Anchored FRET sensors detect local caspase activation prior to neuronal degeneration

Molecular Neurodegeneration

Einar Hallberg¹  Kerstin Iverfeldt¹  Mu Zhang¹  Malin Samuelsson¹  Tom Gatsinzi¹  Veronica Ramberg¹  Ricardo A Figueroa² 

[1] Department of Neurochemistry, Stockholm University, SE-106 91 Stockholm, Sweden;Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden

关键词: Spatiotemporal analysis;    Neurodegeneration;    Neurite degeneration;    Live Cell Imaging;    FRET;    Caspases;    Amyloid-β;   

Others  :  865845
DOI  :  10.1186/1750-1326-6-35

 received in 2010-10-05, accepted in 2011-05-23,  发布年份 2011

【 摘 要 】

Background

Recent studies indicate local caspase activation in dendrites or axons during development and in neurodegenerative disorders such as Alzheimer's disease (AD). Emerging evidences point to soluble oligomeric amyloid-β peptide as a causative agent in AD.

Results

Here we describe the design of fluorescence resonance energy transfer (FRET)-based caspase sensors, fused to the microtubule associated protein tau. Specific caspase sensors preferentially cleaved by caspase-3, -6 or -9 were expressed in differentiated human neuroblastoma SH-SY5Y cells. The anchoring of the sensors resulted in high FRET signals both in extended neurites and soma and made analysis of spatiotemporal signal propagation possible. Caspase activation was detected as loss of FRET after exposure to different stimuli. Interestingly, after staurosporine treatment caspase-6 activation was significantly delayed in neurites compared to cell bodies. In addition, we show that exposure to oligomer-enriched amyloid-β peptide resulted in loss of FRET in cells expressing sensors for caspase-3 and -6, but not -9, in both soma and neurites before neurite degeneration was observed.

Conclusions

Taken together, the results show that by using anchored FRET sensors it is possible to detect stimuli-dependent differential activation of caspases and to distinguish local from global caspase activation in live neuronal cells. Furthermore, in these cells oligomer-enriched amyloid-β peptide induces a global, rather than local activation of caspase-3 and -6, which subsequently leads to neuronal cell death.

【 授权许可】

   
2011 Figueroa et al; licensee BioMed Central Ltd.

【 图 表 】

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