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@ARTICLE{Sugie:139158,
      author       = {Sugie, Atsushi and Möhl, Christoph and Hakeda-Suzuki,
                      Satoko and Matsui, Hideaki and Suzuki, Takashi and
                      Tavosanis, Gaia},
      title        = {{A}nalyzing {S}ynaptic {M}odulation of {D}rosophila
                      melanogaster {P}hotoreceptors after {E}xposure to
                      {P}rolonged {L}ight.},
      journal      = {JoVE journal},
      volume       = {Neuroscience},
      number       = {120},
      issn         = {1940-087X},
      address      = {New Delhi},
      publisher    = {JoVE124831},
      reportid     = {DZNE-2020-05480},
      pages        = {55176},
      year         = {2017},
      abstract     = {The nervous system has the remarkable ability to adapt and
                      respond to various stimuli. This neural adjustment is
                      largely achieved through plasticity at the synaptic level.
                      The Active Zone (AZ) is the region at the presynaptic
                      membrane that mediates neurotransmitter release and is
                      composed of a dense collection of scaffold proteins. AZs of
                      Drosophila melanogaster (Drosophila) photoreceptors undergo
                      molecular remodeling after prolonged exposure to natural
                      ambient light. Thus the level of neuronal activity can
                      rearrange the molecular composition of the AZ and contribute
                      to the regulation of the functional output. Starting from
                      the light exposure set-up preparation to the
                      immunohistochemistry, this protocol details how to quantify
                      the number, the spatial distribution, and the delocalization
                      level of synaptic molecules at AZs in Drosophila
                      photoreceptors. Using image analysis software, clusters of
                      the GFP-fused AZ component Bruchpilot were identified for
                      each R8 photoreceptor (R8) axon terminal. Detected
                      Bruchpilot spots were automatically assigned to individual
                      R8 axons. To calculate the distribution of spot frequency
                      along the axon, we implemented a customized software plugin.
                      Each axon's start-point and end-point were manually defined
                      and the position of each Bruchpilot spot was projected onto
                      the connecting line between start and end-point. Besides the
                      number of Bruchpilot clusters, we also quantified the
                      delocalization level of Bruchpilot-GFP within the clusters.
                      These measurements reflect in detail the spatially resolved
                      synaptic dynamics in a single neuron under different
                      environmental conditions to stimuli.},
      keywords     = {Animals / Drosophila Proteins: metabolism / Drosophila
                      melanogaster: metabolism / Green Fluorescent Proteins:
                      metabolism / Light / Luminescent Agents: metabolism /
                      Photoreceptor Cells, Invertebrate: metabolism /
                      Photoreceptor Cells, Invertebrate: radiation effects /
                      Presynaptic Terminals / Protein Binding / Protein Transport
                      / Synapses: metabolism / Synaptic Transmission: physiology /
                      Drosophila Proteins (NLM Chemicals) / Luminescent Agents
                      (NLM Chemicals) / Green Fluorescent Proteins (NLM
                      Chemicals)},
      cin          = {AG Tavosanis / IDAF},
      ddc          = {570},
      cid          = {I:(DE-2719)1013018 / I:(DE-2719)1040200},
      pnm          = {342 - Disease Mechanisms and Model Systems (POF3-342)},
      pid          = {G:(DE-HGF)POF3-342},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:28287587},
      pmc          = {pmc:PMC5408834},
      doi          = {10.3791/55176},
      url          = {https://pub.dzne.de/record/139158},
}