A number of neurodegenerative conditions are associated with the formation of cytosolic inclusions of TDP-43 within neurons. We expressed full-length TDP-43 in a motoneuron/neuroblastoma hybrid cell line (NSC-34) and exploited the high-resolution power of stimulated emission depletion microscopy to monitor the changes of nuclear and cytoplasmic TDP-43 levels and the formation of various size classes of cytoplasmic TDP-43 aggregates with time. Concomitantly, we monitored oxidative stress and mitochondrial impairment using the MitoSOX and MTT reduction assays, respectively. Using a quantitative biology approach, we attributed neuronal dysfunction associated with cytoplasmic deposition component to the formation of the largest inclusions, independently of stress granules. This is in contrast to other neurodegenerative diseases where toxicity is attributed to small oligomers. Using specific inhibitors, markers, and electron microscopy, the proteasome and autophagy were found to target mainly the largest deleterious inclusions, but their efficiency soon decreases without full recovery of neuronal viability.

A quantitative biology approach correlates neuronal toxicity with the largest inclusions of TDP-43 / Roberta Cascella, Alessandra Bigi, Dylan Giorgino Riffert, Maria Cristina Gagliani, Emilio Ermini, Matteo Moretti, Katia Cortese, Cristina Cecchi, Fabrizio Chiti. - In: SCIENCE ADVANCES. - ISSN 2375-2548. - ELETTRONICO. - eabm6376:(2022), pp. 1-13. [10.1126/sciadv.abm6376]

A quantitative biology approach correlates neuronal toxicity with the largest inclusions of TDP-43

Roberta Cascella;Alessandra Bigi;Matteo Moretti;Cristina Cecchi
;
Fabrizio Chiti
2022

Abstract

A number of neurodegenerative conditions are associated with the formation of cytosolic inclusions of TDP-43 within neurons. We expressed full-length TDP-43 in a motoneuron/neuroblastoma hybrid cell line (NSC-34) and exploited the high-resolution power of stimulated emission depletion microscopy to monitor the changes of nuclear and cytoplasmic TDP-43 levels and the formation of various size classes of cytoplasmic TDP-43 aggregates with time. Concomitantly, we monitored oxidative stress and mitochondrial impairment using the MitoSOX and MTT reduction assays, respectively. Using a quantitative biology approach, we attributed neuronal dysfunction associated with cytoplasmic deposition component to the formation of the largest inclusions, independently of stress granules. This is in contrast to other neurodegenerative diseases where toxicity is attributed to small oligomers. Using specific inhibitors, markers, and electron microscopy, the proteasome and autophagy were found to target mainly the largest deleterious inclusions, but their efficiency soon decreases without full recovery of neuronal viability.
2022
eabm6376
1
13
Roberta Cascella, Alessandra Bigi, Dylan Giorgino Riffert, Maria Cristina Gagliani, Emilio Ermini, Matteo Moretti, Katia Cortese, Cristina Cecchi, Fabrizio Chiti
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/1281049
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