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. 2019 Jul 12;13:322. doi: 10.3389/fncel.2019.00322

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Copyright © 2019 Penserga, Kudumala, Poulos and Godenschwege.

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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Drosophila Amyloid Precursor Protein (APP) is retrogradely transported and co-traffics with Neuroglian. (A) Diagram of Giant Fiber (GF) circuit in the Drosophila nervous system. The GFs synapse with the TTMn and the peripherally synapsing interneurons (PSI) in the ventral nerve cord (VNC). The circuitry output is via the innervation of the jump (TTM) and flight (DLM, dorsal longitudinal muscle) muscles by the TTMn and DLMn (dorsal longitudinal) motoneurons. Live-imaging of GF axons were performed in the CvC (blue square). Sites of dye injections for GF anatomy visualization and placement of stimulation and recording electrodes for electrophysiological analysis are indicated. Gray box shows area of morphological analysis of GF terminals by confocal microscopy in the ventral nerve cord (VNC). (B) Immunostaining of endogenous APPL (green top panel and white bottom panel) with an antibody against the N-terminal domain in wild type GFs and in GFs in which Lis1 was cell-autonomously knocked down. GFs (magenta, top panel) were labeled by dye injections of rhodamine-dextran into the GF axons. For better visualization, the GFs were digitally traced and 3D-reconstructed to extract anti-APPL labeling that localizes to the reconstructed GFs (bottom panel). APPL accumulation at GF-TTMn and GF-PSI contact sites in Lis1 knock down animals are indicated by arrows. Scale bar represents 30 μm. (C) Immunolabeling of GFP- tagged Nrg (green) and V5-tagged APPL (magenta) in adult GF axons co-expressed using the R68A06 Gal4-driver. Single confocal slices of the different channels in the same plane are shown separately and together to visualize co-localization (arrows). Scale bar represents 5 μm. (D) Live-imaging of co-expressed mCherry-tagged Nrg and GFP-tagged APPL in the GF axons (Supplementary Video S1). A small section of the GF axons was photobleached prior to acquisition to reduce background from Nrg labeling at the axonal membrane. Video was obtained at 1 frame per second. Kymographs of APPL-GFP and Nrg-mCherry alone as well as together are shown. Overlapping trajectory of APPL-GFP and Nrg-mCherry vesicles in retrograde direction are indicated by white arrows.