PUBLICATION
Visualizing neurons one-by-one in vivo: optical dissection and reconstruction of neural networks with reversible fluorescent proteins
- Authors
- Aramaki, S., and Hatta, K.
- ID
- ZDB-PUB-060921-9
- Date
- 2006
- Source
- Developmental Dynamics : an official publication of the American Association of Anatomists 235(8): 2192-2199 (Journal)
- Registered Authors
- Hatta, Kohei
- Keywords
- zebrafish, Dronpa, imaging, two-photon microscopy, neuron, neural network, confocal microscopy, Mauthner cell
- MeSH Terms
-
- Animals
- Brain/embryology
- Brain/metabolism
- Genes, Reporter/genetics
- Luminescent Proteins/analysis*
- Luminescent Proteins/genetics
- Luminescent Proteins/metabolism*
- Microscopy, Confocal/methods*
- Nerve Net/cytology*
- Nerve Net/embryology
- Nerve Net/metabolism*
- Neurons/metabolism*
- Optics and Photonics
- Time Factors
- Zebrafish
- PubMed
- 16607643 Full text @ Dev. Dyn.
Citation
Aramaki, S., and Hatta, K. (2006) Visualizing neurons one-by-one in vivo: optical dissection and reconstruction of neural networks with reversible fluorescent proteins. Developmental Dynamics : an official publication of the American Association of Anatomists. 235(8):2192-2199.
Abstract
A great many axons and dendrites intermingle to fasciculate, creating synapses as well as glomeruli. During live imaging in particular, it is often impossible to distinguish between individual neurons when they are contiguous spatially and labeled in the same fluorescent color. In an attempt to solve this problem, we have taken advantage of Dronpa, a green fluorescent protein whose fluorescence can be erased with strong blue light, and reversibly highlighted with violet or ultraviolet light. We first visualized a neural network with fluorescent Dronpa using the Gal4-UAS system. During the time-lapse imaging of axonal navigation, we erased the Dronpa fluorescence entirely; re-highlighted it in a single neuron anterogradely from the soma or retrogradely from the axon; then repeated this procedure for other single neurons. After collecting images of several individual neurons, we then recombined them in multiple pseudo-colors to reconstruct the network. We have also successfully re-highlighted Dronpa using two-photon excitation microscopy to label individual cells located inside of tissues and were able to demonstrate visualization of a Mauthner neuron extending an axon. These "optical dissection" techniques have the potential to be automated in the future and may provide an effective means to identify gene function in morphogenesis and network formation at the single cell level.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping