A5041913324- Advanced Fluorescence Microscopy Techniques
- Parkinson's Disease Mechanisms and Treatments
- Alzheimer's disease research and treatments
- Advanced Electron Microscopy Techniques and Applications
- Cell Image Analysis Techniques
- Neurological disorders and treatments
- Advanced Biosensing Techniques and Applications
- Mitochondrial Function and Pathology
- Neuroscience and Neuropharmacology Research
- Amyotrophic Lateral Sclerosis Research
- Genetic Neurodegenerative Diseases
- Photoreceptor and optogenetics research
- Advanced biosensing and bioanalysis techniques
- Botulinum Toxin and Related Neurological Disorders
- Neurogenetic and Muscular Disorders Research
- Nuclear Receptors and Signaling
- RNA Interference and Gene Delivery
- RNA Research and Splicing
- Prion Diseases and Protein Misfolding
- Nuclear Engineering Thermal-Hydraulics
- Pluripotent Stem Cells Research
- RNA modifications and cancer
- Monoclonal and Polyclonal Antibodies Research
- Cellular transport and secretion
- Nanoplatforms for cancer theranostics
University of Edinburgh
2018-2025
MRC Centre for Regenerative Medicine
2023-2025
European Molecular Biology Laboratory
2024
Centre for Inflammation Research
2024
Institute of Genetics and Cancer
2024
Bridge University
2023
University of Cambridge
2011-2020
UK Dementia Research Institute
2018-2019
National Hospital for Neurology and Neurosurgery
2018
University College London
2018
Protein aggregation causes α-synuclein to switch from its physiological role a pathological toxic gain of function. Under conditions, monomeric improves ATP synthase efficiency. Here, we report that monomers generates beta sheet-rich oligomers localise the mitochondria in close proximity several mitochondrial proteins including synthase. Oligomeric impairs complex I-dependent respiration. Oligomers induce selective oxidation subunit and lipid peroxidation. These events increase probability...
Protein aggregation and oxidative stress are both key pathogenic processes in Parkinson's disease, although the mechanism by which misfolded proteins induce neuronal death remains unknown. In this study, we describe how of alpha-synuclein (α-S) from its monomeric form to soluble oligomeric state results aberrant free radical production toxicity.We first demonstrate excessive a human induced pluripotent stem-derived α-S triplication model at basal levels on application picomolar doses...
Significance Growing experimental evidence suggests that the pathological spreading of alpha-synuclein aggregates in Parkinson’s disease is mediated through a process templated seeding whereby catalyze conversion soluble protein molecules into their aggregated forms. A molecular-level understanding this still lacking. Here, we determine concentrations and numbers necessary for effective alpha-synuclein, thus providing quantitative framework to understand conditions when its seeded...
Abstract Protein aggregation and abnormal lipid homeostasis are both implicated in neurodegeneration through unknown mechanisms. Here we demonstrate that aggregate-membrane interaction is critical to induce a form of cell death called ferroptosis. Importantly, the drives ferroptosis depends on conformational structure aggregate, as well oxidation state membrane. We generated human stem cell-derived models synucleinopathy, characterized by intracellular formation α-synuclein aggregates bind...
Abstract Protein aggregation plays a key role in neurodegenerative disease, giving rise to small oligomers that may become cytotoxic cells. The fundamental microscopic reactions taking place during aggregation, and their rate constants, have been difficult determine due lack of suitable methods identify follow the low concentration over time. Here we use single-molecule fluorescence study repeat domain tau (K18), two mutant forms linked with familial frontotemporal dementia, deletion ΔK280...
Abstract Super-resolution microscopy allows biological systems to be studied at the nanoscale, but has been restricted providing only positional information. Here, we show that it is possible perform multi-dimensional super-resolution imaging determine both position and environmental properties of single-molecule fluorescent emitters. The method presented here exploits solvatochromic fluorogenic nile red extract emission spectrum each dye molecule simultaneously enabling mapping...
Aggregation of α-synuclein leads to the formation oligomeric intermediates that can interact with membranes form pores. However, it is unknown how this cell toxicity in Parkinson's disease. We investigated species-specific effects on Ca(2+) signalling primary neurons and astrocytes using live neuronal imaging electrophysiology artificial membranes. demonstrate induces an increase basal intracellular its unfolded monomeric state as well state. Electrophysiology demonstrated monomers induce...
Abstract Aggregation of alpha-synuclein (α-Syn) drives Parkinson’s disease (PD), although the initial stages self-assembly and structural conversion have not been directly observed inside neurons. In this study, we tracked intracellular conformational states α-Syn using a single-molecule Förster resonance energy transfer (smFRET) biosensor, show here that converts from monomeric state into two distinct oligomeric in neurons concentration-dependent sequence-specific manner. Three-dimensional...
In Alzheimer's disease, fibrillar tau pathology accumulates and spreads through the brain synapses are lost. Evidence from mouse models indicates that trans-synaptically pre- to postsynapses oligomeric is synaptotoxic, but data on synaptic in human scarce. Here we used sub-diffraction-limit microscopy study accumulation postmortem temporal occipital cortices of control donors. Oligomeric present postsynaptic terminals, even areas without abundant deposition. Furthermore, there a higher...
Abstract α -Synuclein becomes misfolded and aggregated upon damage by various factors, for example, reactive oxygen species. These forms have been proposed to differential toxicities their interaction with mitochondria may cause dysfunction within this organelle that contributes the pathogenesis of Parkinson’s disease (PD). In particular, association -synuclein occurs through mitochondrial complex I importantly defects protein linked PD. Therefore, we investigated relationship between...
