A genetically encoded tool for reconstituting synthetic modulatory neurotransmission and reconnect neural circuits in vivo

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Hawk, J. D., Wisdom, E. M., Sengupta, T., Kashlan, Z. D., & Colon-Ramos, D. A. A genetically encoded tool for reconstituting synthetic modulatory neurotransmission and reconnect neural circuits in vivo. Nature Communications, 12(1), (2021): 4795, https://doi.org/10.1038/s41467-021-24690-9.Chemogenetic and optogenetic tools have transformed the field of neuroscience by facilitating the examination and manipulation of existing circuits. Yet, the field lacks tools that enable rational rewiring of circuits via the creation or modification of synaptic relationships. Here we report the development of HySyn, a system designed to reconnect neural circuits in vivo by reconstituting synthetic modulatory neurotransmission. We demonstrate that genetically targeted expression of the two HySyn components, a Hydra-derived neuropeptide and its receptor, creates de novo neuromodulatory transmission in a mammalian neuronal tissue culture model and functionally rewires a behavioral circuit in vivo in the nematode Caenorhabditis elegans. HySyn can interface with existing optogenetic, chemogenetic and pharmacological approaches to functionally probe synaptic transmission, dissect neuropeptide signaling, or achieve targeted modulation of specific neural circuits and behaviors.This work was initiated in the Grass Laboratory at the Marine Biological Laboratories (MBL) with funding through a Grass Fellowship awarded to J.D.H. Thanks to Richard Goodman (OHSU) for encouragement during the conceptualization of the fellowship application, and the 2019 Grass Fellows, Mel Coleman (Grass Director), and Christophe Dupré (Associate Director) for advice and support during the summer fellowship. We thank the MBL Division of Education and participants in the Vendor Equipment Loan Program. Special thanks to Sutter Instruments, who generously provided all electrophysiology equipment and substantial on-site assistance, and Zeiss, who provided on-site assistance at MBL. We thank Zhao-Wen Wang and Ping Liu (UConn) for guidance and training in patch-clamp electrophysiology, as well as providing Neuro2a cells. We thank Rob Steele (UCI) for supplying Hydra, as well as advice and inspiration on Hydra biology. We thank members of the Colón-Ramos lab and Hari Shroff (NIH) for thoughtful comments on the manuscript. We thank Michael Koelle and Andrew Olson (Yale University) for advice and reagents regarding serotonin rewiring experiments. We also thank Steve Flavell (MIT) for ideas and reagents regarding the experiments associated with del-7. We thank Life Science Editors for editing assistance. D.A.C.-R. is an MBL Fellow. Research in the D.A.C.-R. lab was supported by NIH R01NS076558, DP1NS111778, and by an HHMI Scholar Award