Peikon Ian D, Kebschull Justus M, Vagin Vasily V, Ravens Diana I, Sun Yu-Chi, Brouzes Eric, Corrêa Ivan R, Bressan Dario, Zador Anthony M
Watson School of Biological Sciences, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Nucleic Acids Res. 2017 Jul 7;45(12):e115. doi: 10.1093/nar/gkx292.
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.
神经回路的功能由其突触连接的细节决定。目前，唯一能够以单突触精度确定神经接线图（即“连接组”）的可用方法是基于成像方法，这些方法速度慢、劳动强度大且成本高。在此，我们提出了SYNseq，一种将连接组转化为能够利用现代高通量DNA测序的速度和低成本的形式的方法。在SYNseq中，每个神经元都用一个独特的随机核苷酸序列（一种RNA“条形码”）进行标记，该序列通过工程蛋白靶向到突触。然后，突触前和突触后神经元中的条形码通过跨越突触的蛋白质-蛋白质交联关联起来，从组织中提取出来，并连接成适合测序的形式。尽管我们未能开发出一种高效的条形码连接方案，妨碍了该方法的广泛应用，但我们预计随着进一步发展，SYNseq将能够以高速和低成本追踪复杂回路。
