Project description:The shift from glycolysis to oxidative phosphorylation is a key step in neural differentiation. Yet, the timing of metabolic rewiring in the development from progenitors to neurons remains elusive, especially in human corticogenesis. To bridge this gap, here we adopted a unique cell platform based on neocortex-derived human neuroepithelial stem cells, modeling their physiological transition into functional neurons in a four-month longitudinal study. Using quantitative proteomics, combined with NAD(P)H fluorescence lifetime imaging in living cells, we investigated the entire differentiation process and described how metabolic pathways drive cortical neuron maturation. The prevalent metabolic adaptations were then confirmed in cerebro-cortical organoids and in a human neocortical specimen. This study provides a spatio-temporal map of neocortical metabolism during development and offers a framework to investigate human neurometabolic disorders.
Project description:Affymetrix Mouse Genome 430 2.0 GeneChip microarrays were used to analyze murine neocortical and cerbellar astrocytes generated from postnatal (PN) day 1 wild-type (ICR) pups. Experiment Overall Design: Three samples each of murine neocortical and cerebellar astrocytes were analyzed.
Project description:Affymetrix Mouse Genome 430 2.0 GeneChip microarrays were used to analyze murine neocortical and cerbellar astrocytes generated from postnatal (PN) day 1 wild-type (ICR) pups. Keywords: neocortical astrocyte, cerebellar astrocyte, murine, postnatal day 1
Project description:To investigate BRN1/2 function in neocortical development, we knockout BRN1/2 in the dorsal telencephalon (BRN1/2 cKO) We then performed gene expression profiling analysis using data obtained from scRNA-seq of 13 different mice at E12.5 and E14.5
Project description:Precise spatiotemporal control of mRNA translation machinery is essential to proper development of highly complex systems like the neocortex. Here, we show that an RNA-binding protein, Hu antigen R (HuR), regulates both neocorticogenesis and specificity of neocortical translation machinery in a developmental stagedependent manner in mice. Neocortical absence of HuR alters the phosphorylation states of the initiation and elongation factors of the core translation machinery. In addition, HuR regulates the temporally specific positioning of functionally related mRNAs into the active translation sites, the polysomes. HuR also determines the specificity of neocortical polysomes by defining their combinatorial composition of ribosomal proteins and initiation and elongation factors. For some of the HuR-dependent proteins, the association with polysomes depends on the eIF2 alpha kinase 4 (eIF2ak4), which associated with HuR in prenatal developing neocortices. Finally, we found that deletion of HuR prior to embryonic day 10 (E10) disrupts both neocortical lamination and formation of the main neocortical commissure, the corpus callosum. Our study identifies a crucial role for HuR in neocortical development as a translational gatekeeper for functionally related mRNA subgroups and polysomal protein specificity. Cortex was dissected from mouse pups at embryonic day 13 (E13) or the day of birth (P0).
Project description:Transcriptional changes of mouse precision-cut liver slices (PCLS) after three days of culture were determined using RNA sequencing. PCLS were cultured for three days in the absence or presence of 2.5 mM valproic acid sodium salt (VPA). Illumina NovaSeq SP was used for sequencing.
