Back to Search
Start Over
Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light
- Source :
- JoVE journal Neuroscience(120), 55176 (2017). doi:10.3791/55176
- Publication Year :
- 2017
- Publisher :
- JoVE124831, 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.
- Subjects :
- Luminescent Agents
Light
metabolism [Luminescent Agents]
Green Fluorescent Proteins
Presynaptic Terminals
metabolism [Drosophila Proteins]
metabolism [Synapses]
Synaptic Transmission
metabolism [Photoreceptor Cells, Invertebrate]
Protein Transport
Drosophila melanogaster
physiology [Synaptic Transmission]
ddc:570
metabolism [Green Fluorescent Proteins]
Synapses
metabolism [Drosophila melanogaster]
Animals
radiation effects [Photoreceptor Cells, Invertebrate]
Drosophila Proteins
Photoreceptor Cells, Invertebrate
Neuroscience
Protein Binding
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- JoVE journal Neuroscience(120), 55176 (2017). doi:10.3791/55176
- Accession number :
- edsair.pmid.dedup....a475b61d5094406c99ed4553a7d377b7