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. 2022 Nov 4;13:6622. doi: 10.1038/s41467-022-34445-9

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a, b, c Fibrillar flight muscles (IFMs) stained for F - actin and mitochondria expressing mito-gfp driven by DMef2-Gal4 showing parallel aligned mitochondria that are large, tube-like, and packed between myofibrils (Scale Bars: 5 μm). d, e Representative 3D rendering of electron microscopic images of mitochondrial arrangement (yellow) and ER (magenta) in flight muscles. f, g, h Tubular jump muscles show mitochondria that are thin and elongated arranged in parallel mitochondrial networks (Scale Bars: 5 μm). i, j Representative 3D rendering of mitochondrial networks (yellow) and ER (magenta) in jump muscles. k In walking (leg) muscles, DMef2-Gal4 driven mito-gfp shows both parallel and grid-like mitochondrial networks (marked with dashed line, scale bar: 20 μm). l, m Tubular leg muscle Fiber I showing primarily parallel mitochondrial networks. n, o Tubular leg muscle Fiber II showing a grid-like mitochondrial network. p, q Tubular leg muscle Fiber III showing grid-like mitochondria. (Scale Bars: 5 μm for all). r, s Representative 3D rendering of mitochondrial network organization (yellow) and ER (magenta) in leg muscles. t Mitochondrial volume as a percent of total muscle volume (Flight muscles (IFM) (UAS-mito-gfp;Dmef2-Gal4), n = 5 animals; Jump muscles (TDT) (UAS-mito-gfp;Dmef2-Gal4), n = 5 animals; Leg Fiber I, n = 7 animals; Leg Fiber II, n = 9 animals; Leg Fiber III, n = 5 animals). u Quantification of mitochondrial network orientation. Dotted line represents parallel equal to perpendicular (IFM, n = 11 animals; Jump muscles (TDT), n = 12 animals; Leg Fiber I, n = 10 animals; Leg Fiber II, n = 9 animals; Leg Fiber III, n = 6 animals). v Endoplasmic reticulum (ER) volume as a percent of total muscle volume (IFM, n = 5 animals; Jump muscles (TDT), n = 5 animals; Leg Fiber I, n = 3 animals; Leg Fiber II, n = 4 animals; Leg Fiber III, n = 5 animals). Each point represents value for each animal dataset. Bars represent mean ± SD. Significance determined as p < 0.05 from one way ANOVA with Tukey’s (*, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001; ns, non-significant).