Journal
FRONTIERS IN MOLECULAR NEUROSCIENCE
Volume 13, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fnmol.2020.00046
Keywords
reprogrammed human neurons; nucleocytoplasmic transport; nuclear mRNA export; fluorescence in situ hybridization (FISH); protein nuclear transport; sex as a biological variable (SABV)
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Funding
- Friends of the Alzheimer's Disease Center (ADC) [NIH/NIA P30-12300-21]
- NIH ADC grant [NIH/NIA P30-12300-21]
- DoD PRMRP grant [W81XWH2010186]
- Welch Foundation [I-1724]
- Decherd Foundation
- Pape Adams Foundation
- Kent Waldrep Foundation Center for Basic Research on Nerve Growth and Regeneration
- NIH [NS099073, NS092616, NS088095, R21NS112910]
- U.S. Department of Defense (DOD) [W81XWH2010186] Funding Source: U.S. Department of Defense (DOD)
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Nucleocytoplasmic transport (NCT) across thenuclear envelope (NE) is tightly regulated in eukaryotic cells and iscritical for maintaining cellular homeostasis. Its dysregulationleads to aging and neurodegeneration. Because they maintainaging-associated hallmarks, directly reprogrammed neurons from human fibroblasts are invaluable in understanding NCT. However, it is not clear whether NCT activity is influenced by neuronal maturation and sample sex [a key biological variable emphasized by the National Institutes of Health (NIH) policy]. We examined here NCT activity at the single-cell level by measuring mRNA subcellular distribution and protein transport in directly induced motor neurons (diMNs) from adult human fibroblasts. The results show that mRNA subcellular distribution but not protein transport is affected by neuronal maturation stages, whereas both transport processes are not influenced by the sample sex. This study also provides quantitative methods and optimized conditions for measuring NCTs of mRNAs or protein cargoes, establishing a robust way for future functional examinations of NCT activity in directly induced neurons from diseased human patients.
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