Monosynaptic Tracing using Modified Rabies Virus Reveals Early and Extensive Circuit Integration of Human Embryonic Stem Cell-Derived Neurons
Shane Grealish, Andreas Heuer, Tiago Cardoso, Agnete Kirkeby, Marie Jönsson, Jenny Johansson, Anders Björklund, Johan Jakobsson, Malin Parmar, Shane Grealish, Andreas Heuer, Tiago Cardoso, Agnete Kirkeby, Marie Jönsson, Jenny Johansson, Anders Björklund, Johan Jakobsson, Malin Parmar
Abstract
Human embryonic stem cell (hESC)-derived dopamine neurons are currently moving toward clinical use for Parkinson's disease (PD). However, the timing and extent at which stem cell-derived neurons functionally integrate into existing host neural circuitry after transplantation remain largely unknown. In this study, we use modified rabies virus to trace afferent and efferent connectivity of transplanted hESC-derived neurons in a rat model of PD and report that grafted human neurons integrate into the host neural circuitry in an unexpectedly rapid and extensive manner. The pattern of connectivity resembled that of local endogenous neurons, while ectopic connections were not detected. Revealing circuit integration of human dopamine neurons substantiates their potential use in clinical trials. Additionally, our data present rabies-based tracing as a valuable and widely applicable tool for analyzing graft connectivity that can easily be adapted to analyze connectivity of a variety of different neuronal sources and subtypes in different disease models.
Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.
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References
- Barker R.A., Barrett J., Mason S.L., Björklund A. Fetal dopaminergic transplantation trials and the future of neural grafting in Parkinson’s disease. Lancet Neurol. 2013;12:84–91.
- Deshpande A., Bergami M., Ghanem A., Conzelmann K.K., Lepier A., Götz M., Berninger B. Retrograde monosynaptic tracing reveals the temporal evolution of inputs onto new neurons in the adult dentate gyrus and olfactory bulb. Proc. Natl. Acad. Sci. USA. 2013;110:E1152–E1161.
- Dunnett S.B., Nathwani F., Björklund A. The integration and function of striatal grafts. In: Björklund A., Dunnett S.B., editors. Progress in Brain Research. Elsevier; 2000. pp. 345–380.
- Etessami R., Conzelmann K.K., Fadai-Ghotbi B., Natelson B., Tsiang H., Ceccaldi P.E. Spread and pathogenic characteristics of a G-deficient rabies virus recombinant: an in vitro and in vivo study. J. Gen. Virol. 2000;81:2147–2153.
- Fisher L.J., Young S.J., Tepper J.M., Groves P.M., Gage F.H. Electrophysiological characteristics of cells within mesencephalon suspension grafts. Neuroscience. 1991;40:109–122.
- Gerfen C.R., Wilson C.J. The basal ganglia. In: Björklund A., Swanson L.W., Hökfelt T., editors. Handbook of Chemical Neuroanatomy. Elsevier; 1996. pp. 371–468.
- Grealish S., Diguet E., Kirkeby A., Mattsson B., Heuer A., Bramoulle Y., Van Camp N., Perrier A.L., Hantraye P., Björklund A., Parmar M. Human ESC-derived dopamine neurons show similar preclinical efficacy and potency to fetal neurons when grafted in a rat model of Parkinson’s disease. Cell Stem Cell. 2014;15:653–665.
- Kirkeby A., Grealish S., Wolf D.A., Nelander J., Wood J., Lundblad M., Lindvall O., Parmar M. Generation of regionally specified neural progenitors and functional neurons from human embryonic stem cells under defined conditions. Cell Rep. 2012;1:703–714.
- Kriks S., Shim J.W., Piao J., Ganat Y.M., Wakeman D.R., Xie Z., Carrillo-Reid L., Auyeung G., Antonacci C., Buch A. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease. Nature. 2011;480:547–551.
- Marshel J.H., Mori T., Nielsen K.J., Callaway E.M. Targeting single neuronal networks for gene expression and cell labeling in vivo. Neuron. 2010;67:562–574.
- Miyamichi K., Amat F., Moussavi F., Wang C., Wickersham I., Wall N.R., Taniguchi H., Tasic B., Huang Z.J., He Z. Cortical representations of olfactory input by trans-synaptic tracing. Nature. 2011;472:191–196.
- Osakada F., Callaway E.M. Design and generation of recombinant rabies virus vectors. Nat. Protoc. 2013;8:1583–1601.
- Sørensen A.T., Thompson L., Kirik D., Björklund A., Lindvall O., Kokaia M. Functional properties and synaptic integration of genetically labelled dopaminergic neurons in intrastriatal grafts. Eur. J. Neurosci. 2005;21:2793–2799.
- Thompson L., Barraud P., Andersson E., Kirik D., Bjorklund A. Identification of dopaminergic neurons of nigral and ventral tegmental area subtypes in grafts of fetal ventral mesencephalon based on cell morphology, protein expression, and efferent projections. J. Neurosci. 2005;25:6467–6477.
- Tønnesen J., Kokaia M. Electrophysiological investigations of synaptic connectivity between host and graft neurons. Prog. Brain Res. 2012;200:97–112.
- Tønnesen J., Parish C.L., Sørensen A.T., Andersson A., Lundberg C., Deisseroth K., Arenas E., Lindvall O., Kokaia M. Functional integration of grafted neural stem cell-derived dopaminergic neurons monitored by optogenetics in an in vitro Parkinson model. PLoS ONE. 2011;6:e17560.
- Ugolini G. Specificity of rabies virus as a transneuronal tracer of motor networks: transfer from hypoglossal motoneurons to connected second-order and higher order central nervous system cell groups. J. Comp. Neurol. 1995;356:457–480.
- Van der Werf Y.D., Witter M.P., Groenewegen H.J. The intralaminar and midline nuclei of the thalamus. Anatomical and functional evidence for participation in processes of arousal and awareness. Brain Res. Brain Res. Rev. 2002;39:107–140.
- Vivar C., Potter M.C., Choi J., Lee J.Y., Stringer T.P., Callaway E.M., Gage F.H., Suh H., van Praag H. Monosynaptic inputs to new neurons in the dentate gyrus. Nat. Commun. 2012;3:1107.
- Wall N.R., De La Parra M., Callaway E.M., Kreitzer A.C. Differential innervation of direct- and indirect-pathway striatal projection neurons. Neuron. 2013;79:347–360.
- Watabe-Uchida M., Zhu L., Ogawa S.K., Vamanrao A., Uchida N. Whole-brain mapping of direct inputs to midbrain dopamine neurons. Neuron. 2012;74:858–873.
- Wickersham I.R., Lyon D.C., Barnard R.J., Mori T., Finke S., Conzelmann K.K., Young J.A., Callaway E.M. Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons. Neuron. 2007;53:639–647.
- Zufferey R., Nagy D., Mandel R.J., Naldini L., Trono D. Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nat. Biotechnol. 1997;15:871–875.
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