Journal: Epilepsia

Article Title: Complex alterations in microglial M1/M2 markers during the development of epilepsy in two mouse models.

doi: 10.1111/epi.12960

Figure Lengend Snippet: Figure 1. Microglial-specific expression of M1 and M2 markers peak at the acute (3 days post-SE) but not early chronic (21 days post-SE) time point in pilocarpine SE forebrains as determined by flow cytometry. (A) M1-associated cytokines were collectively increased at the 3-day time point in SE compared to No-SE animals, including TNFa, IL-1b, IL-6, and IL-12, but returned to control levels after 21 days. (B) M2- associated cellular markers, Arg1, Ym1, IL-4, and IL-10 were all increased 3 days, but not 21 days, after SE. Only Ym1 remained elevated in SE mice at 21 days post- SE. (C) Representative median fluorescent intensity (MFI) curves of M1/M2 marker expression on microglial cells comparing naive (dotted line), No-SE (solid line, open), and SE (shaded fill) animals determined by flow cytometry at the 3-day post-SE time point. Data shown are normalized to naive controls (n = 8 per group for 3 days; n = 12 per group for 21 days after SE). ***p < 0.001, ****p < 0.0001. Mean fold change SEM shown. Two-way analysis of variance (ANOVA) with Bonferroni posttest used for statistical analysis of all data sets. Epilepsia ILAE

Article Snippet: Intracellular Arg1 (Fluorescein, IC5868F; R&D Systems, Minneapolis, MN, U.S.A.) and Ym1 (nonconjugated, MAB2446; R&D Systems; secondary – PE conjugated) were visualized after cell permeabilization.

Techniques: Expressing, Cytometry, Control, Marker

Journal: Epilepsia

Article Title: Complex alterations in microglial M1/M2 markers during the development of epilepsy in two mouse models.

doi: 10.1111/epi.12960

Figure Lengend Snippet: Figure 2. mRNA expression of M1 and M2 phenotypic markers at different stages throughout the pilocarpine model in forebrain (acute 3 days post-SE) and the hippocampal formation (early and late chronically epileptic time points) largely peak early after SE. (A) M1-associated markers TNFa, IL-1b, CD16, and CD11b and the astrocytic marker GFAP were significantly increased in forebrain mRNA of SE animals compared to No-SE controls at the acute post-SE time point. At the early chronic time point, IL-1b, CD11b, and GFAP remained elevated above No-SE control levels in the hippocampal formation, despite there being no obvious changes in whole cortical samples (data not shown). TNFa mRNA levels, however, were significantly reduced at the early chronic stage in hippocampus. No significant changes to any M1 markers were observed in hippocampal mRNA extracts in the late chronic phase (n = 5–9 per group). (B) M2-associated markers Arg1, Ym1, and IL-4 were significantly increased in SE animals compared to No-SE controls, whereas FIZZ-1 and CD206 showed signifi- cant reductions in expression in SE animals at the acute time point in forebrain (n = 5–9 per group). Together this indicates a heteroge- neous M1/M2 microglial activation state acutely after pilocarpine-induced SE. M2-associated Ym1 was the only marker for which mRNA amounts decreased in the hippocampal formation at the early chronic time point (n = 12 per group). No significant changes to any other markers were found in either the early or late chronic phases in hippocampal mRNA extracts. Data were expressed relative to house- keeping gene mRNA expression. Mean SEM shown for each data set. **p < 0.01, ***p < 0.001, ****p < 0.0001. Independent unpaired student’s t-tests used for statistical analysis of each data set. Epilepsia ILAE

Article Snippet: Intracellular Arg1 (Fluorescein, IC5868F; R&D Systems, Minneapolis, MN, U.S.A.) and Ym1 (nonconjugated, MAB2446; R&D Systems; secondary – PE conjugated) were visualized after cell permeabilization.

