γ-secretase inhibitor disturbs the morphological development of differentiating neurons through affecting Notch/miR-342-5p

During neurodevelopment, differentiation of neural stem/progenitor cells (NSPCs) into neurons are regulated by many factors including Notch signaling pathway. Herein, we report the effect of a Notch signaling blocker, i.e. gamma-secretase inhibitor (GSI), on this differentiating process, especially on the morphological development. NSPCs were cultured and induced to differentiate with or without GSI. The neurite outgrowth was impeded by GSI application and the expression of a Notch signaling downstream effector miR-342-5p increased with the downregulated expression of Notch effectors Hes1 and Hes5. Upregulated expression of miR-342-5p in differentiating NSPCs could shorten the neurite length of progeny neurons, which was similar to the effect of GSI. To avoid the possible influence from astrocytes into neurons, we directly applied cultured neurons, on which GSI could shorten the processes and RBP-J knockdown could also reduce the neurite length. Similarly, transfection of miR-342-5p mimics or inhibitors into PC12 cells led to shorter or longer processes of cells compared with control ones. Furthermore, in differentiating NSPCs, GSI-induced shorter neurites could be partially rescued by miR-3425p inhibitors, and STAT3 was one of the possible targets of miR-342-5p during this differentiating process as indicated by results of Western Blot test, luciferase reporter assay and GFP reporter assay. To further demonstrate the role of STAT3, it was introduced into GSI-treated neurons and the GSI-affected neurites could also be partially rescued. In conclusion, GSI could influence the morphological development of neurons and the possible mechanism involved Notch/miR-342-5p and STAT3. These results would be informative for future therapeutic research.

Automated summary

During neurodevelopment, the differentiation of neural stem/progenitor cells into neurons is regulated by the Notch signaling pathway. In this study, the effect of a Notch signaling blocker, gamma-secretase inhibitor (GSI), on the morphological development of differentiating neurons was investigated. The results showed that GSI application impeded neurite outgrowth and increased the expression of miR-342-5p, a downstream effector of Notch signaling. Furthermore, the shorter neurites induced by GSI could be partially rescued by miR-342-5p inhibitors, indicating the involvement of STAT3 as a possible target of miR-342-5p in this process. These findings provide important insights for future therapeutic research.