Journal: Journal of Pain Research

Article Title: The Interaction Between Spinal PDGFRβ and μ Opioid Receptor in the Activation of Microglia in Morphine-Tolerant Rats

doi: 10.2147/JPR.S255221

Figure Lengend Snippet: Expression of Iba1 in spinal cord of rats. ( A ) Thermal pain threshold of rats was assessed using the percentage of maximal possible antinociceptive effect (%MPE) according to the tail-flick latency. The %MPE in rats receiving morphine (10 μg, twice daily, intrathecally) on days 5 and 7 were decreased compared with the baseline on day 1. *** P <0.001, **** P <0.0001, vs NS rats. ( B, C ) The expressions of Iba1 mRNA ( B ) and protein ( C ) were significantly increased in morphine-tolerant rats measured by real-time PCR and Western blots, respectively. ** P <0.01, vs NS rats. Values represent mean±SEM. n=6 in each group. Abbreviations: NS, normal saline; MT, morphine tolerance; %MPE, the percentage of maximal possible antinociceptive effect.

Article Snippet: The membranes were blocked with 5% bovine serum albumin for 2 hours at room temperature and incubated overnight at 4°C with mouse anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody (1:200; Santa Cruz, USA), rabbit anti-p-JNK (Thr180/Tyr183) (1:1000; CST, USA), rabbit anti-phospho-PDGFRβ (1:1000; CST), rabbit anti-p-c-JUN (1:1000; CST), or rabbit anti-GAPDH (1:2000; Aspen, China).

Techniques: Expressing, Tail Flick Test, Real-time Polymerase Chain Reaction, Western Blot, Saline

Journal: Journal of Pain Research

Article Title: The Interaction Between Spinal PDGFRβ and μ Opioid Receptor in the Activation of Microglia in Morphine-Tolerant Rats

doi: 10.2147/JPR.S255221

Figure Lengend Snippet: The effect of PDGFRβ agonist PDGF-BB on Iba1 expression. ( A ) PDGF-BB had no influence on %MPE of rats. ( B – D ) The phosphorylation of PDGFRβ ( B ) and the expressions of Iba1 mRNA ( C ) and protein ( D ) were significantly increased in rats received PDGF-BB measured by real-time PCR and Western blots, respectively. * P <0.05, ** P <0.01 vs PBS rats. ( E ) Rats were intrathecally injected with PDGF-BB or PBS once daily for 4 days, followed by a single dose of morphine on day 5. The %MPE in rats receiving PDGF-BB and morphine was decreased. ** P <0.01, vs PBS+Morphine rats. ( F ) The %MPE in rats receiving imatinib 30 minutes before morphine administration were higher than those in morphine-tolerant rats from day 5 to 7. ** P <0.01, vs Morphine+NS rats. ( G ) Pretreatment with imatinib inhibited the increased expression of Iba1 induced by morphine measured by Western blots. * P <0.05, vs NS+NS rats; # P <0.05, vs Morphine+NS rats. Values represent mean±SEM. n=6 in each group. Abbreviations: NS, normal saline; PBS, phosphate buffered saline; %MPE, the percentage of maximal possible antinociceptive effect.

Article Snippet: The membranes were blocked with 5% bovine serum albumin for 2 hours at room temperature and incubated overnight at 4°C with mouse anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody (1:200; Santa Cruz, USA), rabbit anti-p-JNK (Thr180/Tyr183) (1:1000; CST, USA), rabbit anti-phospho-PDGFRβ (1:1000; CST), rabbit anti-p-c-JUN (1:1000; CST), or rabbit anti-GAPDH (1:2000; Aspen, China).

Techniques: Expressing, Phospho-proteomics, Real-time Polymerase Chain Reaction, Western Blot, Injection, Saline

Journal: Journal of Pain Research

Article Title: The Interaction Between Spinal PDGFRβ and μ Opioid Receptor in the Activation of Microglia in Morphine-Tolerant Rats

doi: 10.2147/JPR.S255221

Figure Lengend Snippet: The effect of MOR antagonist naloxone on activations of PDGFRβ and microglia. ( A ) The %MPE in rats receiving naloxone 30 minutes before morphine administration were higher than those in morphine-tolerant rats from day 5 to 7. * P <0.05, *** P <0.001, vs Morphine+DMSO rats. ( B, C ) Pretreatment with naloxone inhibited the phosphorylation of PDGFRβ ( B ) and the increased expression of Iba1 ( C ) induced by morphine measured by Western blots. * P <0.05, ** P <0.01, vs NS+DMSO rats; # P <0.05, vs Morphine+DMSO rats. Values represent mean±SEM. n=6 in each group. Abbreviations: NS, normal saline; DMSO, dimethyl sulfoxide; %MPE, the percentage of maximal possible antinociceptive effect.

Article Snippet: The membranes were blocked with 5% bovine serum albumin for 2 hours at room temperature and incubated overnight at 4°C with mouse anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody (1:200; Santa Cruz, USA), rabbit anti-p-JNK (Thr180/Tyr183) (1:1000; CST, USA), rabbit anti-phospho-PDGFRβ (1:1000; CST), rabbit anti-p-c-JUN (1:1000; CST), or rabbit anti-GAPDH (1:2000; Aspen, China).