Techniques: Expressing, Marker, Control, Activation Assay

Journal: Epilepsia

Article Title: Complex alterations in microglial M1/M2 markers during the development of epilepsy in two mouse models.

doi: 10.1111/epi.12960

Figure Lengend Snippet: Figure 3. Phasic increased hippocampal mRNA expression of M1 and M2 markers in i.h. kainate post-SE mice over disease progression reveal early and late marker expression. (A) M1-associated TNFa, IL-1b, and CD11b were all increased in SE mice compared to No-SE controls at the acute and late chronically epileptic time points. No changes in M1 expression were observed at the early chronic time point (n = 6– 12 per group). Antigen presentation M1-associated marker CD16 was increased in SE animals only acutely post-SE. Similarly astrocytic activation marker GFAP was only significantly increased in SE mice at the acute time point, and did show a trend of increased expression in the late chronic epileptic period. (B) Selected M2 markers were increased in SE animals in the i.h. kainate model. No significant increases in any M2-associated marker were measured at either the acute or early chronic time points; however, oxidative stress regula- tor enzyme Arg1, mannose receptor CD206, and anti-inflammatory cytokine IL-10 were increased in SE mice in the late chronic time point in this model. No significant changes to Ym1 or IL-4 expression were observed at this same time point. Mean SEM were shown for each group. n = 5–12 per group. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Two-way ANOVA with Bonferroni post- test used for statistical analysis of all data sets. Epilepsia ILAE

Article Snippet: Intracellular Arg1 (Fluorescein, IC5868F; R&D Systems, Minneapolis, MN, U.S.A.) and Ym1 (nonconjugated, MAB2446; R&D Systems; secondary – PE conjugated) were visualized after cell permeabilization.

Techniques: Expressing, Biomarker Discovery, Marker, Immunopeptidomics, Activation Assay

Journal: bioRxiv

Article Title: Resveratrol reduces COMPopathy in mice through activation of autophagy

doi: 10.1101/2020.10.26.355628

Figure Lengend Snippet: Treatments started at 1 or 2 or 3 weeks after birth and stopped at 4 weeks and evaluated for mutant COMP pathology. Ten mice were included in each treatment group. T reatments starting at birth or 1 week of age produced the best therapeutic outcomes ( K, L ). Later therapy dampened but did not eliminate the disease progression and did not restore DNA proliferation ( C ). Resveratrol treatment times are shown in the shaded boxes. Control (C57BL\6) ( A, G, M, S, Y, EE ) and MT-COMP with no treatment ( B, H, N, T, Z, FF ) resveratrol treatment for 1 ( C, I, O, U, AA, GG ), 2 ( D, J, P, V, BB, HH ), 3 ( E, K, Q, W, CC, II ) or 4 ( F, L, R, X, DD, JJ ) weeks and growth plates at 4 weeks were evaluated for human COMP retention, IL-16 and YM1, TUNEL (cell death), PCNA (cell proliferation) and mTORC1 signaling (pS6). Quantification of TUNEL positive chondrocytes for different treatment periods are shown in KK and summary of findings in LL . Bars in KK = means with standard deviation. Resveratrol treatment decreased intracellular MT-COMP ( C ) and IL-16 and YM1 inflammation ( I, O ) after one week of treatment compared to untreated MT-COMP growth plates ( H, N ) and mTORC1 signaling was substantially reduced after 3 weeks of treatment ( II, JJ ). The relative change of MT-COMP pathology on treatment period is shown in LL .

Article Snippet: Immunostaining was performed by incubating different sections overnight at 4 °C using the following antibodies: human COMP (Abcam Cambridge, MA ab11056-rat 1:100), pS6 (1:200 2215S rabbit polyclonal Cell Signaling Technology), PI3K-1 (abcam-ab 225720-1:200), SIRT-1 (abcam: ab 32441 - 1:100), pAMPK (Invitrogen: PA537821-1:100), interleukin 16 (IL-16) (Santa Cruz Biotechnology; sc-7902, 1:100), YM1/ECF-L (Stem Cell Technologies; 01404, 1:100) and PCNA (abcam ab92552 - 1:200).

