Journal: iScience
Article Title: Altering heparan sulfate suppresses cell abnormalities and neuron loss in Drosophila presenilin model of Alzheimer Disease
doi: 10.1016/j.isci.2024.110256
Figure Lengend Snippet: Reduction of sfl function reduces Psn -knockdown-induced neurodegenerative vacuole formation and apoptosis levels (A) Representative images of Drosophila brains stained with Alexa Fluor 594-Phalloidin and imaged by confocal microscopy. Top row panels show low magnification images of the entire brain; bottom row: higher magnification (see scale bars). White arrows indicate neurodegenerative vacuoles. Black arrows indicate flares showing the higher staining intensity of phalloidin associated with and near vacuoles. Black arrowheads indicate tracheoles, air conducting tubules, discernable in the serial confocal sections. (B) Bar graphs showing average vacuole area and count per brain in female Drosophila. Vacuole area and number of vacuoles per brain are significantly increased in shPsn3 animals compared to controls and significantly elevated compared to shPsn3;sfl RNAi animals 7 days post-eclosion (ANOVA Tukey test: w RNAi n = 10, shPsn3 n = 24, shPsn3;sfl RNAi n = 14, sfl RNAi = 20; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001); error bars represent standard error of the mean (SEM). Open symbols denote significance values for 7-day-old animals and closed symbols for 30-day-old animals. Number of symbols indicates levels of significance between the two groups with those symbols. Vacuole area and number of vacuoles per brain are significantly increased in shPsn3 animals 30 days post-eclosion compared to controls, and double shPsn3;sfl RNAi animals showed a significant reduction in vacuole number compared to shPsn3 . Vacuole area was reduced in shPsn3;sfl RNAi animals compared to shPsn3 but did not achieve significance (ANOVA Tukey test: w RNAi n = 13, shPsn3 n = 20, shPsn3;sfl RNAi n = 8, sfl RNAi = 20; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). (C) Representative images of Drosophila brains stained with the apoptosis marker, acridine orange. Micrographs are pixel maximum intensity stacks from confocal serial sections. (D) Bar graphs quantifying apoptotic staining in optic lobes of adult female Drosophila , age matched 1–3 weeks. Apoptosis is significantly higher in shPsn3 than controls ( w RNAi ) and significantly lower in shPsn3;sfl RNAi compared to shPsn3 animals (ANOVA Tukey test: w RNAi n = 10, shPsn3 n = 10, shPsn3;sfl RNAi n = 10, sfl RNAi n = 10; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). (E) Bar graphs demonstrating average performance on negative geotaxis assays in adult female Drosophila , aged 4–7 days. Performance is statistically significant between w RNAi and all other genotypes (ANOVA Tukey test: w RNAi n = 25, shPsn3 n = 20, shPsn3;sfl RNAi n = 25, sfl RNAi = 35; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). shPsn3;sfl RNAi show significantly improved geotaxis compared to shPsn3 animals ( p < 0.05).
Article Snippet: ANOVA statistical analysis (one-way, Tukey comparison) was performed in Minitab to confirm significance.
Techniques: Knockdown, Staining, Confocal Microscopy, Marker
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