Volume 22, Issue 7, Pages (April 2012)

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Volume 22, Issue 7, Pages 590-595 (April 2012) A Novel Drosophila Model of Nerve Injury Reveals an Essential Role of Nmnat in Maintaining Axonal Integrity  Yanshan Fang, Lorena Soares, Xiuyin Teng, Melissa Geary, Nancy M. Bonini  Current Biology  Volume 22, Issue 7, Pages 590-595 (April 2012) DOI: 10.1016/j.cub.2012.01.065 Copyright © 2012 Elsevier Ltd Terms and Conditions

Current Biology 2012 22, 590-595DOI: (10.1016/j.cub.2012.01.065) Copyright © 2012 Elsevier Ltd Terms and Conditions

Figure 1 The Drosophila Wing Model of Axon Injury and Degeneration (A) The nerve tract along the wing margin (L1 and costal veins) is highlighted by mGFP using a dpr-GAL4 driver; asterisk, the last cluster of L1 neurons. (B) A higher magnification of the red box area is shown. Arrowheads, axons; arrow, dendrites. (C and D) On D10 after wing cut, axonal mGFP was undetectable in the wing arch (C) and the thoracic ganglion (D) of wild-type (WT) flies but still robust in WldS flies. Scale bar represents 100 μm in (A) and (C), 10 μm in (B), and 50 μm in (D). Current Biology 2012 22, 590-595DOI: (10.1016/j.cub.2012.01.065) Copyright © 2012 Elsevier Ltd Terms and Conditions

Figure 2 Time Course of Injury-Induced Axon Degeneration in the Wing Cut Model (A) Axon degeneration scores (see Figure S2B) of flies expressing UAS-Luciferase (Ctrl) or WldS, mNmnats, or dNmnat at indicated time after injury. Mean ± SEM is shown, n = 22∼35. (B) The thoracic projection was examined at indicated time after the right wing was cut. The left wing was uncut, and its projection was used as an internal control. Scale bar represents 20 μm. Current Biology 2012 22, 590-595DOI: (10.1016/j.cub.2012.01.065) Copyright © 2012 Elsevier Ltd Terms and Conditions

Figure 3 Knockdown of Nmnat or Milton in the Wing Nerve Induces Spontaneous Axon Degeneration (A and B) dpr>RNAi-dNmnat flies exhibited retrograde, spontaneous axon degeneration. Fragmentation of axonal mChRFP was seen earlier and more prominent in the wing arch (distal axons) than in the wing tip (proximal axons). (C) dpr>RNAi-Milton flies had normally distributed mitoGFP and smooth axonal mChRFP on D0. On D3, mitoGFP was depleted from the axons (wing arch) and retained in the cell bodies (wing tip); axonal mChRFP was massively fragmented in the wing arch but remained continuous in the wing tip. (D and E) Degeneration curves of dpr>RNAi-dNmnat (D) and dpr>RNAi-Milton (E) flies, plotted as proportion of fragmented wing nerve (see the scale in A). Ctrl, UAS-Luciferase. Mean ± SEM is shown, n = 14∼42. Scale bar represents 100 μm in (A), 10 μm in (B), and 20 μm in (C). Current Biology 2012 22, 590-595DOI: (10.1016/j.cub.2012.01.065) Copyright © 2012 Elsevier Ltd Terms and Conditions

Figure 4 Mitochondria Are Rapidly Depleted upon Nerve Injury or Knockdown of dNmnat (A) Injury induced rapid mitoGFP loss and mChRFP fragmentation in the severed wing nerve, which was dramatically preserved by upregulation of dNmnat. (B) dpr>RNAi-dNmnat flies had normal mitoGFP and mChRFP appearance on D0. On D3, mitoGFP became nearly undetectable and axonal mChRFP was massively fragmented. (C and D) Quantification of mitoGFP and mChRFP intensity in (A) and (B), respectively (also see Figure S4A). Mean ± SEM is shown, n = 6∼9. ∗p < 0.01, ∗∗∗p < 0.0001. Scale bar represents 20 μm. Current Biology 2012 22, 590-595DOI: (10.1016/j.cub.2012.01.065) Copyright © 2012 Elsevier Ltd Terms and Conditions