Techniques: Phospho-proteomics, Expressing, Western Blot, Saline

Journal: Journal of Pain Research

Article Title: The Interaction Between Spinal PDGFRβ and μ Opioid Receptor in the Activation of Microglia in Morphine-Tolerant Rats

doi: 10.2147/JPR.S255221

Figure Lengend Snippet: Involvement of JNK signaling in MOR-induced PDGFRβ activation in morphine tolerance. ( A ) Pretreatment with naloxone 30 minutes before morphine administration inhibited the phosphorylation of JNK induced by morphine measured by Western blots. ** P <0.01, vs NS+DMSO rats; # P <0.05, vs Morphine+DMSO rats. ( B ) The %MPE in rats receiving JNK inhibitor SP600125 30 minutes before morphine administration were higher than those in morphine-tolerant rats from day 5 to 7. ** P <0.01, vs Morphine+DMSO rats. ( C–E ) Pretreatment with SP600125 reduced increased expressions of p-c-Jun ( C ), p-PDGFRβ ( D ), and Iba1 ( E ) induced by morphine measured by Western blots. * P <0.05, *** P <0.001, vs NS+DMSO rats; # P <0.05, ## P <0.01, vs Morphine+DMSO rats. ( F ) Pretreatment with imatinib had no influence on the increased expression of p-JNK induced by morphine measured by Western blots. ** P <0.01, vs NS+NS rats. Values represent mean±SEM. n=6 in each group. Abbreviations: NS, normal saline; DMSO, dimethyl sulfoxide; %MPE, the percentage of maximal possible antinociceptive effect.

Article Snippet: The membranes were blocked with 5% bovine serum albumin for 2 hours at room temperature and incubated overnight at 4°C with mouse anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody (1:200; Santa Cruz, USA), rabbit anti-p-JNK (Thr180/Tyr183) (1:1000; CST, USA), rabbit anti-phospho-PDGFRβ (1:1000; CST), rabbit anti-p-c-JUN (1:1000; CST), or rabbit anti-GAPDH (1:2000; Aspen, China).

Techniques: Activation Assay, Phospho-proteomics, Western Blot, Expressing, Saline

Journal: CNS Neuroscience & Therapeutics

Article Title: Pleiotrophin Overexpression Reduces Adolescent Ethanol Consumption and Modulates Ethanol‐Induced Glial Responses and Changes in the Perineuronal Nets in the Mouse Hippocampus

doi: 10.1111/cns.70159

Figure Lengend Snippet: Effects of IAE and Ptn overexpression on microglial responses in the adolescent hippocampus. Photomicrographs are from Iba1‐immunostained hippocampal sections of Ptn‐ Wt and Ptn ‐Tg male and female mice. Dashed lines indicate the outline of the granule cell layer of the hippocampal dentate gyrus. In the lower left corner, we present a magnified image of a single microglia cell (a). Graphs represent data (mean ± S.E.M.) from the quantification of Iba1 + number of cells (b) in the granular cell layer. Iba 1 + average cell size (c) and circularity (d) are also shown. Graphs depicting Iba1 + body size (e), ramification length (f), and ramification complexity of the Iba1+ cells (g) in this area are also presented. #### p < 0.0001 for significant effect of genotype. $$ p < 0.01; $$$$ p < 0.0001" replacing $$$ p < 0.001 for significant effect of drink. ^ p < 0.05; ^^^^ p < 0.0001 for significant effect of drink excluding sex variable. * p < 0.05; **** p < 0.0001 for significant effect of genotype excluding sex variable. ££££ p < 0.0001 vs. Female Water Ptn ‐Tg. Scale bar = 100 μm.

Article Snippet: In addition, to assess glial responses, DG sections were incubated overnight at 4°C with mouse anti‐glial fibrillary acidic protein (GFAP; Millipore, Madrid, Spain; 1:1000) and rabbit anti‐ionized calcium‐binding adaptor molecule 1 (Iba1, Wako, Osaka, Japan; 1:1000) antibodies, followed by a 30‐min incubation with the corresponding Alexa‐Fluor‐555 and Alexa‐Fluor‐488 secondary antibodies (Invitrogen, Waltham, MA, USA; 1:500).

Techniques: Over Expression

Journal: PLOS Neglected Tropical Diseases

Article Title: Vertical transmission of Leishmania donovani with placental degeneration in the pregnant mouse model of visceral leishmaniasis

doi: 10.1371/journal.pntd.0012650

Figure Lengend Snippet: (A) The expression of CD206, MRP14, B220 and CD3 in the placentas. Ld infection decreased CD206 + MΦs and MRP14 + MΦs, and increased CD3 + T cells in the placentas. (B) Consistent localization of CD3, ICAM1, and VCAM1 in serial placental sections. (C) cNK and uNK cells in DBA/PAS-stained placental sections. Open arrowhead indicates cNK cells (DBA - PAS + ), and closed arrowhead indicates uNK cells (DBA + PAS + ). The number of cells was counted in 5 random microscopic fields of placental labyrinth zone at 400 × magnification (n = 5-21). Bars, 50 μm. Means ± SE are presented. P values for Student’s t test are shown.

Article Snippet: After blocking, rabbit anti-kinetoplastid membrane protein-11 (KMP11) antibody, rabbit anti-CD206 antibody (Cell Signal Technology, Danvers, MA, USA), goat anti-MRP14 antibody (Santa Cruz Biotechnology, Santa Cruz, CA), goat anti-CD3 antibody (Santa Cruz Biotechnology), rat anti-B220 antibody (BD Biosciences, San Jose, CA), goat anti-ICAM1 antibody (Santa Cruz Biotechnology) or goat anti-VCAM1 antibody (Santa Cruz Biotechnology) was applied to the serial sections of tissues.

Techniques: Expressing, Infection, Staining