Multiplex Immunofluorescent Analysis of Alpha-Synuclein in Nigral Lewy Bodies With Heat-Induced Antibody Stripping Reveals an Intricate Multilayered Structure

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00098892%3A_____%2F25%3A10159275" target="_blank" >RIV/00098892:_____/25:10159275 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61989592:15110/25:73632723

Výsledek na webu

<a href="https://onlinelibrary.wiley.com/doi/10.1111/nan.70024" target="_blank" >https://onlinelibrary.wiley.com/doi/10.1111/nan.70024</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1111/nan.70024" target="_blank" >10.1111/nan.70024</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Multiplex Immunofluorescent Analysis of Alpha-Synuclein in Nigral Lewy Bodies With Heat-Induced Antibody Stripping Reveals an Intricate Multilayered Structure

Popis výsledku v původním jazyce

Lewy bodies, the hallmark intracellular inclusions in Parkinson’s disease and dementia with Lewy bodies, exhibit complex architecture comprising multiple alpha-synuclein proteoforms, ubiquitin, cytoskeletal elements, and organelles. In this study, we applied a cost-effective and tissue-conserving multiplex immunofluorescence technique using heat-induced antibody stripping to examine the structural organization of nigral Lewy bodies in postmortem brain tissue from 11 patients with Lewy body disease. Using a panel of well-characterized alpha-synuclein antibodies targeting different domains, we analysed 71 brainstem Lewy bodies and identified a consistent concentric “onion-like” architecture. The C-terminal and N-terminal domains were predominantly localized to the outer shell, while the NAC domain displayed diffuse distribution. Notably, different alpha-synuclein epitopes showed variable colocalization, reflecting the influence of truncation and post-translational modifications on antibody binding. In contrast, pale bodies lacked clear structural organization. Our findings highlight the structural heterogeneity of Lewy bodies and support the use of multiple domain-specific antibodies to reliably identify and study these inclusions. This multiplex approach offers a scalable and accessible method for detailed spatial proteoform mapping, with potential applications in both research and neuropathological diagnostics.

Název v anglickém jazyce

Multiplex Immunofluorescent Analysis of Alpha-Synuclein in Nigral Lewy Bodies With Heat-Induced Antibody Stripping Reveals an Intricate Multilayered Structure

Popis výsledku anglicky

Lewy bodies, the hallmark intracellular inclusions in Parkinson’s disease and dementia with Lewy bodies, exhibit complex architecture comprising multiple alpha-synuclein proteoforms, ubiquitin, cytoskeletal elements, and organelles. In this study, we applied a cost-effective and tissue-conserving multiplex immunofluorescence technique using heat-induced antibody stripping to examine the structural organization of nigral Lewy bodies in postmortem brain tissue from 11 patients with Lewy body disease. Using a panel of well-characterized alpha-synuclein antibodies targeting different domains, we analysed 71 brainstem Lewy bodies and identified a consistent concentric “onion-like” architecture. The C-terminal and N-terminal domains were predominantly localized to the outer shell, while the NAC domain displayed diffuse distribution. Notably, different alpha-synuclein epitopes showed variable colocalization, reflecting the influence of truncation and post-translational modifications on antibody binding. In contrast, pale bodies lacked clear structural organization. Our findings highlight the structural heterogeneity of Lewy bodies and support the use of multiple domain-specific antibodies to reliably identify and study these inclusions. This multiplex approach offers a scalable and accessible method for detailed spatial proteoform mapping, with potential applications in both research and neuropathological diagnostics.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30109 - Pathology

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Neuropathology and Applied Neurobiology

ISSN

0305-1846

e-ISSN

1365-2990

Svazek periodika

51

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

5

Strana od-do

"e70024"

Kód UT WoS článku

001498975800001

EID výsledku v databázi Scopus

2-s2.0-105006853751