Cited 2 time in webofscience Cited 2 time in scopus

Labeling Dual Presynaptic Inputs using cFork Anterograde Tracing System

Title
Labeling Dual Presynaptic Inputs using cFork Anterograde Tracing System
Authors
Oh, Jun-YoungHan, Jeong-HoLee, HyoeunHan, Young-EunRah, Jong CheolPark, Hyungju
DGIST Authors
Oh, Jun-Young; Han, Jeong-Ho; Lee, Hyoeun; Han, Young-Eun; Rah, Jong Cheol; Park, Hyungju
Issue Date
2020-06
Citation
Experimental Neurobiology, 29(3), 219-229
Type
Article
Article Type
Article
Author Keywords
EGFPmScarletAnterograde tracingAdeno-associated virus -1Adeno-associated virus -5Brain mapping
Keywords
MOTOR CORTEXNEURONSORGANIZATIONPROJECTIONSPATHWAYSFIELD
ISSN
1226-2560
Abstract
Understanding brain function-related neural circuit connectivity is essential for investigating how cognitive functions are decoded in neural circuits. Trans-synaptic viral vectors are useful for identifying neural synaptic connectivity because of their ability to be transferred from transduced cells to synaptically connected cells. However, concurrent labeling of multisynaptic inputs to postsynaptic neurons is impossible with currently available trans-synaptic viral vectors. Here, we report a neural circuit tracing system that can simultaneously label postsynaptic neurons with two different markers, the expression of which is defined by presynaptic input connectivity. This system, called “cFork (see fork)”, includes delivering serotype 1-packaged AAV vectors (AAV1s) containing Cre or flippase recombinase (FlpO) into two different presynaptic brain areas, and AAV5 with a dual gene expression cassette in postsynaptic neurons. Our in vitro and in vivo tests showed that selective expression of two different fluorescence proteins, EGFP and mScarlet, in postsynaptic neurons could be achieved by AAV1-mediated anterograde trans-synaptic transfer of Cre or FlpO constructs. When this tracing system was applied to the somatosensory barrel field cortex (S1BF) or striatum innervated by multiple presynaptic inputs, postsynaptic neurons defined by presynaptic inputs were simultaneously labeled with EGFP or mScarlet. Our new anterograde tracing tool may be useful for elucidating the complex multisynaptic connectivity of postsynaptic neurons regulating diverse brain functions. Copyright © Experimental Neurobiology 2020.
URI
http://hdl.handle.net/20.500.11750/12847
DOI
10.5607/en20006
Publisher
한국뇌신경과학회
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