Monosynaptic restriction of the anterograde herpes simplex virus strain H129 for neural circuit tracing

Kyle B. Fischer, Hannah K. Collins, Yan Pang, Dheeraj S. Roy, Ying Zhang, Guoping Feng, Shu Jing Li, Adam Kepecs, Edward M. Callaway

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Identification of synaptic partners is a fundamental task for systems neuroscience. To date, few reliable techniques exist for whole brain labeling of downstream synaptic partners in a cell-type-dependent and monosynaptic manner. Herein, we describe a novel monosynaptic anterograde tracing system based on the deletion of the gene UL6 from the genome of a cre-dependent version of the anterograde Herpes Simplex Virus 1 strain H129. Given that this knockout blocks viral genome packaging and thus viral spread, we reasoned that co-infection of a HSV H129 ΔUL6 virus with a recombinant adeno-associated virus expressing UL6 in a cre-dependent manner would result in monosynaptic spread from target cre-expressing neuronal populations. Application of this system to five nonreciprocal neural circuits resulted in labeling of neurons in expected projection areas. While some caveats may preclude certain applications, this system provides a reliable method to label postsynaptic partners in a brain-wide fashion.

Original languageEnglish
Pages (from-to)584-595
Number of pages12
JournalJournal of Comparative Neurology
Issue number5
StatePublished - Apr 2023


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