RRC ID 61358
Author Ishii T, Kawakami E, Endo K, Misawa H, Watabe K.
Title Formation and spreading of TDP-43 aggregates in cultured neuronal and glial cells demonstrated by time-lapse imaging.
Journal PLoS One
Abstract TAR DNA-binding protein 43 (TDP-43) is a main constituent of cytoplasmic aggregates in neuronal and glial cells in cases of amyotrophic lateral sclerosis and frontotemporal lobar degeneration. We have previously demonstrated that adenovirus-transduced artificial TDP-43 cytoplasmic aggregates formation is enhanced by proteasome inhibition in vitro and in vivo. However, the relationship between cytoplasmic aggregate formation and cell death remains unclear. In the present study, rat neural stem cell lines stably transfected with EGFP- or Sirius-expression vectors under the control of tubulin beta III, glial fibrillary acidic protein, or 2',3'-cyclic nucleotide 3'-phosphodiesterase promoter were differentiated into neurons, astrocytes, and oligodendrocytes, respectively, in the presence of retinoic acid. The differentiated cells were then transduced with adenoviruses expressing DsRed-tagged human wild type and C-terminal fragment TDP-43 under the condition of proteasome inhibition. Time-lapse imaging analyses revealed growing cytoplasmic aggregates in the transduced neuronal and glial cells, followed by collapse of the cell. The aggregates remained insoluble in culture media, consisted of sarkosyl-insoluble granular materials, and contained phosphorylated TDP-43. Moreover, the released aggregates were incorporated into neighboring neuronal cells, suggesting cell-to-cell spreading. The present study provides a novel tool for analyzing the detailed molecular mechanisms of TDP-43 proteinopathy in vitro.
Volume 12(6)
Pages e0179375
Published 2017-1-1
DOI 10.1371/journal.pone.0179375
PII PONE-D-16-44831
PMID 28599005
PMC PMC5466347
MeSH Animals Cell Death Cells, Cultured DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism* Humans Neural Stem Cells / metabolism Neural Stem Cells / pathology Neural Stem Cells / ultrastructure Neuroglia / metabolism* Neuroglia / pathology Neuroglia / ultrastructure Neurons / metabolism* Neurons / pathology Neurons / ultrastructure Phosphorylation Proteasome Endopeptidase Complex / metabolism Protein Aggregates* Protein Aggregation, Pathological* Protein Transport RNA, Small Interfering / genetics Rats Time-Lapse Imaging
IF 2.74
DNA material AxDsR-WT.TDP43 (RDB15499) AxDsR-CTF.TDP43 (RDB15500)