Chyi Wei Chung's research while affiliated with University of Cambridge and other places
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Publications (15)
During aging, proteostasis capacity declines and distinct proteins become unstable and can accumulate as protein aggregates inside and outside of cells. Both in disease and during aging, proteins selectively aggregate in certain tissues and not others. Yet, tissue-specific regulation of cytoplasmic protein aggregation remains poorly understood. Sur...
Monomeric alpha-synuclein (aSyn) is a well characterised protein that importantly binds to lipids. aSyn monomers assemble into amyloid fibrils which are localised to lipids and organelles in insoluble structures found in Parkinson’s disease patient’s brains. Previous work to address pathological aSyn-lipid interactions has focused on using syntheti...
Structured Illumination Microscopy, SIM, is one of the most powerful optical imaging methods available to visualize biological environments at subcellular resolution. Its limitations stem from a difficulty of imaging in multiple color channels at once, which reduces imaging speed. Furthermore, there is substantial experimental complexity in setting...
Neurodegenerative diseases are associated with protein misfolding and amyloid aggregation. The work presented in this thesis involves the use of fluorescence lifetime imaging microscopy (FLIM) and other biophysical techniques to elucidate the molecular mechanisms that underlie different amyloid-associated neurodegenerative dis- eases. Currently, th...
The solvation shell is essential for the folding and function of proteins, but how it contributes to protein misfolding and aggregation has still to be elucidated. We show that the mobility of solvation shell H2O molecules influences the aggregation rate of the amyloid protein α‐synuclein (αSyn), a protein associated with Parkinson’s disease. When...
Monomeric alpha-synuclein (aSyn) is a well characterised as a lipid binding protein. aSyn is known to form amyloid fibrils which are also localised with lipids and organelles in so called Lewy bodies, insoluble structures found in Parkinson s disease patient s brains. It is still unclear under which conditions the aSyn-lipid interaction can start t...
The molecular mechanisms that connect the formation of aberrant cytoplasmic FUS condensates to biological malfunction are incompletely understood. Here, we develop an approach to determine the intracellular FUS viscosity in live mammalian cells and find that ALS-related mutant P525L-FUS forms the most viscous condensates and has impaired cytoskelet...
Neurodegenerative disorders are a family of diseases that remain poorly treated despite their growing global health burden. A shared feature of many neurodegenerative disorders is the accumulation of toxic misfolded proteins. To gain insight into the mechanisms and modulators of protein misfolding, we developed a multiplex reverse genetics platform...
Sub-diffraction resolution, gentle sample illumination, and the possibility to image in multiple colors make Structured Illumination Microscopy (SIM) an imaging technique which is particularly well suited for live cell observations. Here, we present Machine learning Assisted Interferometric-SIM (MAI-SIM), an easy-to-implement method for high speed...
The aggregation of Aβ42 is a hallmark of Alzheimer's disease. It is still not known what the biochemical changes are inside a cell which will eventually lead to Aβ42 aggregation. Thermogenesis has been associated with cellular stress, the latter of which may promote aggregation. We perform intracellular thermometry measurements using fluorescent po...
The aggregation of Aβ42 is a hallmark of Alzheimer′s disease. It is still not known what the biochemical changes are inside a cell which will eventually lead to Aβ42 aggregation. Thermogenesis has been associated with cellular stress, the latter of which may promote aggregation. We perform intracellular thermometry measurements using fluorescent po...
Conventional in vitro aggregation assays often involve tagging with extrinsic fluorophores, which can interfere with aggregation. We propose the use of intrinsic amyloid fluorescence lifetime probed using two-photon excitation and represented by model-free phasor plots as a label-free assay to characterize the amyloid structure. Intrinsic amyloid f...
Conventional in vitro aggregation assays often involve tagging with extrinsic fluorophores which can interfere with aggregation. We propose the use of intrinsic amyloid fluorescence lifetime represented by model-free phasor plots, as a label-free assay to characterise amyloid structure. Intrinsic amyloid fluorescence arises from structured packing...
