Journal: Iubmb Life

Article Title: Tyrosine and Phenylalanine Activate Neuronal DNA Repair but Exhibit Opposing Effects on Global Transcription and Adult Female Mice Are Resilient to TyrRS / YARS1 Depletion

doi: 10.1002/iub.70030

Figure Lengend Snippet: cis ‐RSV and trans ‐RSV have opposite effects on Aβ‐induced neurite degeneration and DNA damage response. (A) cis‐ RSV and trans ‐RSV have opposite effects on Aβ‐induced TyrRS depletion. Representative spectral images (scale bar, 20 μm) and quantification of TyrRS protein in rat cortical neurons (DIV 10) treated with cis ‐RSV or trans ‐RSV, alone or in combination with nMAβ 42 (50 nM) for 16 h. (B) cis ‐RSV prevents, while trans‐RSV exacerbates nMAβ42‐induced neurite degeneration. Representative images (scale bar, 20 μm) of cortical neurons pre‐treated with cis ‐RSV and trans ‐RSV (50 μM) for 16 h, followed by nMAβ 42 (50 nM) exposure for 24 h. Neurons were stained with MAP2 (neurite marker, magenta) and DAPI (nuclear marker, blue). Neurons were immunoassayed with anti‐MAP2 antibody and quantified for neurite degeneration ( n = 15 images per condition with N = 3 independent experiments). (C) cis ‐RSV and trans ‐RSV have opposite effects on nMAβ42‐mediated DNA damage accumulation. Immunostaining images (scale bar, 10 μm) for DNA damage marker, pSer139‐H2AX foci (γ‐H2AX, green; DAPI—nuclear marker, blue) in cortical neurons (DIV 10) treated with cis ‐RSV and trans ‐RSV (50 μM) alone or in combination with nMAβ 42 (50 nM) for 24 h. The graph represents the average number of γ‐H2AX foci per nuclei for n = 30 neurons per treatment condition for n = 3 independent experiments. (D) cis‐ RSV and trans ‐RSV have opposite effects on nMAβ42‐induced histone H3 phosphorylation. Immunostaining images (scale bar, 10 μm) for pSer‐10‐H3 (pSer‐10‐H3, green; DAPI—nuclear marker, blue) in cortical neurons (DIV 10) treated with cis ‐RSV and trans ‐RSV (50 μM) alone or in combination with nMAβ 42 (50 nM) for 24 h. The graph represents pSer‐10‐H3 for n = 30 neurons per condition for N = 3 independent experiments with p values represented in the figure (* ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001, and **** ≤ 0.0001). Statistical analysis was done using 2 way ANOVA with Tukey's multiple comparisons test.

Article Snippet: MAP2 , Aves labs , SKU: MAP , 1:500.

Techniques: Staining, Marker, Immunostaining, Phospho-proteomics