Byline: Kazuhide. Asakawa, Hiroshi. Handa, Koichi. Kawakami
Cytoplasmic inclusions containing the transactivation response element (TAR) DNA-binding protein-43 (TDP-43) aggregates are hallmarks of neurodegenerative disorders, such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (Arai et al., 2006; Neumann et al., 2006). Despite the well-recognized correlation between TDP-43 aggregation and neuronal degeneration, whether this relationship is causal has remained unclear. The recent advent of the optoDroplet technique for controlling protein-protein interaction through light illumination has allowed the generation of droplets containing intrinsically disordered proteins in cells with an unprecedented spatiotemporal precision (Shin et al., 2017). Moreover, the use of this optogenetic approach to explore TDP-43 uncovered the neurotoxicity associated with TDP-43 phase transitions in cultured neurons (Mann et al., 2019; Zhang et al., 2019). Here, we discuss our recent discovery of novel facets of TDP-43, based on the use of an optogenetic TDP-43 variant (opTDP-43) interrogated in zebrafish motor neurons, in which the in vivo dynamic nuclear-cytoplasmic relocation and the clustering of TDP-43 can be observed directly due to the transparent zebrafish body (Asakawa et al., 2020). Our results showed that optogenetically clumped opTDP-43 mislocalizes to the cytoplasm and damages motor neurons before the development of large cytoplasmic aggregates, which are similar to those found in the ALS patients. This unexpected finding raises the possibility that the onset of motor neuron dysfunction caused by TDP-43 in ALS occurs much earlier than previously anticipated; therefore, future efforts should be made to identify the cellular environments and insults that facilitate pathological TDP-43 oligomer formation to better understand, and potentially intervene in, the prodromal phase of ALS and other TDP-43 proteinopathies.
ALS is a devastating neurological disease, in which the upper and lower motor neurons progressively degenerate, leading to fatal paralysis due to relentless muscular atrophy. A characteristic feature of degenerating motor neurons in sporadic ALS, which accounts for greater than 90% of total ALS cases, is the deposition of an aggregated form of TDP-43 in the cytoplasm. The remaining 10% cases are associated with monogenic heritability (familial ALS), and approximately 4% of familial ALS cases have been linked to the TARDBP gene, which encodes TDP-43, demonstrating a causal role for TDP-43 in the pathogenesis of some forms ALS. At present, however, whether the etiological mechanisms of sporadic ALS, where TARDBP /TDP-43 mutation is absent, are shared with the TARDBP-linked familial ALS remains known.
TDP-43 is an evolutionarily conserved heterogeneous nuclear ribonucleoprotein that regulates various aspects of RNA metabolisms including transcription, splicing, and mRNA stability and transport. Under normal physiological conditions, TDP-43 is primarily localized in the nucleus, where it forms homo-oligomers through its N-terminal domain [Figure 1]A and [Figure 1]B (Afroz et al., 2017). On the other hand, pathological TDP-43 aggregation is prominent in the cytoplasm and mediated by the C-terminal intrinsically disordered region containing prion-like glutamine/asparagine-rich and glycine-rich regions, suggesting that the modes of TDP-43 multimerization may influence its subcellular localization, or vice versa . In addition, cytoplasmic degradation of TDP-43 protein by the ubiquitin-proteasome system and autophagy also contributes to the...