title: Simultaneous visualization of multiple neuronal properties with single-cell resolution in the living rodent brain.
abstract: To understand the fine-scale structures and functional properties of individual neurons in vivo, we developed and validated a rapid genetic technique that enables simultaneous investigation of multiple neuronal properties with single-cell resolution in the living rodent brain. Our technique PASME (promoter-assisted sparse-neuron multiple-gene labeling using in uteroelectroporation) targets specific small subsets of sparse pyramidal neurons in layer 2/3, layer 5 of the cerebral cortex and in the hippocampus with multiple fluorescent reporter proteins such as postsynaptic PSD-95-GFP and GFP-gephyrin. The technique is also applicable for targeting independently individual neurons and their presynaptic inputs derived from surrounding neurons. Targeting sparse layer 2/3 neurons, we uncovered a novel subpopulation of layer 2/3 neurons in the mouse cerebral cortex. This technique, broadly applicable for probing and manipulating neurons with single-cell resolution in vivo, should provide a robust means to uncover the basic mechanisms employed by the brain, especially when combined with in vivo two-photon laser-scanning microscopy and/or optogenetic technologies.
authors: Department of Molecular and Systems Neurobiology, Graduate School of Medicine, The University of Tokyo, Japan.
authors: Rie
authors: R
authors: Ako
authors: Mayu
authors: M
authors: Wakimoto
authors: Haruka
authors: H
authors: Ebisu
authors: Kaori
authors: K
authors: Tanno
authors: Riichiro
authors: R
authors: Hira
authors: Haruo
authors: H
authors: Kasai
authors: Masanori
authors: M
authors: Matsuzaki
authors: Hiroshi
authors: H
authors: Kawasaki
mesh_terms: D000818:Animals; D002452:Cell Count; D002540:Cerebral Cortex / Q000166:cytology / Q000502:physiology*; D018274:Electroporation / Q000379:methods; D018014:Gene Transfer Techniques; D051379:Mice; D008813:Mice, Inbred ICR; D018613:Microscopy, Confocal / Q000379:methods; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar