Journal: Frontiers in Pharmacology
Article Title: Chronic nicotine exposure induces molecular and transcriptomic endophenotypes associated with mood and anxiety disorders in a cerebral organoid neurodevelopmental model
doi: 10.3389/fphar.2024.1473213
Figure Lengend Snippet: Nicotine selectively upregulates certain nAchR populations at D42. (A) Immunofluorescent images captured using confocal microscopy of α 7 (green), α 4 (magenta) and β 2 nAchR (red) in brain organoids treated with (0.1, 1 or 10 μM) nicotine or without (VEH) for 14 days. Scale bar = 50 μm. ( B–D) Quantification of immunofluorescent images by the number of particles per area (mm 3 ). Nicotine did not affect α 7 nAchR expression (B) but significantly increased α 4 (C) and β 2 (D) compared to VEH organoids. Comparisons were made with one-way ANOVA or Kruskal Wallis followed by Fisher’s LSD post hoc test. Data are mean ± SEM, n = 3 organoids per group; 18–20 total ROIs per marker, *** p < 0.001, * p < 0.05, ns = not significant, p > 0.05. Each data point represents one ROI.
Article Snippet: Primary antibodies included ⍺ 7 nAchR (rabbit, Alomone labs ANC-007, 1:50), ⍺ 4 nAchR (mouse, Santa Cruz sc-74519, 1:50), β 2 nAchR (goat, Abcam ab189174, 1:100), D1R (rabbit, Abcam ab40653, 1:50), D2R (mouse, Sigma-Aldrich MABN53, 1:200), mGLUR2/3 (rabbit, Sigma-Aldrich 06–676, 1:50), GAD67 (mouse, Sigma-Aldrich mab5406, 1:50), Ki67 (rabbit, Abcam ab15581, 1:500), PROX1 (mouse, Sigma-Aldrich mab5654, 1:100), FZD9 (goat, Abcam ab110886, 1:100), FGFR1 (rabbit, Abcam ab0646, 1:200), CCasp3 (rabbit, Cell signaling 9661, 1:200), CTIP2 (rat, abcam ab18465, 1:100), CDH13 (goat, Novus Biologicals, AF3264, 1:200), MAP2 (mouse, Sigma-Aldrich mab3418, 1:100), NR2B (goat, Novus Biologicals NB100-41097, 1:50), GAT-1 (rabbit, Rockland Immunochemicals 612-401-D56, 1:100) and PV (mouse, Sigma-Aldrich P3088, 1:50).
Techniques: Confocal Microscopy, Expressing, Marker
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