Journal: Journal of Neuroinflammation

Article Title: Activation of GPR40 produces mechanical antiallodynia via the spinal glial interleukin-10/β-endorphin pathway

doi: 10.1186/s12974-019-1457-9

Figure Lengend Snippet: Expression of GPR40 in microglia ( a – c ), astrocytes ( d – f ), and neurons ( g – i ) in the spinal dorsal horn of neuropathic rats. Frozen sections of the spinal lumbar enlargements were obtained approximately 2 weeks after spinal nerve ligation. Immunofluorescence was double stained with GPR40/Iba-1, GPR40/GFAP, and GPR40/NeuN, and photomicrographs were taken from the spinal cord section ( a , d , g ; 500 μm) and amplified dorsal horn laminae I–V ( b , c , e , f , h , i ; 50 μm). Arrows indicate double immunostaining of GPR40 with each cellular biomarker. Double immunolabeled surface areas of GPR40/Iba-1 ( j ), GPR40/GFAP ( k ), and GPR40/NeuN ( l ) from the spinal dorsal horn laminae I–V indicated in white lines were quantified using the ImageJ program. Data are presented as means ± SEM ( N = 5–6 per group). * P < 0.05 vs. contralateral side; analyzed by unpaired and two-tailed Student’s t test

Article Snippet: Iba-1 , Merck Millipore, Darmstadt, Germany , 1:300 , 1:100.

Techniques: Expressing, Ligation, Immunofluorescence, Staining, Amplification, Double Immunostaining, Biomarker Assay, Immunolabeling, Two Tailed Test

Journal: Journal of Neuroinflammation

Article Title: Activation of GPR40 produces mechanical antiallodynia via the spinal glial interleukin-10/β-endorphin pathway

doi: 10.1186/s12974-019-1457-9

Figure Lengend Snippet: Effect of intrathecal GW9508 on spinal IL-10 expression on spinal microglia ( a – f ), astrocytes ( g – l ), and neurons ( m – r ) in neuropathic rats induced by spinal nerve ligation. Frozen sections of the spinal lumbar enlargements were obtained 1 h after intrathecal saline (10 μl) or GW9508 (30 μg) treatment. Immunofluorescence was double stained with IL-10/Iba-1, IL-10/GFAP, and IL-10/NeuN, and photomicrographs were taken from the entire spinal cord section ( a , d , g , j , m , p ; 500 μm) and amplified dorsal horn laminae I–V ( b , c , e , f , h , i , k , l , n , o , q , r ; 50 μm). Arrows indicate double immunostaining of IL-10 and each cellular biomarker. Double immunolabeled surface areas of IL-10/Iba-1 ( s ), IL-10/GFAP ( t ), and IL-10/NeuN ( u ) from the spinal dorsal horn laminae I-V indicated in white lines were quantified using the ImageJ program. Data are presented as means ± SEM ( N = 5~7 per group). * P < 0.05 vs. saline group; analyzed by unpaired and two-tailed Student’s t test

Article Snippet: Iba-1 , Merck Millipore, Darmstadt, Germany , 1:300 , 1:100.

Techniques: Expressing, Ligation, Immunofluorescence, Staining, Amplification, Double Immunostaining, Biomarker Assay, Immunolabeling, Two Tailed Test

Journal: Journal of Neuroinflammation

Article Title: Activation of GPR40 produces mechanical antiallodynia via the spinal glial interleukin-10/β-endorphin pathway

doi: 10.1186/s12974-019-1457-9

Figure Lengend Snippet: Effect of GW9508 given intrathecally on spinal β-endorphin expression on spinal microglia ( a – f ), astrocytes ( g – l ), and neurons ( m – r ) in neuropathic rats induced by spinal nerve ligation. Frozen sections of the spinal lumbar enlargements were obtained 1 h after intrathecal saline (10 μl) or GW9508 (30 μg) treatment. Immunofluorescence was double stained with β-endorphin/Iba-1, β-endorphin/GFAP, and β-endorphin/NeuN, and photomicrographs were taken from the entire spinal cord section ( a , d , g , j , m , p : 500 μm) and amplified dorsal horn laminae I–V ( b , c , e , f , h , i , k , l , n , o , q , r : 50 μm). Arrows indicate double immunostaining of β-endorphin with each cellular biomarker. Double immunolabeled surface areas of β-endorphin/Iba-1 ( s ), β-endorphin/GFAP ( t ), and β-endorphin/NeuN ( u ) from the spinal dorsal horn laminae I-V indicated in white lines were quantified using the ImageJ program. Data are presented as means ± SEM ( N = 5~7 per group). * P < 0.05 vs. saline group; analyzed by unpaired and two-tailed Student’s t test

Article Snippet: Iba-1 , Merck Millipore, Darmstadt, Germany , 1:300 , 1:100.

Techniques: Expressing, Ligation, Immunofluorescence, Staining, Amplification, Double Immunostaining, Biomarker Assay, Immunolabeling, Two Tailed Test

Journal: Journal of Neuroinflammation

Article Title: Activation of GPR40 produces mechanical antiallodynia via the spinal glial interleukin-10/β-endorphin pathway

doi: 10.1186/s12974-019-1457-9

Figure Lengend Snippet: Expression of GPR40 ( a – c ) and stimulatory effects of GW9508 on mRNA expression of IL-10, the β-endorphin precursor proopiomelanocortin (POMC) and dynorphin A precursor prodynorphin (PDYN) ( d – g ), and secretion of IL-10 and β-endorphin ( h , i ) in cultured primary microglia, astrocytes, and neurons originated from the spinal cords of 1-day-old neonatal rats. For the immunostaining study, immunofluorescence was double stained with GPR40/Iba-1, GPR40/GFAP, and GPR40/NeuN. For the stimulatory study, cultured cells and cultural medium were collected 2 h after GW9508 incubation, and the mRNA expression of IL-10, POMC, and PDYN and the peptide concentrations of IL-10 and β-endorphin were measured using quantitative real-time PCR and commercial ELISA kits, respectively. Data are presented as means ± SEM ( N = 4~7 per group). * P < 0.05 vs. control group; analyzed by unpaired and two-tailed Student’s t test

Article Snippet: Iba-1 , Merck Millipore, Darmstadt, Germany , 1:300 , 1:100.

Techniques: Expressing, Cell Culture, Immunostaining, Immunofluorescence, Staining, Incubation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Two Tailed Test