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. 2022 Apr 26;16:846425. doi: 10.3389/fnins.2022.846425

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Copyright © 2022 Mallik and Frank.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Rotenone affects neuromuscular junction (NMJ) morphology. (A–F) Representative confocal images of NMJ synapses at muscle 6/7 of (A–C) DMSO control, (D–F) Larvae fed with 25μM rotenone double immunolabeled with DLG (magenta) and HRP (green) antibodies. The NMJ morphological defects were observed in the indicated genotypes. The flies were reared at 25°C to conduct NMJ analysis. Scale bar: 10 μm. (G–J) Histograms showing the average total number of boutons, relative DLG area, bouton area and DLG area at muscle 6/7 of A2 hemisegment in (G) DMSO control (65.25 ± 3.976, N = 8), Larvae fed with 25μM rotenone (52.75 ± 3.514, N = 8), (H) DMSO control (6.742 ± 0.6156, N = 30), Larvae fed with 25μM rotenone (4.101 ± 0.3706, N = 30), (I) DMSO control (16.67 ± 1.461, N = 30), Larvae fed with 25μM rotenone (17.11 ± 0.8969, N = 30) and (J) DMSO control (23.41 ± 1.473, N = 30), Larvae fed with 25μM rotenone (21.21 ± 0.9790, N = 30) samples. Rotenone-treated larvae showed reduced boutons number and DLG area at the NMJs. Minimum 35 type-1b boutons from 4 animals (8 muscle 6/7 NMJs of A2 hemisegment) were used for quantification. p-values are indicated in the figure. Statistical analysis based on Student’s t-test for pairwise comparison. Error bars represent mean ± s.e.m.