Viral Applications of Green Fluorescent Protein pp 63-95

Part of the Methods in Molecular Biology™ book series (MIMB, volume 515)

Use of GFP to Analyze Morphology, Connectivity, and Function of Cells in the Central Nervous System

  • Alan R. Harvey
  • Erich Ehlert
  • Joris de Wit
  • Eleanor S. Drummond
  • Margaret A. Pollett
  • Marc Ruitenberg
  • Giles W. Plant
  • Joost Verhaagen
  • Christiaan N. Levelt

Summary

We here describe various approaches using GFP that are being used in the morphological and functional analysis of specific cell types in the normal and injured central nervous system. Incorporation of GFP into viral vectors allows phenotypic characterization of transduced cells and can be used to label their axons and terminal projections. Characterization of transduced cell morphology can be enhanced by intracellular injection of living GFP-labeled cells with appropriate fluorescent dyes. Ex vivo labeling of precursor or glial cells using viral vectors that encode GFP permits long-term identification of these cells after transplantation into the brain or spinal cord. In utero electroporation methods result in expression of gene products in developing animals, allowing both functional and morphological studies to be carried out. GFPCre has been developed as a marker gene for viral vector-mediated expression of the bacterial recombinase Cre in the brain of adult mice with “floxed” transgenes. GFPCre-mediated induction of transgene expression can be monitored by GFP expression in defined populations of neurons in the adult brain. Finally, GFP can be used to tag proteins, permitting dynamic visualization of the protein of interest in living cells.

Key words

Viral vectors Adeno-associated virus Lentivirus Electroporation Olfactory ensheathing glia Schwann cells Cre-EGFP Retina Semaphorin Hippocampus Neocortex Spinal cord 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Alan R. Harvey
    • 1
  • Erich Ehlert
    • 2
  • Joris de Wit
    • 3
  • Eleanor S. Drummond
    • 1
  • Margaret A. Pollett
    • 1
  • Marc Ruitenberg
    • 4
  • Giles W. Plant
    • 4
  • Joost Verhaagen
    • 2
  • Christiaan N. Levelt
    • 5
  1. 1.School of Anatomy and Human BiologyThe University of Western AustraliaCrawleyAustralia
  2. 2.Netherlands Institute for NeuroscienceRoyal Netherlands Academy of Arts and SciencesAmsterdamthe Netherlands
  3. 3.Division of Biology, Neurobiology SectionUniversity of California San DiegoLa JollaUSA
  4. 4.School of Anatomy and Human Biology and Red’s Spinal Cord Research LaboratoryThe University of Western AustraliaCrawleyAustralia
  5. 5.Research Group Molecular Visual PlasticityNetherlands Institute for Neuroscience, Royal Netherlands Academy of Arts and SciencesAmsterdamthe Netherlands

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