Abstract The molecular features of synapses in the hippocampus underpin current models learning and cognition. Although synapse ultra-structural diversity has been described canonical hippocampal circuitry, our knowledge sub-synaptic organisation synaptic molecules remains largely unknown. To address this, mice were engineered to express Post Synaptic Density 95 protein (PSD95) fused either eGFP or mEos2 imaged with two orthogonal super-resolution methods: gated stimulated emission depletion...
Abstract Parkinson's disease is the second most common neurodegenerative and its pathogenesis closely associated with oxidative stress. Deposition of aggregated α‐synuclein (α‐Syn) occurs in familial sporadic forms disease. Here, we studied effect oligomeric α‐Syn on one major markers stress, lipid peroxidation, primary co‐cultures neurons astrocytes. We found that but not monomeric significantly increases rate production reactive oxygen species, subsequently inducing peroxidation both...
The transport and trafficking of metabolites are critical for the correct functioning live cells. However, in situ metabolic imaging studies hampered by lack fluorescent chemical structures that allow direct monitoring small under physiological conditions with high spatial temporal resolution. Herein, we describe SCOTfluors as novel small-sized multi-colored fluorophores real-time tracking essential cells vivo acquisition profiles from human cancer variable origin.
As a key player of the protein quality control network cell, molecular chaperone Hsp70 inhibits aggregation amyloid tau. To date, mechanism this inhibition and tau species targeted by remain unknown. This is partly due to inherent difficulty studying aggregates because their heterogeneous transient nature. Here, we used ensemble single-molecule fluorescence measurements dissect how counteracts self-assembly process K18 ΔK280 variant. We found that blocks early stages suppressing formation...
The misfolding and aggregation of proteins into amyloid fibrils characterizes many neurodegenerative disorders such as Parkinson's Alzheimer's diseases. We report here a method, termed SAVE (single aggregate visualization by enhancement) imaging, for the ultrasensitive detection individual oligomers using single-molecule fluorescence microscopy. demonstrate that this method is able to detect presence aggregates α-synuclein, tau, amyloid-β. In addition, we show can also be identified in human...
Proteins fold into a single structural ensemble but can also misfold many diverse structures including small aggregates and fibrils, which differ in their toxicity. The aggregate surface properties play an important role how they interact with the plasma membrane cellular organelles, potentially inducing toxicity, however, these have not been measured to date due lack of suitable methods. Here, we used spectrally resolved, super-resolution imaging method combined environmentally sensitive...
Abstract Oligomers of alpha-synuclein are toxic to cells and have been proposed play a key role in the etiopathogenesis Parkinson’s disease. As certain missense mutations gene encoding for induce early-onset forms disease, it has suggested that these variants might an inherent tendency produce high concentrations oligomers during aggregation, although direct experimental evidence this is still missing. We used single-molecule Förster Resonance Energy Transfer visualize directly protein...
Abstract TDP-43 is an aggregation-prone protein which accumulates in the hallmark pathological inclusions of amyotrophic lateral sclerosis (ALS). However, analysis deeply phenotyped human post-mortem samples has shown that aggregation, revealed by standard antibody methods, correlates poorly with symptom manifestation. Recent identification cryptic-splicing events, such as detection Stathmin-2 ( STMN-2 ) cryptic exons, are providing evidence implicating loss-of-function a potential driving...
α-Synuclein oligomers can be toxic to cells and may responsible for cell death in Parkinson's disease. Their typically low abundance highly heterogeneous nature, however, make such species challenging study using traditional biochemical techniques. By combining fast-flow microfluidics with single-molecule fluorescence, we are able rapidly follow the process by which of αS formed characterize themselves. We have used technique show that populations different FRET efficiencies varying...
The aggregation of the protein ɑ-synuclein (ɑS) underlies a range increasingly common neurodegenerative disorders including Parkinson's disease. One widely explored therapeutic strategy for these conditions is use antibodies to target aggregated ɑS, although detailed molecular-level mechanism action such species remains elusive. Here, we characterize ɑS in vitro presence two ɑS-specific single-domain (nanobodies), NbSyn2 and NbSyn87, which bind highly accessible C-terminal region ɑS.We show...
The aberrant aggregation of α-synuclein is associated with several human diseases, collectively termed the α-synucleinopathies, which includes Parkinson's disease. progression these diseases is, in part, mediated by extracellular oligomers that may exert effects through mechanisms, including prion-like transfer, direct cytotoxicity, and pro-inflammatory actions. In this study, we show two abundant chaperones, clusterin α2-macroglobulin, directly bind to exposed hydrophobic regions on surface...
Significance The function of the T-cell coreceptor CD4 presents a long-standing puzzle. Although it is among most potent modulators immune responses, interacts with its binding partner, peptide-major histocompatibility class II (pMHC II), previously unmeasurably low affinity. Here, we set new upper limit for solution affinity and pMHC show that two-dimensional dissociation constant in supported lipid bilayers as much two to three orders magnitude higher than other interacting leukocyte...
We present LIVE-PAINT, a new approach to super-resolution fluorescent imaging inside live cells. In LIVE-PAINT only short peptide sequence is fused the protein being studied, unlike conventional methods, which rely on directly fusing biomolecule of interest large protein, organic fluorophore, or oligonucleotide. works by observing blinking localized fluorescence as this reversibly bound that protein. have demonstrated effectiveness number different proteins S. cerevisiae. Not widely...