Techniques: Mutagenesis, Produced, TUNEL Assay, Standard Deviation

Journal: bioRxiv

Article Title: Resveratrol reduces COMPopathy in mice through activation of autophagy

doi: 10.1101/2020.10.26.355628

Figure Lengend Snippet: Treatments started at 1 or 2 or 3 weeks after birth and stopped at 4 weeks and evaluated for mutant COMP pathology. Each treatment group included 10 mice. Aspirin treatments are shown in the shaded boxes. Treatments starting at birth or 1 week of age produced the best therapeutic outcomes ( K, L ), later therapy mitigated but did not eliminate the disease progression ( C ). Control (C57BL\6) ( A, G, M, S, Y, EE ) and MT-COMP with no treatment ( B, H, N, T, Z, FF ) resveratrol treatment for 1 ( C, I, O, U, AA, GG ), 2 ( D, J, P, V, BB, HH ), 3 ( E, K, Q, W, CC, II ) or 4 ( F, L, R, X, DD, JJ ) weeks and growth plates at 4 weeks were evaluated for human COMP retention, IL-16 and YM1, TUNEL (cell death), PCNA and mTORC1 signaling. Quantification of TUNEL positive chondrocytes for different treatment periods are shown in KK . Bars = means with standard deviation. Aspirin treatment decreased intracellular MT-COMP after two weeks of treatment ( D ) and decreased in IL-16 and YM1 inflammation after one week of treatment ( I, O ) compared to ( H, N). Aspirin decreased cell death after 2 weeks of treatment ( U ), mTORC1 signaling after 3 weeks of treatment ( JJ ) and increased proliferation after 3 weeks of treatment ( CC ). The relative change of MT-COMP pathology on treatment period is shown in LL .

Article Snippet: Immunostaining was performed by incubating different sections overnight at 4 °C using the following antibodies: human COMP (Abcam Cambridge, MA ab11056-rat 1:100), pS6 (1:200 2215S rabbit polyclonal Cell Signaling Technology), PI3K-1 (abcam-ab 225720-1:200), SIRT-1 (abcam: ab 32441 - 1:100), pAMPK (Invitrogen: PA537821-1:100), interleukin 16 (IL-16) (Santa Cruz Biotechnology; sc-7902, 1:100), YM1/ECF-L (Stem Cell Technologies; 01404, 1:100) and PCNA (abcam ab92552 - 1:200).

Techniques: Mutagenesis, Produced, TUNEL Assay, Standard Deviation

Journal: British Journal of Pharmacology

Article Title: Boosting phagocytosis and anti‐inflammatory phenotype in microglia mediates neuroprotection by PPARγ agonist MDG548 in Parkinson's disease models

doi: 10.1111/bph.14214

Figure Lengend Snippet: MDG548 modified the production of TNF‐α, MRC1 and iNOS in MMGT12 cells. (A) MDG548 effect at the doses of 10 and 50 μM (MDG10 and MDG50, respectively). (B‐F) Effect of LPS alone or in association with MDG548 10 μM, added 2 h after LPS (LPS + MDG10s) or at the same time point (LPS + MDG10c). TNF‐α, IL‐1β, IL‐10, MRC1 and Ym1 were measured by elisa. (G) Effect of MDG548, LPS alone or in association with MDG548 on iNOS expression. iNOS was measured by immunohistochemistry. ^ P < 0.05, significantly different from control cells; * P < 0.05, significantly different from LPS alone‐treated cells; # P < 0.05, significantly different from LPS + MDG10s‐treated cells.

Article Snippet: Mouse http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5074 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4974 ( https://www.thermofisher.com/us/en/home.html , Waltham, MA, USA), Ym1 (EIAab Science, Wuhan, China), MRC1/CD206 (Cloud‐Clone Corp., Katy, TX, USA) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4975 (Sigma‐Aldrich Corp., St. Louis, MO, USA) were assessed by sandwich elisa according to the manufacturer's instructions.

Techniques: Modification, Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry