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WTR: A Toolkit for Functional Anterograde Transsynaptic Circuit Mapping

Wang, T. A.; Chen, C.; Liu, R.; Yi-Luo, A.; Cao, X.; Hu, J.; Guan, S.; Chang, S.-y.; Cui, X.; Zhou, W.; Zhao, F.; Huang, C.-T.; Duan, X.; Jan, L. Y.

2026-03-25 neuroscience
10.64898/2026.03.23.713595 bioRxiv
Show abstract

The brain coordinates animal physiology and behavior via neuronal circuits. To understand and simulate brain functions, it is essential to delineate the synaptic connectivity between neurons. Transsynaptic tracers serve as powerful tools for such purposes. In response to the demand for anterograde tracers for circuit mapping and functional interrogation, we developed WTR, a fusion protein of mammalian codon-optimized WGA, TEV-protease cleavage sequence, and Recombinase. WTR expressed via AAV vectors in cell-type-specific starter neurons reaches their postsynaptic neurons and releases Cre/Flpo upon exposure to TEV-protease expressed in downstream neurons. Accompanied by Cre/Flpo-dependent expression of EGFP, GCaMP7s, or ChR2, the toolkit enables labeling, recording, or manipulation of downstream neurons. We utilized WTR to characterize downstream neurons of either glutamatergic or GABAergic neurons in the preoptic area of anterior hypothalamus for their differential actions in thermoregulation or stress responses, respectively. These results establish WTR as a versatile platform for functional anterograde circuit mapping.

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