A New Fiji-Based Algorithm That Systematically Quantifies Nine Synaptic Parameters Provides Insights into Drosophila NMJ Morphometry
Fig 3
Evaluation of reproducibility.
Macro assessment and quantification of NMJs on muscle 4. Ankyrin2 knockdown (w;UAS-Dicer-2/UAS-KK107238;elav-Gal4/+, n = 18 and w;UAS-Dicer-2/UAS-KK107369;elav-Gal4/+, n = 19) was confirmed to result in smaller NMJ size compared to genetic background controls (w;UAS-Dicer-2/+;elav-Gal4/+, n = 500) when using the macro (A-B). The macro-annotated outline is indicated in yellow. Highwire knockdown (w;UAS-Dicer-2/UAS-GD28163;elav-Gal4/+, n = 15 and w;UAS-Dicer-2/UAS-GD36085;elav-Gal4/+, n = 26) resulted in longer and more branched NMJs compared to genetic background control NMJs (w;UAS-Dicer-2/+;elav-Gal4/+, n = 402) (C-D). The macro-annotated skeleton is indicated in blue. Rab3 knockdown (w;UAS-Dicer-2/UAS-KK100787;elav-Gal4/+, n = 18) resulted in NMJs with a lower number of Brp-positive active zones compared to genetic background controls (w;UAS-Dicer-2/+;elav-Gal4/+, n = 476) (E-F). The macro-annotated Brp-positive active zones are indicated as white foci. Error bars indicate 95% confidence interval and asterisks represent significance (padj < 0.05).