Notch directs telencephalic development and neuron fate determination by regulating miRNA levels

The central nervous system (CNS) contains myriads of different types of cells produced from multipotent neural progenitors through multiple rounds of cell divisions. Neural progenitors acquire distinct cell identities depending on their spatial position, but they are also influenced by temporal cues to give rise to different cell populations over time. For instance, the progenitors of the cerebral neocortex generate different populations of excitatory projection neurons following a well-known sequence that is conserved across species. The Notch signaling pathway plays crucial roles in CNS development, including regulating the balance between proliferation and differentiation and fate acquisition. However, the molecular mechanisms by which Notch impacts progenitor fate decisions and telencephalic patterning have not been fully resolved. Here, we show that Notch signaling is essential for neocortical and hippocampal morphogenesis as well as for the development of the corpus callosum an...,

FlashTag NSC labelling and FACS
Labelling of cortical neuronal progenitors with carboxyfluorescein esters (CFSEs) was achieved as described elsewhere (Govindan et al., 2018). Briefly, 1μl of a 5mM solution of CellTrace CFSE (from the CellTrace CFSE Cell Proliferation Kit, Invitrogen #C34554) and 0.01% FastGreen in DMSO was injected into the 3rd ventricle of E13.5 control and NICD embryos. Dams were allowed to recover and injected embryos were collected 1 hour post-injection. Embryonic cortices were dissected individually and dissociated into single cells using Papain Dissociation System (Worthington, Cat# LK003150) using manufacturer’s protocol. Cells were resuspended in FACS media (DMEM/F12 without phenol red supplemented with 10% FBS and B-27) and sorted using a Beckman Coulter Astrios EQ Cell Sorter.
RNA and miRNA sequencing
Total RNA from control (n=3) and NICD (n=5) sorted cells was extracted using the Total RNA Purification Plus Kit (NORGEN Biotek Corp., #48300). Gene express..., There are two sets of files: 1) Excel spreadsheet files and web browser files can be opened with Excel (Microsoft) software and any web browser; 2) Fastq sequencing files can be open with R. R is available as free software.