STochastic Optical Reconstruction Microscopy (STORM) reveals the nanoscale organization of pathological aggregates in human brain

Abstract

Optical Reconstruction Microscopy (STORM) reveals the nanoscale organization of pathological aggregates in human brain Abstract: Aims: Histological analysis of brain tissue samples provides valuable information about the pathological processes leading to common neurodegenerative disorders. In this context, the development of novel high-resolution imaging approaches is a current challenge in neuroscience. Methods: To this end, we used a recent super-resolution imaging technique called STochastic Optical Reconstruction Microscopy (STORM) to analyse human brain sections. We combined STORM cell imaging protocols with neuropathological techniques to image cryopreserved brain samples from control subjects and patients with neurodegenerative diseases. Results: This approach allowed us to perform 2D-, 3D-and two-colour-STORM in neocortex, white matter and brainstem samples. STORM proved to be particularly effective at visualizing the organization of dense protein inclusions and we imaged with a <50 nm resolution pathological aggregates within the central nervous system of patients with Alzheimer's disease, Parkinson's disease, Lewy body dementia and fronto-temporal lobar degeneration. Aggregated Ab branches appeared reticulated and cross-linked in the extracellular matrix, with widths from 60 to 240 nm. Intraneuronal Tau and TDP-43 inclusions were denser, with a honeycomb pattern in the soma and a filamentous organization in the axons. Finally, STORM imaging of a-synuclein pathology revealed the internal organization of Lewy bodies that could not be observed by conventional fluorescence microscopy. Conclusions: STORM imaging of human brain samples opens further gates to a more comprehensive understanding of common neurological disorders. The convenience of this technique should open a straightforward extension of its application for super-resolution imaging of the human brain, with promising avenues to current challenges in neuroscience.

The subcellular arrangement of alpha-synuclein proteoforms in the Parkinson’s disease brain as revealed by multicolor STED microscopy

“…Distribution patterns for different aSyn epitopes were also analyzed in neuronal inclusions without ring-shaped appearance, in the same sections (in case of SN) and in other sections of the same patients (hippocampus/transentorhinal cortex). Although unambiguously detected and outlined by uniform Ser129-p aSyn immunolabeling—with exception of typical ‘vacuolar-like structures’ lacking Ser129-p aSyn reactivity [ 8 , 67 ]—these inclusions often revealed weaker signal intensities for the other epitopes (particularly 122CTT, NAC and NT aSyn) that were diffusely distributed throughout the inclusion (Online Resource Fig. 3a,d).…”

Optical Microscopy and the Extracellular Matrix Structure: A Review

“…They demonstrated that integrins exist as a patterned array on the surface of neutrophils and showed that STORM is a viable technique in the exploration of molecule patterning in cell–matrix interactions [ 88 ]. Codron et al used STORM to reveal the organisation of aggregates in the brain [ 92 ]. Aggregates in the brain are one of the hallmarks of Alzheimer’s disease, formed by extracellular deposits of Amyloid-β peptides (Aβ) and interneuronal aggregates of hyperphosphorylated tau protein (p.Tau) [ 92 ].…”

“…Codron et al used STORM to reveal the organisation of aggregates in the brain [ 92 ]. Aggregates in the brain are one of the hallmarks of Alzheimer’s disease, formed by extracellular deposits of Amyloid-β peptides (Aβ) and interneuronal aggregates of hyperphosphorylated tau protein (p.Tau) [ 92 ]. Codron et al used STORM to explore the structure and organization of extracellular deposits by targeting Aβ and p.Tau, demonstrating the use of the super-fluorescent technique in observation of the structure of pathological structures within the brain [ 92 ].…”

Molecular Imaging of Tau Protein: New Insights and Future Directions

“…While the density of exogenous tau fibrils binding to neurons increased over time, the radius of clusters was not affected (Shrivastava et al, 2019). STORM has been combined with neuropathological immunohistochemistry methods to image ptau in the prefrontal, parietal, and temporal cortex of AD patients with a resolution <50 nm (Codron et al, 2020). Tau aggregates appeared organized in filamentous structures at the axonal level while at the soma, they presented a honeycombed structure most likely arising from the presence of proteins and organelles.…”

“…Note the difference in resolution between image in panels (A2) and (B2). Modified fromCodron et al (2020) under the Creative Commons CC-BY-NC-ND.…”