primary cultures rat cortical neurons  (BrainBits LLC)

Journal: Nutrients

Article Title: The Importance of Antioxidant Activity for the Health-Promoting Effect of Lycopene

doi: 10.3390/nu15173821

Figure Lengend Snippet: Effects of lycopene in nervous system disorders.

Article Snippet: Alzheimer′s disease , Primary cultured Sprague-Dawley rat cortical neurons , 24 h , Lycopene dissolved in tetrahydrofuran containing 0.025% butylated hydroxytoluene, added to culture medium, concentration of lycopene: 0.1, 0.5, 1, 2 or 5 μM , 1. Decreased intracellular reactive oxygen species generation and mitochondria-derived superoxide production, , , , 2016 , [ ] .

Techniques: Activity Assay, Expressing, Translocation Assay, Clinical Proteomics, Transgenic Assay, Transfection, Concentration Assay, Membrane, Activation Assay, Cell Culture, Permeability, Injection, Phospho-proteomics, Protein-Protein interactions

Journal: Neuroscience letters

Article Title: Changes in neuronal immunofluorescence in the C- versus N-terminal domains of hnRNP H following D1 dopamine receptor activation

doi: 10.1016/j.neulet.2018.07.015

Figure Lengend Snippet: DIV8 primary rat cortical neurons were treated with vehicle (Control, n = 4), 1 μM of SKF38393 (D1 dopamine receptor agonist, n = 4) and/or 10 nM of SCH23390 (D1 dopamine receptor antagonist n = 5) for 1.5 h. (A) Schematic shows hnRNP H and its quasi-RNA recognition motifs (qRRMs) and glycine-rich domains (GYR and GY). The red “Y” markings indicate the relative antibody epitopes in the N-domain or C-domain regions of hnRNP H. (B,C) ICC and intensity analysis revealed a significant increase in C-domain staining of nuclear hnRNP H following 1 μM SKF38393 [unpaired Student’s t-test, t6 = −4.94, *p = 0.003] that was blocked by co-administration of 1 μM SKF38393 and 10 nM SCH23390 [unpaired Student’s t-test, t7 = −0.34, p = 0.74]. (D) No significant difference was found in N-domain immunoreactivity of nuclear hnRNP H following 1 μM SKF38393 treatment [unpaired Student’s t-test, t6 = −1.12, p = 0.30]. (E) There was an effect of treatment on C-domain staining of hnRNP H [One-way ANOVA, F2,6 = 20.38, p < 0.001, n = 3 per condition]. SKF38393-induced increase in hnRNP H nuclear immunofluorescence was reduced in the present of the blocking peptide [post-hoc Bonferroni correction for Control vs SKF+blocking peptide: t4 = 5.849; *p = 0.004]. A significant increase in C-domain staining of nuclear hnRNP H following 1 μM SKF38393 [post-hoc Bonferroni correction for Control vs SKF: t4 = −3.454; *p = 0.025]. Black bar = Control; white bar = SKF treatment. Images were obtained on a Zeiss AxioObserver microscope with the 20X objective under uniform settings. Scale bars represent 40 μm.

Article Snippet: Primary rat cortical neuron culture and neuronal treatment Primary rat cortical neurons were dissected from neocortex of E18 Sprague-Dawley embryos (Charles River Laboratories, Wilmington, MA, USA) and grown in media as described [ 19 ].

