immortalized murine microglial bv-2 cell line  (Interlab Inc)

Journal: Cells

Article Title: Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via α7 nAChR Modulation.

doi: 10.3390/cells13040309

Figure Lengend Snippet: Figure 1. α-GPC mitigates the Aβ1–42-induced detrimental effect on BV2 microglial cells. The cell viability (%) of BV2 microglial cells pre-treated for 1 h with α-GPC (1 µM) and treated for 24, 48, or 72 h with Aβ1–42 (5 µM). The vertical bars represent the means ± S.E.M. One-way ANOVA and the Bonferroni post-hoc test were used for statistical analysis. * p < 0.05.

Article Snippet: The BV2 murine microglial cell line was obtained from AcceGen (AcceGen Biotech, Fairfield, NJ, USA; Cat. ABC-TC212S; Accession Number: CVCL_0182).

Techniques:

Journal: Cells

Article Title: Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via α7 nAChR Modulation.

doi: 10.3390/cells13040309

Figure Lengend Snippet: Figure 2. Pro-inflammatory microglia were blunted by α-GPC treatment. (A) Representative images (original magnification 20×; inset 40×) of the fluorescent immunocytochemical detection of CD86 and TNF-α expression in BV2 cells pre-treated for 1 h with α-GPC (1 µM) and treated for 48 h with Aβ1–42 (5 µM) and the respective mean fluorescence intensity (MFI) analysis (A’), the boxes in the fourth column represent the selected magnified area shown in the fifth column (inset). (B) Western blot for TNF-α protein expression in BV2 cells and the respective densitometric analysis (B’). The data are expressed as means ± S.E.M. One-way ANOVA and the Bonferroni post-hoc test were used to determine statistical significance. * p < 0.05.

Article Snippet: The BV2 murine microglial cell line was obtained from AcceGen (AcceGen Biotech, Fairfield, NJ, USA; Cat. ABC-TC212S; Accession Number: CVCL_0182).

Techniques: Expressing, Fluorescence, Western Blot

Journal: Cells

Article Title: Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via α7 nAChR Modulation.

doi: 10.3390/cells13040309

Figure Lengend Snippet: Figure 3. α-GPC contributes to the switching of microglia toward a pro-inflammatory phenotype. (A) Representative images (original magnification 20×; inset 40×) of the fluorescent immunocy- tochemical detection of CD68 and IL-10 expression in BV2 cells pre-treated for 1 h with α-GPC (1 µM) and treated for 48 h with Aβ1–42 (5 µM) and the respective mean fluorescence intensity (MFI) analysis (A’), the boxes in the fourth column represent the selected magnified area shown in the fifth column (inset). (B) Western blot for IL-10 protein expression in BV2 cells and the respective densitometric analysis (B’). The data are expressed as means ± S.E.M. One-way ANOVA and the Bonferroni post-hoc test were used to determine statistical significance. * p < 0.05.

Article Snippet: The BV2 murine microglial cell line was obtained from AcceGen (AcceGen Biotech, Fairfield, NJ, USA; Cat. ABC-TC212S; Accession Number: CVCL_0182).

Techniques: Expressing, Fluorescence, Western Blot

Journal: Cells

Article Title: Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via α7 nAChR Modulation.

doi: 10.3390/cells13040309

Figure Lengend Snippet: Figure 4. α7 nAchR is modulated by α-GPC treatment. (A) The fluorescent immunocytochemistry detection (original magnification 20×) of α7 nAChR in BV2 cells pre-treated with α-GPC (1 µM) and treated for 48 h with Aβ1–42 (5 µM) and the respective mean fluorescence intensity (MFI) analysis (A’). (B) Western blot for α7 nAChR protein expression in BV2 cells and the respective densitometric analysis (B’). The data are expressed as means ± S.E.M. One-way ANOVA and the Bonferroni post-hoc test were used to determine statistical significance. * p < 0.05.

Article Snippet: The BV2 murine microglial cell line was obtained from AcceGen (AcceGen Biotech, Fairfield, NJ, USA; Cat. ABC-TC212S; Accession Number: CVCL_0182).

