Imaging Nanometer-Sized α-Synuclein Aggregates by Superresolution Fluorescence Localization Microscopy

Roberti, M. Julia; Fölling, Jonas; Celej, M. Soledad; Bossi, Mariano L.; Jovin, Thomas M.; Jares‐Erijman, Elizabeth A.

doi:10.1016/j.bpj.2012.03.010

Cited by 60 publications

(52 citation statements)

References 57 publications

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“…For example, seeded aggregates of Aβ taken up by HeLa cells were imaged by the dSTORM method11, but it remained unclear if the aggregation occurred in the living cells or whether larger aggregates were, at least partially, endocytosed into the cell interior. The in vitro and in-cell Aβ aggregates had similar morphology, in contrast to results for α-synuclein, where aggregates were observed after microinjection of protein into cells12.…”

mentioning

confidence: 51%

Cellular Inclusion Bodies of Mutant Huntingtin Exon 1 Obscure Small Fibrillar Aggregate Species

Sahl

Weiss

Duim

et al. 2012

Sci Rep

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The identities of toxic aggregate species in Huntington's disease pathogenesis remain ambiguous. While polyQ-expanded huntingtin (Htt) is known to accumulate in compact inclusion bodies inside neurons, this is widely thought to be a protective coping response that sequesters misfolded conformations or aggregated states of the mutated protein. To define the spatial distributions of fluorescently-labeled Htt-exon1 species in the cell model PC12m, we employed highly sensitive single-molecule super-resolution fluorescence imaging. In addition to inclusion bodies and the diffuse pool of monomers and oligomers, fibrillar aggregates ~100 nm in diameter and up to ~1–2 µm in length were observed for pathogenic polyQ tracts (46 and 97 repeats) after targeted photo-bleaching of the inclusion bodies. These short structures bear a striking resemblance to fibers described in vitro. Definition of the diverse Htt structures in cells will provide an avenue to link the impact of therapeutic agents to aggregate populations and morphologies.

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confidence: 51%

Cellular Inclusion Bodies of Mutant Huntingtin Exon 1 Obscure Small Fibrillar Aggregate Species

Sahl

Weiss

Duim

et al. 2012

Sci Rep

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“…54 In general, spherical colloidal assemblies are forced into contact when the respective streamlines of the solvent get closer than the diameter of the particles, such behavior has been shown to be the dominant cause for aggregation of particles in the micrometer range. 60,61 Considering that gut environment is highly dynamic, 62 the feasibility of the 33-mer to form colloidal nanospheres which can further organize into linear aggregates depending on environmental conditions would be relevant; however, in vivo studies are required to validate the real relevance. Once the colloidal nanoparticles are formed, their intrinsic characteristics and the medium condition contribute to random assembly, as it would be expected for globular particles or spontaneous linear organization and formation of fibrils (Figure 9).…”

Section: Supramolecular Organization Of 33-mer Detected By Transmissimentioning

confidence: 99%

Circular dichroism and electron microscopy studies in vitro of 33‐mer gliadin peptide revealed secondary structure transition and supramolecular organization

Herrera

Zamarreño²,

Costabel³

et al. 2013

Biopolymers

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Gliadin, a protein present in wheat, rye, and barley, undergoes incomplete enzymatic degradation during digestion, producing an immunogenic 33-mer peptide, LQLQPF(PQPQLPY)3 PQPQPF. The special features of 33-mer that provoke a break in its tolerance leading to gliadin sensitivity and celiac disease remains elusive. Herein, it is reported that 33-mer gliadin peptide was not only able to fold into polyproline II secondary structure but also depending on concentration resulted in conformational transition and self-assembly under aqueous condition, pH 7.0. A 33-mer dimer is presented as one initial possible step in the self-assembling process obtained by partial electrostatics charge distribution calculation and molecular dynamics. In addition, electron microscopy experiments revealed supramolecular organization of 33-mer into colloidal nanospheres. In the presence of 1 mM sodium citrate, 1 mM sodium borate, 1 mM sodium phosphate buffer, 15 mM NaCl, the nanospheres were stabilized, whereas in water, a linear organization and formation of fibrils were observed. It is hypothesized that the self-assembling process could be the result of the combination of hydrophobic effect, intramolecular hydrogen bonding, and electrostatic complementarity due to 33-mer's high content of proline and glutamine amino acids and its calculated nonionic amphiphilic character. Although, performed in vitro, these experiments have revealed new features of the 33-mer gliadin peptide that could represent an important and unprecedented event in the early stage of 33-mer interaction with the gut mucosa prior to onset of inflammation. Moreover, these findings may open new perspectives for the understanding and treatment of gliadin intolerance disorders.

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“…We used induced pluripotent stem cells (iPSCs) from a PD patient with a triplication of the SNCA gene and from a healthy control unaffected by the disease to generate cortical neurons. Although SR methods have been used to image fibrils in cells, these are typically exogenously added aggregates generated from fluorophore‐labelled protein 28, 43, 44, 45, 46, 47. This is the first case in which a specific probe for aggregates has been used, and it enables the SR imaging of unlabelled, endogenous aberrant protein complexes.…”

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confidence: 99%

Nanoscopic Characterisation of Individual Endogenous Protein Aggregates in Human Neuronal Cells

et al. 2018

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The aberrant misfolding and subsequent conversion of monomeric protein into amyloid aggregates characterises many neurodegenerative disorders, including Parkinson's and Alzheimer's diseases. These aggregates are highly heterogeneous in structure, generally of low abundance and typically smaller than the diffraction limit of light (≈250 nm). To overcome the challenges these characteristics pose to the study of endogenous aggregates formed in cells, we have developed a method to characterise them at the nanometre scale without the need for a conjugated fluorophore. Using a combination of DNA PAINT and an amyloid‐specific aptamer, we demonstrate that this technique is able to detect and super‐resolve a range of aggregated species, including those formed by α‐synuclein and amyloid‐β. Additionally, this method enables endogenous protein aggregates within cells to be characterised. We found that neuronal cells derived from patients with Parkinson's disease contain a larger number of protein aggregates than those from healthy controls.

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Imaging Nanometer-Sized α-Synuclein Aggregates by Superresolution Fluorescence Localization Microscopy

Cited by 60 publications

References 57 publications

Cellular Inclusion Bodies of Mutant Huntingtin Exon 1 Obscure Small Fibrillar Aggregate Species

Cellular Inclusion Bodies of Mutant Huntingtin Exon 1 Obscure Small Fibrillar Aggregate Species

Circular dichroism and electron microscopy studies in vitro of 33‐mer gliadin peptide revealed secondary structure transition and supramolecular organization

Nanoscopic Characterisation of Individual Endogenous Protein Aggregates in Human Neuronal Cells

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