:200), MBP (1:200), mouse monoclonal antibodies from Boster, CASPR (1:400) mouse monoclonal antibodies from Santa Cruz, at room temperature overnight.

Techniques: Expressing

Journal: Arthritis Research & Therapy

Article Title: Expression and regulation of neurotrophins in the nondegenerate and degenerate human intervertebral disc

doi: 10.1186/ar2487

Figure Lengend Snippet: Nerve growth factor expression in human intervertebral disc tissue. Nerve growth factor (NGF) protein expression was identified in (a) , (c) the chondrocyte-like cells of human nucleus pulposus and inner annulus fibrosus tissue, and (e) the fibroblast-like cells of the outer annulus fibrosus. (b) , (d) , (f) IgG controls were negative. Scale bar = 50 μm.

Article Snippet: Quantitative real-time PCR was carried out using the predesigned FAM MGB combined primer and the probe sets termed assays on demand for NGF, BDNF and substance P from Applied Biosystems (Warrington, UK): NGF, Hs00171458_m1; BDNF, Hs00542425_s1; substance P, Hs00243225_m1.

Techniques: Expressing

Journal: Arthritis Research & Therapy

Article Title: Expression and regulation of neurotrophins in the nondegenerate and degenerate human intervertebral disc

doi: 10.1186/ar2487

Figure Lengend Snippet: Neurotrophin immunopositivity in the nondegenerate and degenerate intervertebral disc. Histograms illustrating the percentage of (a) positive nerve growth factor (NGF) cells and (b) brain-derived neurotrophic factor (BDNF) immunopositive cells in the nucleus pulposus (NP), inner annulus fibrosus (IAF) and outer annulus fibrosus (OAF) regions of nondegenerate, moderately degenerate and severely degenerate intervertebral disc (IVD) tissue. *Significance between disease states, **significance between regions of the IVD ( P < 0.05). Data presented as the mean ± standard error of the mean.

Article Snippet: Quantitative real-time PCR was carried out using the predesigned FAM MGB combined primer and the probe sets termed assays on demand for NGF, BDNF and substance P from Applied Biosystems (Warrington, UK): NGF, Hs00171458_m1; BDNF, Hs00542425_s1; substance P, Hs00243225_m1.

Techniques: Derivative Assay

Journal: Arthritis Research & Therapy

Article Title: Expression and regulation of neurotrophins in the nondegenerate and degenerate human intervertebral disc

doi: 10.1186/ar2487

Figure Lengend Snippet: Effect of proinflammatory cytokines on neurotrophin and neuropeptide gene expression in nucleus pulposus cells. Histograms illustrating the relative gene expression of (a) nerve growth factor (NGF), (b) brain-derived neurotrophic factor (BDNF) and (c) substance P in nondegenerate and degenerate nucleus pulposus (NP) cells cultured with IL-1β and TNFα normalised to 18s and untreated controls. *Significance between IL-1β treatment and untreated control, **significance between IL-1β and TNFα treatments ( P < 0.05). Data presented as the mean ± standard error of the mean.

Article Snippet: Quantitative real-time PCR was carried out using the predesigned FAM MGB combined primer and the probe sets termed assays on demand for NGF, BDNF and substance P from Applied Biosystems (Warrington, UK): NGF, Hs00171458_m1; BDNF, Hs00542425_s1; substance P, Hs00243225_m1.

Techniques: Gene Expression, Derivative Assay, Cell Culture, Control