Abstract
There is now crucial medical importance placed on understanding the role of early stage, subvisible protein aggregation, particularly in neurodegenerative disease. While there are strategies for detecting such aggregates in vitro, there is no approach at present that can detect these toxic species associated with cells and specific subcellular compartments. We have exploited excitation-energy-dependent fluorescence edge-shift of recombinant protein labeled with a molecular beacon, to provide a sensitive read out for the presence of subvisible protein aggregates. To demonstrate the potential utility of the approach, we examine the major peptide associated with the initiation of Alzheimer's disease, amyloid β-protein (Aβ) at a patho-physiologically relevant concentration in mouse cortical neurons. Using our approach, we find preliminary evidence that subvisible Aβ aggregates are detected at specific subcellular regions and that neurons drive the formation of specific Aβ aggregate conformations. These findings therefore demonstrate the potential of a novel fluorescence-based approach for detecting and imaging protein aggregates in a cellular context, which can be used to sensitively probe the association of early stage toxic protein aggregates within subcellular compartments.
| Original language | English |
|---|---|
| Pages (from-to) | 1240-1250 |
| Number of pages | 11 |
| Journal | ACS Chemical Neuroscience |
| Volume | 10 |
| Issue number | 3 |
| Early online date | 16 Oct 2018 |
| DOIs | |
| Publication status | Published - 20 Mar 2019 |
Keywords
- Alzheimer's
- amyloid beta
- edge-shift
- Fluorescence
- neuron
ASJC Scopus subject areas
- Biochemistry
- Physiology
- Cognitive Neuroscience
- Cell Biology
Cite this
Excitation-energy-dependent molecular beacon detects early stage neurotoxic aβ aggregates in the presence of cortical neurons. / Gulácsy, Christina E.; Meade, Richard; Catici, Dragana A.M.; Soeller, Christian; Pantos, G. Dan; Jones, D. Dafydd; Alibhai, Dominic; Jepson, Mark; Valev, Ventsislav K.; Mason, Jody M.; Williams, Robert J.; Pudney, Christopher R.
In: ACS Chemical Neuroscience, Vol. 10, No. 3, 20.03.2019, p. 1240-1250.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Excitation-energy-dependent molecular beacon detects early stage neurotoxic aβ aggregates in the presence of cortical neurons
AU - Gulácsy, Christina E.
AU - Meade, Richard
AU - Catici, Dragana A.M.
AU - Soeller, Christian
AU - Pantos, G. Dan
AU - Jones, D. Dafydd
AU - Alibhai, Dominic
AU - Jepson, Mark
AU - Valev, Ventsislav K.
AU - Mason, Jody M.
AU - Williams, Robert J.
AU - Pudney, Christopher R.
PY - 2019/3/20
Y1 - 2019/3/20
N2 - There is now crucial medical importance placed on understanding the role of early stage, subvisible protein aggregation, particularly in neurodegenerative disease. While there are strategies for detecting such aggregates in vitro, there is no approach at present that can detect these toxic species associated with cells and specific subcellular compartments. We have exploited excitation-energy-dependent fluorescence edge-shift of recombinant protein labeled with a molecular beacon, to provide a sensitive read out for the presence of subvisible protein aggregates. To demonstrate the potential utility of the approach, we examine the major peptide associated with the initiation of Alzheimer's disease, amyloid β-protein (Aβ) at a patho-physiologically relevant concentration in mouse cortical neurons. Using our approach, we find preliminary evidence that subvisible Aβ aggregates are detected at specific subcellular regions and that neurons drive the formation of specific Aβ aggregate conformations. These findings therefore demonstrate the potential of a novel fluorescence-based approach for detecting and imaging protein aggregates in a cellular context, which can be used to sensitively probe the association of early stage toxic protein aggregates within subcellular compartments.
AB - There is now crucial medical importance placed on understanding the role of early stage, subvisible protein aggregation, particularly in neurodegenerative disease. While there are strategies for detecting such aggregates in vitro, there is no approach at present that can detect these toxic species associated with cells and specific subcellular compartments. We have exploited excitation-energy-dependent fluorescence edge-shift of recombinant protein labeled with a molecular beacon, to provide a sensitive read out for the presence of subvisible protein aggregates. To demonstrate the potential utility of the approach, we examine the major peptide associated with the initiation of Alzheimer's disease, amyloid β-protein (Aβ) at a patho-physiologically relevant concentration in mouse cortical neurons. Using our approach, we find preliminary evidence that subvisible Aβ aggregates are detected at specific subcellular regions and that neurons drive the formation of specific Aβ aggregate conformations. These findings therefore demonstrate the potential of a novel fluorescence-based approach for detecting and imaging protein aggregates in a cellular context, which can be used to sensitively probe the association of early stage toxic protein aggregates within subcellular compartments.
KW - Alzheimer's
KW - amyloid beta
KW - edge-shift
KW - Fluorescence
KW - neuron
UR - http://www.scopus.com/inward/record.url?scp=85057871010&partnerID=8YFLogxK
U2 - 10.1021/acschemneuro.8b00322
DO - 10.1021/acschemneuro.8b00322
M3 - Article
VL - 10
SP - 1240
EP - 1250
JO - ACS Chemical Neuroscience
JF - ACS Chemical Neuroscience
SN - 1948-7193
IS - 3
ER -