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All-viral tracing of monosynaptic inputs to single birthdate-defined neurons in the intact brain
Journal
Cell reports methods
Volume
2
Number
5
Pages / Article-Number
100221
Keywords
AAV; calcium imaging; hippocampus; in vivo imaging; medial entorhinal cortex; monosynaptic tracing; optogenetics; rabies virus; single cell
Abstract
Neuronal firing patterns are the result of inputs converging onto single cells. Identifying these inputs, anatomically and functionally, is essential to understand how neurons integrate information. Single-cell electroporation of helper genes and subsequent local injection of recombinant rabies viruses enable precise mapping of inputs to individual cells in superficial layers of the intact cortex. However, access to neurons in deeper structures requires more invasive procedures, including removal of overlying tissue. We developed a method that, through a combination of virus injections, allows us to target 4 or fewer hippocampal cells 48% of the time and a single cell 16% of the time in wild-type mice without use of electroporation or tissue aspiration. We identify local and distant monosynaptic inputs that can be functionally characterized; in vivo; . By expanding the toolbox for monosynaptic circuit tracing, this method will help further our understanding of neuronal integration at the level of single cells.
