Table 1.
Fig 1.
Mio expression in muscle is necessary for normal glycogen storage.
(A) Analysis of Mio expression from thoraxes of 5–7 day old Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR flies compared to Mef2-Gal4>GFP controls (n = 8). Mio levels of the Mef2-Gal4>GFP controls were set to 1.0 and Mio levels of Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR animals were then calculated relative to their respective control. Values represent mean Mio expression±SEM. *p<0.05 by One-way ANOVA with post hoc Tukey test. (B) Glycogen/protein and (C) triglyceride/protein of thoraxes dissected from 5–8 day old Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR male and female flies compared to their respective Mef2-Gal4>GFP controls (n = 6). Values represent mean±SEM. *p<0.05 by One-way ANOVA with post hoc Tukey test.
Fig 2.
Mio in muscle is necessary for normal flight.
Flight tests were performed on Mef2-Gal4>MiodsRNA (n = 144) and Mef2-Gal4>Mio-IR (n = 296) flies and compared to Mef2-Gal4>GFP (n = 179) controls and scored based on flight ability (See methods). Values represent average flight score ±SEM. *p<0.05 Kruskal-Wallis One-way ANOVA with post hoc all pairwise multiple comparison pooled sample median test.
Fig 3.
Decreasing Mio levels results in abnormal myofibril ultrastructure.
Transmission Electron Microscopy of Indirect Flight Muscles of adult Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR females compared to Mef2-Gal4>GFP controls. Panels (A), (B) and (C) show cross sections of the myofibrils; panels (D), (E) and (F) show longitudinal sections of myofibrils. f, myofibril; c, mitochondrion; g, glycogen granules; m, m-line; z, z-line. Scale bar = 0.5μm.
Fig 4.
Scanning Electron Microscopy of Indirect Flight Muscles of adult Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR females compared to Mef2-Gal4>GFP controls. Panels (A), (B) and (C) show cross sections of the myofibrils; bars indicate 0.5μm. f, myofibril; c, mitochondrion; g, glycogen granules. (D) Average myofibril area of Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR flies compared to Mef2-Gal4>GFP control flies (n = 3–5). All values represent average myofibril area ±SEM. *p<0.05 Kruskal-Wallis One-way ANOVA with post hoc all pairwise multiple comparison pooled sample median test.
Fig 5.
Decreasing Mio levels results in abnormal myofibril organization.
Scanning Electron Microscopy of longitudinal sections of Indirect Flight Muscles of adult Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR females compared to Mef2-Gal4>GFP controls. f, myofibril; g, glycogen granules. Each arrow points to a single longitudinal myofibril. Scale bar = 20μm.
Fig 6.
Mio affects myofibril size in pharate adults.
Transmission Electron Microscopy of Indirect Flight Muscles of Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR pharate adults compared to Mef2-Gal4>GFP controls. Panels (A), (B) and (C) show cross sections of the myofibrils. Bars indicate 0.5μm. f, myofibril; c, mitochondrion; g, glycogen granules. (D) Average myofibril area of Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR pharate adults compared to Mef2-Gal4>GFP controls (n = 3–5). Values represent average myofibril area ±SEM. *p<0.05 by One-way ANOVA with post hoc Tukey test.
Fig 7.
Mio is not necessary for normal expression of structural genes in adult fly muscle.
Expression of Actin 88F (Act88F, n = 8), Myosin Heavy Chain (MHC, n = 11), and Myosin light chain-2 (Mlc-2, n = 8) was measured by performing quantitative PCR on thorax cDNA from 5–7 day old Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR flies and compared to Mef2-Gal4>GFP controls. mRNA levels of Mef2-Gal4>GFP controls were set to 1.0 and mRNA levels of Mef2-Gal4>MiodsRNA and Mef2-Gal4>Mio-IR animals were then normalized to their respective control. Values represent mean±SEM. *p<0.05 by One-way ANOVA with post hoc Tukey test.