Long-term labeling and imaging of synaptically-connected neuronal networks in vivo using nontoxic, double-deletion-mutant rabies viruses

Jin, L.; Sullivan, H. A.; Zhu, M.; Lavin, T. K.; Matsuyama, M.; Lea, N. E.; Xu, R.; Hou, Y.; Rutigliani, L.; Pruner, M. T.; Babcock, K. R.; Ip, J. P. K.; Hu, M.; Daigle, T. L.; Zeng, H.; Sur, M.; Wickersham, I. R.

doi:10.1101/2021.12.04.471186

bioRxiv · 10.1101/2021.12.04.471186

Long-term labeling and imaging of synaptically-connected neuronal networks in vivo using nontoxic, double-deletion-mutant rabies viruses

Jin, L.·Sullivan, H. A.·Zhu, M.·Lavin, T. K.·Matsuyama, M.·Lea, N. E.·Xu, R.·Hou, Y.·Rutigliani, L.·Pruner, M. T.·Babcock, K. R.·Ip, J. P. K.·Hu, M.·Daigle, T. L.·Zeng, H.·Sur, M.·Wickersham, I. R.

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Abstract

Monosynaptic tracing is a widely-used technique for mapping neural circuitry, but its cytotoxicity has confined it primarily to anatomical applications. Here we present a second-generation system for labeling direct inputs to targeted neuronal populations with minimal toxicity, using double-deletion-mutant rabies viruses. Spread of the viruses requires expression of both deleted viral genes in trans in postsynaptic source cells; suppressing this expression with doxycycline following an initial period of viral replication reduces toxicity to postsynaptic cells. Longitudinal two-photon imaging in vivo indicated that over 90% of both presynaptic and source cells survived for the full twelve-week course of imaging. Ex vivo whole-cell recordings at 5 weeks postinfection showed that the second-generation system perturbs input and source cells much less than does the first-generation system. Finally, two-photon calcium imaging of labeled networks of visual cortex neurons showed that their visual response properties appeared normal for 10 weeks, the longest we followed them.

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Jin, L., Sullivan, H. A., Zhu, M., Lavin, T. K., Matsuyama, M., Lea, N. E., Xu, R., Hou, Y., Rutigliani, L., Pruner, M. T., Babcock, K. R., Ip, J. P. K., Hu, M., Daigle, T. L., Zeng, H., Sur, M., Wickersham, I. R.. 2021-12-04. Long-term labeling and imaging of synaptically-connected neuronal networks in vivo using nontoxic, double-deletion-mutant rabies viruses. https://doi.org/10.1101/2021.12.04.471186

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