Journal: MRS bulletin

Article Title: Functional imaging of brain organoids using high-density microelectrode arrays.

doi: 10.1557/s43577-022-00282-w

Figure Lengend Snippet: Figure 2. Human cerebral organoids comprise diverse cell types relevant to the study of neuronal circuit development and physiology. (a) Immunohistochemistry (IHC) revealed the presence of mature neurons (Tau/NeuN), astrocytes (GFAP), and oligodendrocytes (OLIG2) after 100 days of differentiation; nuclei were stained with DAPI (in blue). Scale bars are 50 μm. (b) Quantification of NeuN-positive cells. The ratio of the number of NeuN-positive cells to the total number of cells was quantified for both hCO lines (3 hCOs per line, for each hCO > 5000 cells). The ratio of NeuN-positive cells to total cells did not differ significantly between the two lines. (c) Single-cell RNA sequencing (scRNA- seq) was performed to quantify the cellular composition of hCOs and their comparability. The results are summarized in two pie charts with the identified cell type fractions displayed for each line. For both lines the results confirmed the presence of dopaminergic, GABAergic, glutamatergic and cholinergic neurons, neural crest cells (NCCs), radial glia (RG), astrocytes and oligodendrocytes.

Article Snippet: The following primary and secondary antibodies were used in this study: Pax6 (BioLegend, San Diego, CA, USA, #901301, 1:300), Sox2 (Sigma-Aldrich, AB5603, 1:300), FoxG1 (Abcam, ab18259, 1:200), Tuj1 (BioLegend, #801202, 1:800), Ctip2 (Abcam, ab18465, 1:200), Tbr1 (Abcam, ab31940, 1:300), MAP2 (Thermo Scientific, PA1-10,005, 1:800), Tau (Thermo Scientific, MN1000, 1:500), NeuN (Boster Bio, Pleasanton, CA, USA, M11954-3, 1:300), GFAP (Novus Biologicals, Englewood, CO, USA, NB300-141 , 1:500), goat anti-mouse IgG (H + L) highly cross-adsorbed secondary antibody, Alexa Fluor Plus 488 (Thermo Scientific, A32723, 1:400), goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary Antibody, Alexa Fluor 568 (Thermo Scientific, A11036, 1:400), goat anti-rat IgG (H + L) cross-adsorbed secondary antibody, Alexa Fluor 647 (Thermo Scientific, A21247, 1:400) and goat anti-chicken IgY (H + L) cross-adsorbed secondary antibody, Alexa Fluor Plus 647 (Thermo Scientific, A32933, 1:400).

Techniques: Immunohistochemistry, Staining, RNA Sequencing

Journal: Science Advances

Article Title: Lipid nanoparticles for mRNA delivery in brain via systemic administration

doi: 10.1126/sciadv.adw0730

Figure Lengend Snippet: Flow cytometry analysis of GFP expression ( A ) or tdTomato expression ( B ) across different brain cells [neuronal nuclei (NeuN) + for neuron, glial fibrillary acidic protein (GFAP) + for astrocyte, CD11b + for microglia, and CD31 + for BCEC]. Data are presented as means ± SD in (A) and (C) ( n = 3 biological replicates). Statistical significance was analyzed by one-way ANOVA with Tukey’s multiple comparisons test. ( C ) Schematic illustrating the delivery of Cre mRNA to activate tdTomato expression, along with the administration regimen. ( D ) Representative immunofluorescence sections of brain tissues of Ai14 mice after three administrations with PBS, MC3, or OS4T LNPs (Cre mRNA, 1 mg/kg). Scale bar, 2.5 mm. The tdTomato expression in neurons ( E ), microglia ( F ), and astrocytes ( G ) after three administrations with PBS, MC3, or OS4T LNPs (Cre mRNA, 1 mg/kg). Scale bar, 50 μm. ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The following antibodies were used: NeuN antibody (1:50 dilution, Miltenyi Biotec, clone: REA1131) for neurons; GFAP antibody (1:300 dilution, Cell Signaling Technology, cat. no. 3655S) for astrocytes; anti-Iba1 antibody (1:5000 dilution, Abcam, ab283346) and goat anti-rat IgG H&L [fluorescein isothiocyanate (FITC)] (1:1000 dilution, Abcam, ab6840) for microglia; and tdTomato (E3G5L) rabbit monoclonal antibody (1:300 dilution, Cell Signaling Technology, cat. no. 20163) and goat anti-rabbit IgG H&L [Cyanine3 (Cy3)] (1:100 dilution, Abcam, ab6939) for tdTomato.

Techniques: Flow Cytometry, Expressing, Immunofluorescence

Journal: Antioxidants & Redox Signaling

Article Title: The Neuroprotective Effect of Dimethyl Fumarate in an MPTP-Mouse Model of Parkinson's Disease: Involvement of Reactive Oxygen Species/Nuclear Factor-κB/Nuclear Transcription Factor Related to NF-E2

doi: 10.1089/ars.2016.6800

Figure Lengend Snippet: DMF treatment protects against oxidative stress. Midbrain sections were double stained for Nrf-2 (red) and NeuN (3). NeuN immunoreactivity was reduced in MPTP-injured mice (F, Q) with respect to the control group (B, Q). DMF treatment increased NeuN-positive staining at both 10 mg/kg (K, Q) and 30 mg/kg (N, K). In contrast, Nrf-2 immunoreactivity was low in the sham group (A, Q), but it increased in MPTP-injured mice (E, Q). DMF at both doses increased Nrf-2 staining (J, M, Q). Yellow spots (panels L and P) and panels D and H, are indicative of Nrf-2/NeuN co-localization. The panels (C, G, H, I and O) represent the control with DAPI to see the alive cells. Data are means ± SD of 10 mice for each group. °p < 0.05 and °°p < 0.01 versus MPTP.

Article Snippet: Sections were incubated with rabbit anti-Nrf-2 (1:100; Santa Cruz Biotechnology) or mouse monoclonal anti-neuronal nuclei (anti-NeuN) (1:100; Merck-Millipore) in a humidified chamber overnight at 37°C.

Techniques: Staining