RRC ID 61359
Author Watabe K, Akiyama K, Kawakami E, Ishii T, Endo K, Yanagisawa H, Sango K, Tsukamoto M.
Title Adenoviral expression of TDP-43 and FUS genes and shRNAs for protein degradation pathways in rodent motoneurons in vitro and in vivo.
Journal Neuropathology
Abstract Formation of cytoplasmic aggregates in neuronal and glial cells is one of the pathological hallmarks of amyotrophic lateral sclerosis (ALS). Mutations in two genes encoding transactivation response (TAR) DNA-binding protein 43 (TDP-43) and fused in sarcoma (FUS), both of which are main constituents of cytoplasmic aggregates, have been identified in patients with familial and sporadic ALS. Impairment of protein degradation machineries has also been recognized to participate in motoneuron degeneration in ALS. In the present study, we produced recombinant adenovirus vectors encoding wild type and mutant TDP-43 and FUS, and those encoding short hairpin RNAs (shRNAs) for proteasome (PSMC1), autophagy (ATG5), and endosome (VPS24) systems to investigate whether the coupled gene transductions in motoneurons by these adenoviruses elicit ALS pathology. Cultured neurons, astrocytes and oligodendrocytes differentiated from adult rat neural stem cells and motoneurons derived from mouse embryonic stem cells were successfully infected with these adenoviruses showing cytoplasmic aggregate formation. When these adenoviruses were injected into the facial nerves of adult rats, exogenous TDP-43 and FUS proteins were strongly expressed in facial motoneurons by a retrograde axonal transport of the adenoviruses. Co-infections of adenovirus encoding shRNA for PSMC1, ATG5 or VPS24 with TDP-43 or FUS adenovirus enhanced cytoplasmic aggregate formation in facial motoneurons, suggesting that impairment of protein degradation pathways accelerates formation of TDP-43 and FUS-positive aggregates in ALS.
Volume 34(1)
Pages 83-98
Published 2014-2-1
DOI 10.1111/neup.12058
PMID 23937386
MeSH Adenoviridae / genetics Animals Astrocytes / metabolism Astrocytes / pathology Cells, Cultured DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism* Embryonic Stem Cells / metabolism Embryonic Stem Cells / pathology HEK293 Cells Humans Inclusion Bodies / metabolism Inclusion Bodies / ultrastructure* Male Mice Motor Neurons / metabolism Motor Neurons / ultrastructure* Oligodendroglia / metabolism Oligodendroglia / pathology RNA, Small Interfering / genetics RNA, Small Interfering / metabolism* RNA-Binding Protein FUS / genetics RNA-Binding Protein FUS / metabolism* Rats Rats, Inbred F344 Rats, Mutant Strains
IF 1.758
DNA material AxDsR-WT.TDP43 (RDB15499) AxDsR-CTF.TDP43 (RDB15500)