Figure 8

Altered neuronal morphology in continuous ethanol exposure. (A–C). Neuronal morphology. (A) Comparison of changes in number of neurites in D8 control and continuous ethanol-treated cells. Representative images from D8 control (left panel), continuous ethanol exposure (right panel). Scale bars represent 20 μm. (B) Quantification of the number of neurites. From each biological replicate (N = 3), at least 20 TUB III⁺ neuronal cells per were quantified. N = 3 ± SEM. (C) Box and whisker plot of the length of the longest neurite in D8 control and continuous ethanol exposure groups. From each biological replicate (N = 3), at least 20 TUB III⁺ neuronal cells per were quantified. N = 3 ± SEM. Significant differences from controls are indicated with P < 0.05, **P < 0.01, ***P < 0.001, or ****P < 0.0001. (D-E) Glial morphology. (D) Representative images of GFAP⁺ D8 control astrocytes (left), and continuous ethanol exposed astrocytes (right). Scale bars represent 20 μm. Note that no significant change in nuclear size was observed between control and ethanol treatment. (E) Comparison of the range of glial cell sizes in (µm²) in D8 control and continuous ethanol-treated populations. At least 20 GFAP⁺ cells per biological replicate were quantified under each condition. N = 3 ± SEM. ns: not significant. (F) Gfap transcript expression values normalized to FPKM (Fragments per kilobase per million reads) in D8 control and continuous ethanol exposure. N = 3 ± SEM. Significant difference from control is indicated with ***P < 0.001. (G) Cellular assembly and organization IPA analysis showing the gene networks associated with misregulation of organization of cytoskeleton. “Data were analyzed through the use of IPA (QIAGEN Inc., https://www.qiagenbioinformatics.com/products/ingenuity-pathway-analysis).”