eprintid: 10162398
rev_number: 7
eprint_status: archive
userid: 699
dir: disk0/10/16/23/98
datestamp: 2023-01-05 12:49:29
lastmod: 2023-01-05 12:49:29
status_changed: 2023-01-05 12:49:29
type: article
metadata_visibility: show
sword_depositor: 699
creators_name: Croft, Cara L
creators_name: Paterno, Giavanna
creators_name: Vause, Ava R
creators_name: Rowe, Lyla A
creators_name: Ryu, Daniel H
creators_name: Goodwin, Marshall S
creators_name: Moran, Corey A
creators_name: Cruz, Pedro E
creators_name: Giasson, Benoit I
creators_name: Golde, Todd E
title: Optical pulse labeling studies reveal exogenous seeding slows α-synuclein clearance
ispublished: pub
divisions: UCL
divisions: B02
divisions: C07
divisions: D07
divisions: FD9
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abstract: The accumulation of α-synuclein (α-syn) in intracellular formations known as Lewy bodies (LBs) is associated with several neurodegenerative diseases including Parkinson's disease and Lewy Body Dementia. There is still limited understanding of how α-syn and LB formation is associated with cellular dysfunction and degeneration in these diseases. To examine the clearance and production dynamics of α-syn we transduced organotypic murine brain slice cultures (BSCs) with recombinant adeno-associated viruses (rAAVs) to express Dendra2-tagged human wild-type (WT) and mutant A53T α-syn, with and without the addition of exogenous α-syn fibrillar seeds and tracked them over several weeks in culture using optical pulse labeling. We found that neurons expressing WT or mutant A53T human α-syn show similar rates of α-syn turnover even when insoluble, phosphorylated Ser129 α-syn has accumulated. Taken together, this data reveals α-syn aggregation and overexpression, pSer129 α-syn, nor the A53T mutation affect α-syn dynamics in this system. Prion-type seeding with exogenous α-syn fibrils significantly slows α-syn turnover, in the absence of toxicity but is associated with the accumulation of anti-p62 immunoreactivity and Thiazin Red positivity. Prion-type induction of α-syn aggregation points towards a potential protein clearance deficit in the presence of fibrillar seeds and the ease of this system to explore precise mechanisms underlying these processes. This system facilitates the exploration of α-syn protein dynamics over long-term culture periods. This platform can further be exploited to provide mechanistic insight on what drives this slowing of α-syn turnover and how therapeutics, other genes or different α-syn mutations may affect α-syn protein dynamics.
date: 2022-12-19
date_type: published
publisher: Springer Science and Business Media LLC
official_url: https://doi.org/10.1038/s41531-022-00434-4
oa_status: green
full_text_type: pub
language: eng
primo: open
primo_central: open_green
verified: verified_manual
elements_id: 1997264
doi: 10.1038/s41531-022-00434-4
medium: Electronic
pii: 10.1038/s41531-022-00434-4
lyricists_name: Croft, Cara
lyricists_id: CCROF38
actors_name: Flynn, Bernadette
actors_id: BFFLY94
actors_role: owner
funding_acknowledgements: U01TR003715 [U.S. Department of Health & Human Services | NIH | National Institute on Aging (U.S. National Institute on Aging)]; U01AG046139 [U.S. Department of Health & Human Services | NIH | National Institute on Aging (U.S. National Institute on Aging)]; ARUK-RADF2019A-003 [Alzheimer's Research UK (ARUK)]
full_text_status: public
publication: npj Parkinson's Disease
volume: 8
article_number: 173
event_location: United States
citation:        Croft, Cara L;    Paterno, Giavanna;    Vause, Ava R;    Rowe, Lyla A;    Ryu, Daniel H;    Goodwin, Marshall S;    Moran, Corey A;             ... Golde, Todd E; + view all <#>        Croft, Cara L;  Paterno, Giavanna;  Vause, Ava R;  Rowe, Lyla A;  Ryu, Daniel H;  Goodwin, Marshall S;  Moran, Corey A;  Cruz, Pedro E;  Giasson, Benoit I;  Golde, Todd E;   - view fewer <#>    (2022)    Optical pulse labeling studies reveal exogenous seeding slows α-synuclein clearance.                   npj Parkinson's Disease , 8     , Article 173.  10.1038/s41531-022-00434-4 <https://doi.org/10.1038/s41531-022-00434-4>.       Green open access   
 
document_url: https://discovery.ucl.ac.uk/id/eprint/10162398/1/s41531-022-00434-4.pdf