Journal: Pain

Article Title: Kidney and brain-expressed protein upregulation in the anterior cingulate cortex mediates chronic post-thoracotomy pain by the phospho-protein kinase Mζ/glutamate receptor 1 signaling pathway and neuroinflammation in male rats

doi: 10.1097/j.pain.0000000000003849

Figure Lengend Snippet: The KIBRA and p -PKMζ are activated in contralateral ACC neurons after CPTP. (A-F) Representative blots and columnar statistical charts show KIBRA, p -PKMζ, and PKMζ levels in contralateral ACC at different time points in Thoracotomy pain (A-C) or Thoracotomy no pain (D-F), * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 vs Sham group; the data were analyzed by 1-way ANOVA followed by Tukey multiple comparisons test, n = 4∼7 rats in each group. (G-H) The photographs of the double immunofluorescence staining show that KIBRA and p -PKMζ are only colocalized with neuron markers (NeuN) but not with astrocyte marker (GFAP) and microglia marker (Iba1) in the Sham and CPTP groups. The white arrowheads indicate the co-immunostaining. Scale bar, 100 μm. ACC, anterior cingulate gyrus; ANOVA, analysis of variance; CPTP, chronic post-thoracotomy pain; GFAP, glial fibrillary acidic protein; Iba1, ionized calcium-binding adapter molecule 1; KIBRA, kidney and brain-expressed protein; NeuN, neuron-specific nuclear protein; PKMζ, protein kinase Mζ; p -PKMζ, phosphorylated protein kinase Mζ.

Article Snippet: Then the samples were dehydrated in 30% sucrose, and cut into slices at 35 μm for immunofluorescence with antibodies against KIBRA (bs-11570R; Bioss), GluR1 (ab174785; Abcam), p -PKMζ (sc174785; Santa Cruz), TNF-α (BS-1857; Bioworld), IL-1β (AF-401-NA; R&D Systems, Minneapolis, MN), neuron-specific nuclear protein (NeuN, MAB377; Millipore, Burlington, VT), glial fibrillary acidic protein (GFAP, 3670s; Cell Signaling Technology, Danvers, MA), ionized calcium-binding adapter molecule 1 (Iba1, ab5076; Abcam), after incubation at 4°C overnight.

Techniques: Double Immunofluorescence Staining, Marker, Immunostaining, Binding Assay

Journal: Pain

Article Title: Kidney and brain-expressed protein upregulation in the anterior cingulate cortex mediates chronic post-thoracotomy pain by the phospho-protein kinase Mζ/glutamate receptor 1 signaling pathway and neuroinflammation in male rats

doi: 10.1097/j.pain.0000000000003849

Figure Lengend Snippet: GluR1 and neuroinflammation in ACC neurons are involved in CPTP. (A-F) Representative blots and columnar statistical charts show GluR1, TNF-α, and IL-1β levels in contralateral ACC at different time points in Thoracotomy pain (A-C) or Thoracotomy no pain group (D-F), * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 vs Sham group, the data were analyzed by 1-way ANOVA followed by Tukey multiple comparisons test, n = 5∼7 rats in each group. (G-I) The photographs of the double immunofluorescence staining show that GluR1, TNF-α, and IL-1β are only colocalized with neuron markers (NeuN) but not with astrocyte marker (GFAP) and microglia marker (Iba1) in the Sham and CPTP groups. The white arrowheads indicate the co-immunostaining: scale bar, 100 μm. ACC, anterior cingulate gyrus; ANOVA, analysis of variance; CPTP, chronic post-thoracotomy pain; GFAP, glial fibrillary acidic protein; GluR1, glutamate receptor 1; Iba1, ionized calcium-binding adapter molecule 1; IL-1β, interleukin-1β; NeuN, neuron-specific nuclear protein; TNF-α, tumor necrosis factor-α.

Article Snippet: Then the samples were dehydrated in 30% sucrose, and cut into slices at 35 μm for immunofluorescence with antibodies against KIBRA (bs-11570R; Bioss), GluR1 (ab174785; Abcam), p -PKMζ (sc174785; Santa Cruz), TNF-α (BS-1857; Bioworld), IL-1β (AF-401-NA; R&D Systems, Minneapolis, MN), neuron-specific nuclear protein (NeuN, MAB377; Millipore, Burlington, VT), glial fibrillary acidic protein (GFAP, 3670s; Cell Signaling Technology, Danvers, MA), ionized calcium-binding adapter molecule 1 (Iba1, ab5076; Abcam), after incubation at 4°C overnight.

Techniques: Double Immunofluorescence Staining, Marker, Immunostaining, Binding Assay