Techniques: Immunocytochemistry, Fluorescence, Western Blot, Expressing

Journal: Cells

Article Title: Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via α7 nAChR Modulation.

doi: 10.3390/cells13040309

Figure Lengend Snippet: Figure 5. Effect of α-bungarotoxin on acetylcholine-induced [Ca2+]i increase and acetylcholine- induced inward current in microglial BV2 cells. (A) Representative traces and quantification (B) for the effect of acetylcholine (Ach, 1 µM) alone or in the presence of α-bungarotoxin (10 nM) on [Ca2+]i, expressed as the ∆% increase peak over basal values (N = 40 cells for Ach and N = 35 cells for Ach + α-BTX). On the left, the representative brightfield and pseudocolor images of Fura-2 loaded BV2 cells are shown. (C) Representative current traces in response to Ach (1 µM) alone or in the presence of α-BTX (10 nM), and the quantification of current amplitude (D). The values are expressed as the mean ± SEM of three independent experimental sessions (N = 8 cells for Ach and N = 10 cells for Ach + α-BTX). * p < 0.05.

Article Snippet: The BV2 murine microglial cell line was obtained from AcceGen (AcceGen Biotech, Fairfield, NJ, USA; Cat. ABC-TC212S; Accession Number: CVCL_0182).

Techniques:

Journal: Cells

Article Title: Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via α7 nAChR Modulation.

doi: 10.3390/cells13040309

Figure Lengend Snippet: Figure 6. Effect of α-GPC on [Ca2+]i increase and the α7-nAChR-encoded inward current in microglial BV2 cells. (A) Representative traces and quantification (B) for the effect of different concentrations of α-GPC (0.01–1 µM) alone or α-GPC (1 µM) + α-BTX (10 nM) on [Ca2+]i, expressed as the ∆% increase peak over basal values (N = 35 cells for α-GPC and N = 30 cells for α-GPC + α-BTX). On the left of each panel, the representative brightfield and pseudocolor images of Fura-2 loaded BV2 cells are shown. * p < 0.05 vs. control (basal values) and 0.0001 µM; ** p < 0.05 vs. control and 0.01 µM; *** p < 0.05 vs. all. (C) Representative traces of Ach-like currents elicited by α-GPC (1 µM) alone or + α-BTX (10 nM), and the quantification of current amplitude (D). The values are expressed as the mean ± SEM of three independent experimental sessions (N = 9 cells for α-GPC and N = 12 cells for α-GPC + α-BTX). (E) Quantification of the slow deactivation induced by α-GPC. * p < 0.05.

Article Snippet: The BV2 murine microglial cell line was obtained from AcceGen (AcceGen Biotech, Fairfield, NJ, USA; Cat. ABC-TC212S; Accession Number: CVCL_0182).

Techniques: Control

Journal: Cells

Article Title: Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via α7 nAChR Modulation.

doi: 10.3390/cells13040309

Figure Lengend Snippet: Figure 7. α-BTX restores the expression of pro-inflammatory molecules. (A) Representative images (original magnification 20×; inset 40×) of the fluorescent immunocytochemical detection of CD86 and TNF-α expression in BV2 cells pre-treated for 1 h with α-GPC (1 µM) and treated for 48 h with Aβ1–42 (5 µM)/α-BTX (100 nM) and the respective mean fluorescence intensity (MFI) analysis (A’), the boxes in the fourth column represent the selected magnified area shown in the fifth column (inset). The data are expressed as means ± S.E.M. Differences between the groups were considered significant at * p < 0.05 (One-way ANOVA followed by the Bonferroni post-hoc test).

Article Snippet: The BV2 murine microglial cell line was obtained from AcceGen (AcceGen Biotech, Fairfield, NJ, USA; Cat. ABC-TC212S; Accession Number: CVCL_0182).

Techniques: Expressing, Fluorescence

Journal: Cells

Article Title: Taming Microglia in Alzheimer's Disease: Exploring Potential Implications of Choline Alphoscerate via α7 nAChR Modulation.

doi: 10.3390/cells13040309

Figure Lengend Snippet: Figure 8. α-BTX inhibits the anti-inflammatory effects of α-GPC. (A) Representative images (original magnification 20×; inset 40×) of the fluorescent immunocytochemical detection of CD68 and IL-10 expression in BV2 cells pre-treated for 1 h with α-GPC (1 µM) and treated for 48 h with Aβ1–42 (5 µM)/α-BTX (100 nM) and the respective mean fluorescence intensity (MFI) analysis (A’), the boxes in the fourth column represent the selected magnified area shown in the fifth column (inset). The data are expressed as means ± S.E.M. Differences between the groups were considered significant at * p < 0.05 (One-way ANOVA followed by the Bonferroni post-hoc test).

Article Snippet: The BV2 murine microglial cell line was obtained from AcceGen (AcceGen Biotech, Fairfield, NJ, USA; Cat. ABC-TC212S; Accession Number: CVCL_0182).

Techniques: Expressing, Fluorescence