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

Dittgen, T., Nimmerjahn, A., Komai, S., Licznerski, P., Waters, D. J., Margrie, T. W., et al. (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), 18206-18211. doi:10.1073/pnas.0407976101.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-15C7-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-15C8-5
Genre: Journal Article
Alternative Title : Lentivirus-based genetic manipulations of cortical neurons and their optical and electrophysiological monitoring in vivo

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PNAS_101_2004_18206.pdf (Any fulltext), 401KB
 
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 Creators:
Dittgen, Tanjew1, Author              
Nimmerjahn, Axel2, Author              
Komai, Shoji3, Author              
Licznerski, Pawel1, Author              
Waters, David Jack2, Author              
Margrie, Troy W.1, 2, Author              
Helmchen, Fritjof2, Author              
Denk, Winfried3, Author              
Brecht, Michael, Author
Osten, Pavel1, Author              
Affiliations:
1Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497704              
2Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497701              
3Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497699              

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Free keywords: patch-clamp recording two-photon imaging
 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.

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Language(s): eng - English
 Dates: 2004-08-262004-11-042004-12-172004-12-28
 Publication Status: Published in print
 Pages: 6
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
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Title: Proceedings of the National Academy of Sciences of the United States of America
  Other : Proc. Natl. Acad. Sci. U. S. A.
Source Genre: Journal
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Publ. Info: National Academy of Sciences
Pages: - Volume / Issue: 101 (52) Sequence Number: - Start / End Page: 18206 - 18211 Identifier: ISSN: 0027-8424
CoNE: https://pure.mpg.de/cone/journals/resource/954925427230