Journal: Cell reports

Article Title: GLP-1 Receptor Agonist NLY01 Reduces Retinal Inflammation and Neuron Death Secondary to Ocular Hypertension

doi: 10.1016/j.celrep.2020.108271

Figure Lengend Snippet: C57BL6/J (WT) mice were injected either with microbeads (“Bead,” left eye) into the AC to increase IOP or with BSS (right eye). (A–C) ELISA measurements of IL-1α (A), TNF-α (B), or C1q (C) protein levels in whole neurosensory retina at 1, 2, 3, 5, 7, 14, 28, and 42 days post-injection. Statistical test compared BSS-injected eyes to microbead-injected eyes at the same time points (n = 5 eyes per time point per condition). (D–F) CD11b + CD11c − and CD11b + CD11c + cells were isolated from neurosensory retina at 1, 2, 3, 5, 7, 14, 28, and 42 days post-injection. Il1a (D), Tnf (E), and C1qa (F) mRNA levels were measured by qPCR in both cell populations. qPCR measurements in microbead-injected eyes were normalized to the contralateral BSS-injected eyes. Statistical tests compared microbead-injected CD11b + CD11c + cells and microbead-injected CD11b + CD11c − cells at the same time points (n = 5 eyes per time point per condition). All data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, Mann-Whitney U test. See also – .

Article Snippet: The following isolation steps were performed: (1) positive selection for ACSA2 (astrocyte and Muller cell enriched, 130-097-678, Miltenyi Biotec), (2) the remaining negative selection pool was then subjected to positive selection for CD11b (microglia and macrophage enriched, 130-093-636 Miltenyi Biotec), and (3) the CD11b + cells were then selected against CD11c (130-125-835, Miltenyi Biotec).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Isolation, MANN-WHITNEY

Journal: Cell reports

Article Title: GLP-1 Receptor Agonist NLY01 Reduces Retinal Inflammation and Neuron Death Secondary to Ocular Hypertension

doi: 10.1016/j.celrep.2020.108271

Figure Lengend Snippet: C57BL6/J (WT) mice were injected either with microbeads (“Bead,” left eye) to increase IOP or with BSS (right eye). Following intraocular injections, mice were randomized to twice-weekly subcutaneous NLY01 (5 mg/kg per injection) or normal saline. Mice were euthanized 14 days post-injection. (A–F) CD11b + CD11c − and CD11b + CD11c + cells were isolated from neurosensory retina. qPCR was performed to measure Il1a (A and D), Tnf (B and E), and C1qa (C and F) mRNA levels in each population (n = 5 eyes per condition). (G) qPCR measurements of pan-reactive, A1-specific, and A2 specific transcripts from ACSA2 + cells 14 days post-injection (n = 5 eyes per condition). (H) qPCR measurement of C3 mRNA levels in ACSA2 + cells. (n = 5 eyes per condition) All data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to BSS NSS (Mann-Whitney U test). See also , , and .

Article Snippet: The following isolation steps were performed: (1) positive selection for ACSA2 (astrocyte and Muller cell enriched, 130-097-678, Miltenyi Biotec), (2) the remaining negative selection pool was then subjected to positive selection for CD11b (microglia and macrophage enriched, 130-093-636 Miltenyi Biotec), and (3) the CD11b + cells were then selected against CD11c (130-125-835, Miltenyi Biotec).

Techniques: Injection, Saline, Isolation, MANN-WHITNEY

Journal: Cell reports

Article Title: GLP-1 Receptor Agonist NLY01 Reduces Retinal Inflammation and Neuron Death Secondary to Ocular Hypertension

doi: 10.1016/j.celrep.2020.108271

Figure Lengend Snippet: C57BL6/J (WT) mice were injected either with microbeads (“Bead,” left eye) to increase IOP or with BSS (right eye). Following injections, mice were randomized to twice-weekly subcutaneous NLY01 (5 mg/kg per injection) or normal saline. Mice were euthanized 42 days post-injection. (A–F) CD11b + CD11c − and CD11b + CD11c + cells were isolated from neurosensory retina. qPCR was performed to measure Il1a (A and D), Tnf (B and E), and C1qa (C and F) mRNA levels in each population (n = 5 eyes per condition). (G) qPCR measurements of pan-reactive, A1-specific, and A2 specific transcripts from ACSA2 + cells 42 days post-injection (n = 5 eyes per condition). (H) qPCR measurement of C3 mRNA levels in ACSA2 + cells (n = 5 eyes per condition). All data presented as mean ± SEM. Mann-Whitney U test, **p < 0.01, ***p < 0.001, ****p < 0.0001 versus BSS NSS. See also , , and .

Article Snippet: The following isolation steps were performed: (1) positive selection for ACSA2 (astrocyte and Muller cell enriched, 130-097-678, Miltenyi Biotec), (2) the remaining negative selection pool was then subjected to positive selection for CD11b (microglia and macrophage enriched, 130-093-636 Miltenyi Biotec), and (3) the CD11b + cells were then selected against CD11c (130-125-835, Miltenyi Biotec).

Techniques: Injection, Saline, Isolation, MANN-WHITNEY

Journal: Cell reports

Article Title: GLP-1 Receptor Agonist NLY01 Reduces Retinal Inflammation and Neuron Death Secondary to Ocular Hypertension

doi: 10.1016/j.celrep.2020.108271

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The following isolation steps were performed: (1) positive selection for ACSA2 (astrocyte and Muller cell enriched, 130-097-678, Miltenyi Biotec), (2) the remaining negative selection pool was then subjected to positive selection for CD11b (microglia and macrophage enriched, 130-093-636 Miltenyi Biotec), and (3) the CD11b + cells were then selected against CD11c (130-125-835, Miltenyi Biotec).

Techniques: Recombinant, Saline, Injection, Sterility, Electron Microscopy, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Software, Real-time Polymerase Chain Reaction

Journal: Journal of neurochemistry

Article Title: Differential susceptibility of astrocytic and neuronal function to 3-chloropropanediol in the rat inferior colliculus.

doi: 10.1111/j.1471-4159.2010.07138.x

Figure Lengend Snippet: Fig. 2 Time course of the loss of immuno-positive astrocytes and microglia from the inferior colliculi over 0–72 h survival after 3-CPD. The number of GFAP-positive astrocytes (-j-) and CD11b-positve microglia (-h-) were normalized to a percentage of that found in controls. Points represent mean ± SE. Statistically significant results are indicated as *p < 0.05 or **p < 0.01.

Article Snippet: Glial fibrillary acidic protein (GFAP) positive astrocytes were detected using a mouse monoclonal antibody (G3893, Sigma) at a dilution of 1 in 1000; and CD11b positive microglia detected using a mouse monoclonal antibody (MC275R, Serotec, Kidlington, Oxfordshire, UK) at a dilution of 1 in 2000.

Techniques: