@article{c8d724b63f8947a8b97527a3ad2c64e5,
title = "A Viral Toolbox of Genetically Encoded Fluorescent Synaptic Tags",
abstract = "Fibronectin intrabodies generated with mRNA display (FingRs) are a recently developed tool for labeling excitatory or inhibitory synapses, with the benefit of not altering endogenous synaptic protein expression levels or synaptic transmission. Here, we generated a viral vector FingR toolbox that allows for multi-color, neuron-type-specific labeling of excitatory or inhibitory synapses in multiple brain regions. We screened various fluorophores, FingR fusion configurations, and transcriptional control regulations in adeno-associated virus (AAV) and retrovirus vector designs. We report the development of a red FingR variant and demonstrated dual labeling of excitatory and inhibitory synapses in the same cells. Furthermore, we developed cre-inducible FingR AAV variants and demonstrated their utility, finding that the density of inhibitory synapses in aspiny striatal cholinergic interneurons remained unchanged in response to dopamine depletion. Finally, we generated FingR retroviral vectors, which enabled us to track the development of excitatory and inhibitory synapses in hippocampal adult-born granule cells.",
keywords = "Biological Sciences, Neuroscience, Techniques in Neuroscience",
author = "Seth Bensussen and Sneha Shankar and Ching, {Kimberley H.} and Dana Zemel and Ta, {Tina L.} and Mount, {Rebecca A.} and Shroff, {Sanaya N.} and Gritton, {Howard J.} and Pierre Fabris and Hannah Vanbenschoten and Connor Beck and Man, {Heng Ye} and Xue Han",
note = "Funding Information: We thank J.P. Gilbert, Yuda Huo, and Margaret O'Connor for providing neuron cultures. We also thank the Boston University Micro and Nano Imaging Facility for providing access to the confocal microscope (supported by NIH S10OD024993). X.H. acknowledges funding from the NIH Director's Office (1DP2NS082126), NINDS (1R01NS109794, R21MH109941, R01NS087950), and NSF CBET-1848029. S.B. S.N.S. and R.A.M. acknowledge funding from the NIH/NIGMS T32 Quantitative Biology and Physiology Fellowship (GM008764) through the Boston University Biomedical Engineering Department. S.N.S acknowledges NIH F31 NS115421. S.S. H.V. and C.B. acknowledge funding support from the Boston University UROP program. S.B. and X.H. conceived of and designed all experiments and wrote the manuscript. X.H. supervised the study. S.B. S.S. H.V. S.N.S. R.A.M. and C.B. performed all molecular cloning. S.B. with help from H.V. packaged the viruses. S.B. T.L.T. and K.H.C. performed animal surgeries. H.J.G. provided transgenic animals and comments on experimental design. S.B. S.S. H.J.G. and T.L.T. performed immunohistology. S.B. S.S. and T.L.T. performed confocal imaging. S.B. S.S. with help from D.Z. P.F. and K.H.C. performed data analysis. H.-Y.M provided neuron cultures and comments on experimental design. All authors helped edit the manuscript. The authors declare no competing financial interests. Funding Information: We thank J.P. Gilbert, Yuda Huo, and Margaret O'Connor for providing neuron cultures. We also thank the Boston University Micro and Nano Imaging Facility for providing access to the confocal microscope (supported by NIH S10OD024993 ). X.H. acknowledges funding from the NIH Director's Office ( 1DP2NS082126 ), NINDS ( 1R01NS109794 , R21MH109941 , R01NS087950 ), and NSF CBET- 1848029 . S.B., S.N.S., and R.A.M. acknowledge funding from the NIH/ NIGMS T32 Quantitative Biology and Physiology Fellowship ( GM008764 ) through the Boston University Biomedical Engineering Department . S.N.S acknowledges NIH F31 NS115421 . S.S., H.V., and C.B. acknowledge funding support from the Boston University UROP program. Publisher Copyright: {\textcopyright} 2020 The Author(s)",
year = "2020",
month = jul,
day = "24",
doi = "10.1016/j.isci.2020.101330",
language = "English (US)",
volume = "23",
journal = "iScience",
issn = "2589-0042",
publisher = "Elsevier Inc.",
number = "7",
}