EEF1A2 knockdown impairs neuronal proliferation and inhibits neurite outgrowth of differentiating neurons
Issued Date
2022-05-18
Resource Type
ISSN
09594965
eISSN
1473558X
Scopus ID
2-s2.0-85130862103
Pubmed ID
35594436
Journal Title
NeuroReport
Volume
33
Issue
8
Start Page
336
End Page
344
Rights Holder(s)
SCOPUS
Bibliographic Citation
NeuroReport Vol.33 No.8 (2022) , 336-344
Suggested Citation
Rumpansuwon K. EEF1A2 knockdown impairs neuronal proliferation and inhibits neurite outgrowth of differentiating neurons. NeuroReport Vol.33 No.8 (2022) , 336-344. 344. doi:10.1097/WNR.0000000000001791 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/86717
Title
EEF1A2 knockdown impairs neuronal proliferation and inhibits neurite outgrowth of differentiating neurons
Author(s)
Author's Affiliation
Other Contributor(s)
Abstract
Objectives The translation elongation factor-1, alpha-2 (eEF1A2) plays an important role in protein synthesis. Mutations in this gene have been described in individuals with neurodevelopmental disorders. Here, we silenced the expression of eEFA2 in human SH-SY5Y neuroblastoma cells and observed its roles in neuronal proliferation and differentiation upon induction with retinoic acid. Methods eEF1A2 were silenced using siRNA transfection. Cell proliferation was qualitatively evaluated by Ki-67 immunocytochemistry. Neuronal differentiation was induced with retinoic acid for 3, 5, 7 and 10 days. Neurite length was measured. The expression of microtubule-associated protein 2 (MAP2) was analyzed by western blotting. Tyrosine hydroxylase expression was visualized by immunofluorescence. Cytotoxicity to a neurotoxin, 1-methyl-4-phenylpyridinium (MPP+), was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and western blotting of cleaved caspase-3. Results eEF1A2 knockdown suppressed the proliferative activity of undifferentiated SH-SY5Y cells as shown by decreased Ki-67 immunostaining. Upon retinoic acid-induction, differentiated neurons with eEF1A2 knockdown exhibited shorter neurite length than untransfected cells, which was associated with the reduction of tyrosine hydroxylase and suppression of MAP2 at 10 days of differentiation. eEF1A2 knockdown decreased the survival of neurons, which was clearly observed in undifferentiated and short-term differentiated cells. Upon treatment with MPP+, cells with eEF1A2 knockdown showed a further reduction in cell survival and an increase of cleaved caspase-3 protein. Conclusions Our results suggest that eEF1A2 may be required for neuronal proliferation and differentiation of SH-SY5Y cells. Increased cell death susceptibility against MPP+in eEF1A2-knockdown neurons may imply the neuroprotective role of eEF1A2.