Using high-throughput barcode sequencing to efficiently map connectomes

Peikon, I. D., Kebschull, J. M., Vagin, V. V., Ravens, D. I., Sun, Y. C., Brouzes, E., Correa, I. R., Bressan, D., Zador, A. M. (2017) Using high-throughput barcode sequencing to efficiently map connectomes. Nucleic Acids Res. ISSN 0305-1048

URL: https://www.ncbi.nlm.nih.gov/pubmed/28449067
DOI: 10.1093/nar/gkx292

Abstract

The function of a neural circuit is determined by the details of its synaptic connections. At present, the only available method for determining a neural wiring diagram with single synapse precision-a 'connectome'-is based on imaging methods that are slow, labor-intensive and expensive. Here, we present SYNseq, a method for converting the connectome into a form that can exploit the speed and low cost of modern high-throughput DNA sequencing. In SYNseq, each neuron is labeled with a unique random nucleotide sequence-an RNA 'barcode'-which is targeted to the synapse using engineered proteins. Barcodes in pre- and postsynaptic neurons are then associated through protein-protein crosslinking across the synapse, extracted from the tissue, and joined into a form suitable for sequencing. Although our failure to develop an efficient barcode joining scheme precludes the widespread application of this approach, we expect that with further development SYNseq will enable tracing of complex circuits at high speed and low cost.

Item Type: Paper
Subjects: organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits

Investigative techniques and equipment > Whole Brain Circuit Mapping
CSHL Authors:
Communities: CSHL labs > Zador lab
Watson School > Publications
Depositing User: Matt Covey
Date Deposited: 28 Apr 2017 16:30
Last Modified: 28 Apr 2017 16:30
Related URLs:
URI: http://repository.cshl.edu/id/eprint/34646

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