Lentivirus-based genetic manipulations of cortical neurons and their optical and electrophysiological monitoring in vivo

Dittgen, T., Nimmerjahn, A., Komai, S., Licznerski, P., Waters, J., Margrie, T. W., Helmchen, F., Denk, W., Brecht, M., Osten, P. (December 2004) Lentivirus-based genetic manipulations of cortical neurons and their optical and electrophysiological monitoring in vivo. Proceedings of the National Academy of Sciences of the United States of America, 101 (52). pp. 18206-11. ISSN 0027-8424

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URL: http://www.ncbi.nlm.nih.gov/pubmed/15608064
DOI: 10.1073/pnas.0407976101

Abstract

It is becoming increasingly clear that single cortical neurons encode complex and behaviorally relevant signals, but efficient means to study gene functions in small networks and single neurons in vivo are still lacking. Here, we establish a method for genetic manipulation and subsequent phenotypic analysis of individual cortical neurons in vivo. First, lentiviral vectors are used for neuron-specific gene delivery from alpha-calcium/calmodulin-dependent protein kinase II or Synapsin I promoters, optionally in combination with gene knockdown by means of U6 promoter-driven expression of short-interfering RNAs. Second, the phenotypic analysis at the level of single cortical cells is carried out by using two-photon microscopy-based techniques: high-resolution two-photon time-lapse imaging is used to monitor structural dynamics of dendritic spines and axonal projections, whereas cellular response properties are analyzed electrophysiologically by two-photon microscopy directed whole-cell recordings. This approach is ideally suited for analysis of gene functions in individual neurons in the intact brain.

Item Type: Paper
Uncontrolled Keywords: Animals Calcium Calmodulin Dependent Protein Kinase Type 2 Calcium Calmodulin Dependent Protein Kinases genetics Electrophysiology methods Genetic Vectors Green Fluorescent Proteins metabolism Hippocampus metabolism Image Processing Computer Assisted Lentivirus genetics Mice Mice, Inbred C57BL Microscopy Neurons metabolism Phenotype Photons Promoter Regions Genetic RNA Small Interfering metabolism RNA Small Nuclear metabolism Recombinant Proteins genetics metabolism Synapsins genetics
Subjects: organism description > virus > lentivirus
therapies > viral vectors
CSHL Authors:
Communities: CSHL labs > Osten lab
Depositing User: CSHL Librarian
Date: 28 December 2004
Date Deposited: 09 Feb 2012 17:55
Last Modified: 11 Jan 2018 20:45
PMCID: PMC539748
Related URLs:
URI: http://repository.cshl.edu/id/eprint/22364

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