TY  - JOUR
AU  - Sugie, Atsushi
AU  - Möhl, Christoph
AU  - Hakeda-Suzuki, Satoko
AU  - Matsui, Hideaki
AU  - Suzuki, Takashi
AU  - Tavosanis, Gaia
TI  - Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light.
JO  - JoVE journal
VL  - Neuroscience
IS  - 120
SN  - 1940-087X
CY  - New Delhi
PB  - JoVE124831
M1  - DZNE-2020-05480
SP  - 55176
PY  - 2017
AB  - 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.
KW  - Animals
KW  - Drosophila Proteins: metabolism
KW  - Drosophila melanogaster: metabolism
KW  - Green Fluorescent Proteins: metabolism
KW  - Light
KW  - Luminescent Agents: metabolism
KW  - Photoreceptor Cells, Invertebrate: metabolism
KW  - Photoreceptor Cells, Invertebrate: radiation effects
KW  - Presynaptic Terminals
KW  - Protein Binding
KW  - Protein Transport
KW  - Synapses: metabolism
KW  - Synaptic Transmission: physiology
KW  - Drosophila Proteins (NLM Chemicals)
KW  - Luminescent Agents (NLM Chemicals)
KW  - Green Fluorescent Proteins (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:28287587
C2  - pmc:PMC5408834
DO  - DOI:10.3791/55176
UR  - https://pub.dzne.de/record/139158
ER  -