Using Brainbow and GRASP for detailed reconstruction of complete circuits with light microscopy
Acquiring detailed and complete structure of neural circuits is one of the central problems of neuroscience. Until present, the only method for generation of such data was thought to be the circuit reconstruction from serial Electron Microscopy, which could require decades to complete a small circuit. Here, we suggest that already existing light microscopy and genetic tools may be used to obtain much faster and cheaper reconstructions of larger complete neural circuits. We describe how a collection of specifically prepared genetically targeted light probes of connectivity may be produced and used to systematically deduce feasible circuit configurations up to the point of pinpointing circuit’s structure exactly. We suggest to represent different such probes mathematically as constraints on the circuit’s connectivity matrix. This then leads to a prescription for how to combine results of such probes, performed in the same or different animals, computationally to obtain detailed structural information about the neural circuit. This paradigm may be employed with light probes of connectivity, such as recently described ChR2-assisted circuit mapping, GRASP or modified transsynaptic viruses, and gene expression techniques such as Brainbow, MARCM, or UAS/Gal4. It may be used in model organisms such as C. Elegans, Drosophila, zerbafish, mouse, etc. We exemplify this paradigm by showing how GRASP and Brainbow may be used for detailed reconstruction of complete neural circuit in C. Elegans within one week or less with essentially off-the-shelf genetic and light microscopy tools. Even though wiring diagram in C. Elegans is thought to be known from previous sEM reconstruction, unprecedented ability to extract such circuits routinely and rapidly in different settings will offer unique opportunities for systems neuroscience.
Conference:
Computational and systems
neuroscience 2009, Salt Lake City, UT, United States, 26 Feb - 3 Mar, 2009.
Presentation Type:
Poster and Short Oral Presentation
Topic:
Poster and Short Oral Presentations
Citation:
(2009). Using Brainbow and GRASP for detailed reconstruction of complete circuits with light microscopy.
Front. Syst. Neurosci.
Conference Abstract:
Computational and systems
neuroscience 2009.
doi: 10.3389/conf.neuro.06.2009.03.011
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Received:
29 Jan 2009;
Published Online:
29 Jan 2009.