Techniques: Control, Staining, Immunofluorescence, Blocking Assay, Microscopy

Journal: Neuroscience letters

Article Title: Changes in neuronal immunofluorescence in the C- versus N-terminal domains of hnRNP H following D1 dopamine receptor activation

doi: 10.1016/j.neulet.2018.07.015

Figure Lengend Snippet: DIV8 primary rat cortical neurons were treated with media (Control, n = 4) or 1 μM of SKF38393 (D1 dopamine receptor agonist, n = 4) for 1.5 h. (A) Schematic shows hnRNP H domains and the red “Y” marking indicates the relative antibody epitopes in the C-domain regions of hnRNP H. (B) Merged images showing co-localization of hnRNP H with NeuN (neuronal marker) and DAPI (nuclear marker). Images from individual channel are shown in Fig S5. hnRNP H staining was localized to the nucleus and no cytoplasmic localization of hnRNP H was detected. ICC and intensity analysis revealed a significant increase in C-domain staining of nuclear hnRNP H following 1 μM SKF38393 for NeuN-positive and hnRNP H-positive neurons (unpaired Student’s t-test, t6 = −5.58, *p < 0.01). (C) There was co-localized staining of hnRNP H with NeuN-positive neuronal staining and no detectable staining of hnRNP H staining in non-neuronal cells. No significant differences in percentage of NeuN/hnRNP H-positive cells [unpaired Student’s t-test, t6 = −0.052, p = 0.96] or non-NeuN cells [unpaired Student’s t-test, t6 = −0.052, p = 0.96] were noted between treatment conditions. Black bar = Control; white bar = SKF treatment. Images were obtained on a Zeiss AxioObserver microscope with the 63X objective under uniform settings. Scale bars represent 15 μm.

Article Snippet: Primary rat cortical neuron culture and neuronal treatment Primary rat cortical neurons were dissected from neocortex of E18 Sprague-Dawley embryos (Charles River Laboratories, Wilmington, MA, USA) and grown in media as described [ 19 ].

Techniques: Control, Marker, Staining, Microscopy

Journal: Neuroscience letters

Article Title: Changes in neuronal immunofluorescence in the C- versus N-terminal domains of hnRNP H following D1 dopamine receptor activation

doi: 10.1016/j.neulet.2018.07.015

Figure Lengend Snippet: DIV8 primary rat cortical neurons were treated with vehicle (Control, n = 4) or 1 μM of SKF38393 (D1 dopamine receptor agonist, n = 4) for 1.5 h. (A) qPCR analysis revealed a small decrease in Hnrnph1 mRNA (top left) [Exon 4–5: unpaired Student’s t-test, t6 = 2.866, *p = 0.03; Exon13–15: unpaired Student’s t-test, t6 = 3.367, *p=0.02] or Hnrnph2 mRNA (top right) [unpaired Student’s t-test, t6 = 2.432, p = 0.051]. (B) Immunoblot analysis of fractionated samples indicated no significant change in hnRNP H protein levels in the nucleus (Nu) or cytoplasm (Cy), as detected by either C-domain (left) or N-domain (right) antibodies. Two-way ANOVA with Cellular Compartment and Treatment as factors revealed a significant effect of Cellular Compartment [C-domain: F(1,12) = 51.45, p = 1.13 × 10−5; N-domain: F(1,12) = 123.52, p = 1.13 × 10−7], no significant effect of Treatment [C-domain: F(1,12) = 1.06, p = 0.32; N-domain: F(1,12) = 4.11, p = 0.07], and no interaction [C-domain: F(1,12) = 0.26, p = 0.62; N-domain: F(1,12) = 2.66, p = 0.13]. Successful nuclear and cytoplasmic fractionation was confirmed by CREB and HSP90. For either C- and N-domain immunoblot study: Control: n = 4, 1 μM SKF38393: n = 4. Black bar = Control; white bar = SKF treatment.

Article Snippet: Primary rat cortical neuron culture and neuronal treatment Primary rat cortical neurons were dissected from neocortex of E18 Sprague-Dawley embryos (Charles River Laboratories, Wilmington, MA, USA) and grown in media as described [ 19 ].

Techniques: Control, Western Blot, Fractionation

Journal: Cell reports

Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

doi: 10.1016/j.celrep.2019.03.093

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Long Evans rat embryonic day 19–20 mixed cortical primary neuron culture , Charles River Co. , Strain Code: 006.

Techniques: Recombinant, Cloning, Mutagenesis, Labeling, Plasmid Preparation, Software, Fractionation