Temperature is a fundamental physical parameter that influences biological processes in living cells. Hence, intracellular temperature mapping can be used to derive useful information reflective of thermodynamic properties and cellular behaviour. Here, existing publications of different thermometry systems, focusing on those that employ fluorescenc...
Citations
... Specific autophagic genes and their roles in aging and disease phenotypes have recently been reviewed in more detail [25]. Besides aging, increasing autophagy and the UPS have been shown to improve health in animal models with protein aggregate-type diseases [7,[26][27][28][29][30]. ...
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... These studies have revealed that exogenous α-synuclein induces Lewy body pathology and lead to a decrease in synaptic protein, reduced synaptic activity, and altered spine dynamics prior to neuron loss [11,12,24]. Some studies have shown that α-synuclein PFFs and oligomers directly interact with membranes and synaptic vesicles, modulating calcium transients and vesicle homeostasis [25][26][27]. Here, we assessed the synaptic changes that occur during the propagation of α-synuclein in a neural network by measuring sEPSCs in the somatosensory cortex after PFF injection into the striatum, finding a decrease in the frequency of spontaneous events and a concomitant decrease in synapse density. ...
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... However, in practice, the equivalent modulation depth of the raw data is largely determined by the modulation depth of the illumination patterns, as well as the modulation contrast and SNR of the samples. So, well-aligned light paths, and illumination patterns with precise phase shift and minimal distortion [47,48] are still crucial for high-quality reconstruction of direct-SIM. Raw data with high SNR and high modulation contrast is beneficial for high-quality reconstruction of direct-SIM. ...
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... Hence, changing the isotopic composition of the water can be used to modulate collagen-water interactions, and so study their effect on the assembly process without affecting the electrostatic interactions due to changes in the solvent dielectric constants. A significant effect of D 2 O on protein self assembly has been recently observed for αsynuclein (aS) and insulin (INS) (16,17). In these studies, it was suggested that in D 2 O specific folded structures are stabilized, accelerating (in the case of aS) or slowing down (in the case of INS) the assembly. ...
... It is orthologous to human DnaJC11 (DnaJ heat shock protein family (Hsp40) member C11). DnaJC11 is involved in cristae formation in the mitochondrion [39][40][41]. CG43322 is predicted to be involved in the Golgi organization, it is orthologous to human DnaJC28 (DnaJ heat shock protein family (Hsp40) member C28) and is present in the Golgi transport complex. ...
... There are reports on cellular and intracellular temperature (T) gradients, produced and potentially sustained via cell metabolic activity, in different cell models, tissues or cancer models [157][158][159][160]. The field of nanothermometry addresses such measurements via FLIM, PLIM and dedicated small molecule and nanosensor probes [161][162][163]. ...
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... 19−21 There are some examples of the advantages of FLIM in the study of biological systems, 22 namely, in the study of aggregation/disaggregation of amyloid proteins. 23 Nevertheless, the use of this technique is still rare in the field of materials science. 24 In this work, we aim to use FLIM to examine solvent-driven self-assembled structures of porphyrin bearing triphenylamine substituents, meso-tetra-p-(di-p-phenylamino)phenylporphyrin, H 2 T(TPA) 4 P (inset of Figure 1b and Scheme 1), and the characterization of the film morphology. ...
... Although this renders some polymorphs invisible in FLIM and precludes their analysis, it indirectly reflects differences in the amino acid residues that constitute the core of the fibers or the stacking of the individual monomers, and thus enables the discrimination of conformational variants. It would be interesting to compare the different α-Syn polymorphs in two-photon FLIM, which does not require labeling to examine whether this approach also allows their differentiation (Chung et al., 2021). ...
... Temperature measurements with nanoscale resolution can provide unique information such as metabolic functions and life cycles within cells. 1,2 The development of nanothermometers is therefore critical for thermal biology. Diamond nanoparticles, also known as nanodiamonds (NDs), containing various fluorescent defects, have been proposed as promising nanothermometers due to their excellent thermal conductivity, remarkable photostability, attractive biocompatibility, and extreme chemical inertness. ...
