title: Targeting single neuronal networks for gene expression and cell labeling in vivo.
abstract: To understand fine-scale structure and function of single mammalian neuronal networks, we developed and validated a strategy to genetically target and trace monosynaptic inputs to a single neuron in vitro and in vivo. The strategy independently targets a neuron and its presynaptic network for specific gene expression and fine-scale labeling, using single-cell electroporation of DNA to target infection and monosynaptic retrograde spread of a genetically modifiable rabies virus. The technique is highly reliable, with transsynaptic labeling occurring in every electroporated neuron infected by the virus. Targeting single neocortical neuronal networks in vivo, we found clusters of both spiny and aspiny neurons surrounding the electroporated neuron in each case, in addition to intricately labeled distal cortical and subcortical inputs. This technique, broadly applicable for probing and manipulating single neuronal networks with single-cell resolution in vivo, may help shed new light on fundamental mechanisms underlying circuit development and information processing by neuronal networks throughout the brain.
authors.affiliation: Systems Neurobiology Laboratories, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: James H
authors.initials: JH
authors.lastname: Marshel
authors.forename: Takuma
authors.initials: T
authors.lastname: Mori
authors.forename: Kristina J
authors.initials: KJ
authors.lastname: Nielsen
authors.forename: Edward M
authors.initials: EM
authors.lastname: Callaway
mesh_terms: D000818:Animals; D018274:Electroporation; D015870:Gene Expression; D005822:Genetic Vectors; D006652:Histological Techniques; D066298:In Vitro Techniques; D051379:Mice; D019579:Neocortex / Q000166:cytology* / Q000378:metabolism / Q000821:virology; D009434:Neural Pathways / Q000166:cytology / Q000502:physiology*; D056347:Neuroanatomical Tract-Tracing Techniques / Q000379:methods*; D056325:Neuronal Tract-Tracers; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000502:physiology* / Q000821:virology; D017729:Presynaptic Terminals / Q000378:metabolism / Q000821:virology; D017966:Pyramidal Cells / Q000166:cytology / Q000502:physiology; D011820:Rabies virus / Q000235:genetics; D051381:Rats; D015203:Reproducibility of Results; D014793:Visual Cortex / Q000166:cytology / Q000378:metabolism / Q000821:virology
chemical_list: D056325:Neuronal Tract-Tracers
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