In vivo imaging of α-synuclein in mouse cortex demonstrates stable expression and differential subcellular compartment mobility

Vivek K. Unni, Tamily A. Weissman, Edward Rockenstein, Eliezer Masliah, Pamela J McLean, Bradley T. Hyman

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background: Regulation of α-synuclein levels within cells is thought to play a critical role in Parkinson's Disease (PD) pathogenesis and in other related synucleinopathies. These processes have been studied primarily in reduced preparations, including cell culture. We now develop methods to measure α-synuclein levels in the living mammalian brain to study in vivo protein mobility, turnover and degradation with subcellular specificity. Methodology/Principal Findings: We have developed a system using enhanced Green Fluorescent Protein (GFP)-tagged human α-synuclein (Syn-GFP) transgenic mice and in vivo multiphoton imaging to measure α-synuclein levels with subcellular resolution. This new experimental paradigm allows individual Syn-GFP-expressing neurons and presynaptic terminals to be imaged in the living mouse brain over a period of months. We find that Syn-GFP is stably expressed by neurons and presynaptic terminals over this time frame and further find that different presynaptic terminals can express widely differing levels of Syn-GFP. Using the fluorescence recovery after photobleaching (FRAP) technique in vivo we provide evidence that at least two pools of Syn-GFP exist in terminals with lower levels of mobility than measured previously. These results demonstrate that multiphoton imaging in Syn-GFP mice is an excellent new strategy for exploring the biology of α-synuclein and related mechanisms of neurodegeneration. Conclusions/Significance: In vivo multiphoton imaging in Syn-GFP transgenic mice demonstrates stable α-synuclein expression and differential subcellular compartment mobility within cortical neurons. This opens new avenues for studying α-synuclein biology in the living brain and testing new therapeutics for PD and related disorders.

Original languageEnglish (US)
Article numbere10589
JournalPLoS One
Volume5
Issue number5
DOIs
StatePublished - 2010
Externally publishedYes

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Synucleins
Green Fluorescent Proteins
green fluorescent protein
cortex
image analysis
Imaging techniques
mice
Presynaptic Terminals
Neurons
Brain
Parkinson disease
neurons
Transgenic Mice
Parkinson Disease
brain
Fluorescence Recovery After Photobleaching
Photobleaching
genetically modified organisms
Biological Sciences
Cell culture

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

In vivo imaging of α-synuclein in mouse cortex demonstrates stable expression and differential subcellular compartment mobility. / Unni, Vivek K.; Weissman, Tamily A.; Rockenstein, Edward; Masliah, Eliezer; McLean, Pamela J; Hyman, Bradley T.

In: PLoS One, Vol. 5, No. 5, e10589, 2010.

Research output: Contribution to journalArticle

Unni, Vivek K. ; Weissman, Tamily A. ; Rockenstein, Edward ; Masliah, Eliezer ; McLean, Pamela J ; Hyman, Bradley T. / In vivo imaging of α-synuclein in mouse cortex demonstrates stable expression and differential subcellular compartment mobility. In: PLoS One. 2010 ; Vol. 5, No. 5.
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