Project description:Lysosomes are cellular recycling stations and metabolic signaling hubs. Whether lysosome dynamics regulate mammalian brain development is unknown. We found that radial glia cells possess a large number of endolysosomes and that asymmetric inheritance of lysosomes in daughters of radial glia cells can predict fate and cell cycle length. To determine the lysosomal regulation of translation initiation by mTORC1/eIF4E axis, we performed RNA immunoprecipitation sequencing (RIP-seq) with antibody against eIF4E in E13.5 neocortex.
Project description:Lysosomes are cellular recycling stations and metabolic signaling hubs. Whether lysosome dynamics regulate mammalian brain development is unknown. We found that radial glia cells possess a large number of endolysosomes and that asymmetric inheritance of lysosomes in daughters of radial glia cells can predict fate and cell cycle length. To determine the translation control by the mTORC1, we performed polysome profiling of mRNAs associated with >3 ribosomes for neural progenitor cells that treated with DMSO or Torin1 for 2 hours.
Project description:Gene regulation in mammals involves a complex interplay between promoter and distal regulatory elements that function in concert to drive precise spatio-temporal gene expression programs. However, the dynamics of distal gene regulatory elements and its function in transcriptional reprogramming that underlies neurogenesis and neuronal activity remain largely unknown. Here we use a combinatorial analysis of genomewide datasets for chromatin accessibility (FAIRE-Seq) and enhancer mark H3K27ac to reveal a highly dynamic nature of chromatin accessibility during neurogenesis that gets restricted to certain genomic regions as neurons acquire a post-mitotic, terminally differentiated state. We further reveal that the distal open regions serve as target sites of distinct transcription factors that function in a stage-specific manner to contribute to the transcriptional program underlying neuronal commitment and maturation. A prolonged NMDA-driven neural activity results in epigenetic reprogramming at a large number of distal regulatory elements as well as dramatic reorganization of super-enhancers that in turn mediate critical transcriptional responses. Taken together, these findings reveal dynamics of distal regulatory landscape during neurogenesis and uncover novel regulatory elements that function in concert with epigenetic mechanisms and transcription factors to generate transcriptome underlying neuronal development and function. FAIRE-Seq and H3K27ac profiles for three stages on neuronal differentation viz. neuronal progenitors, day 1 neurons and day 10 neurons, were generated to understand the dynamics of accessible and ehancer chromatin landscape. Along with this we also generated RNASeq and H3K27ac profiles for day 10 neurons upon control and NMDA treatment.
Project description:Gene regulation in mammals involves a complex interplay between promoter and distal regulatory elements that function in concert to drive precise spatio-temporal gene expression programs. However, the dynamics of distal gene regulatory elements and its function in transcriptional reprogramming that underlies neurogenesis and neuronal activity remain largely unknown. Here we use a combinatorial analysis of genomewide datasets for chromatin accessibility (FAIRE-Seq) and enhancer mark H3K27ac to reveal a highly dynamic nature of chromatin accessibility during neurogenesis that gets restricted to certain genomic regions as neurons acquire a post-mitotic, terminally differentiated state. We further reveal that the distal open regions serve as target sites of distinct transcription factors that function in a stage-specific manner to contribute to the transcriptional program underlying neuronal commitment and maturation. A prolonged NMDA-driven neural activity results in epigenetic reprogramming at a large number of distal regulatory elements as well as dramatic reorganization of super-enhancers that in turn mediate critical transcriptional responses. Taken together, these findings reveal dynamics of distal regulatory landscape during neurogenesis and uncover novel regulatory elements that function in concert with epigenetic mechanisms and transcription factors to generate transcriptome underlying neuronal development and function.
Project description:During Drosophila melanogaster embryogenesis, a tight regulation of gene expression in time and space is required for the orderly emergence of the various specific cell types. While the general importance of microRNAs in modulating and regulating eukaryotic gene expression has already been well-established, their role in early neurogenesis remains to be addressed. In this survey, we investigate the transcriptional dynamics of microRNAs and their target transcripts during the neurogenesis of Drosophila melanogaster. To this end, we use the recently developed DIV-MARIS protocol, a method for enriching specific cell types from the Drosophila embryo in an in vivo setting, to sequence the tissue-specific transcriptomes. We generate dedicated small and total RNA-seq libraries for neuroblasts, neurons and glia cells at an early (6–8 h after egg laying) and late (18–22 h after egg laying) developmental stage. This strategy allows us to directly compare the transcriptomes of these cell types and investigate the potential functional roles of individual microRNAs with unprecedented spatiotemporal resolution, which is beyond the capabilities of existing in-situ hybridization studies. In total, we identify 74 microRNAs that are significantly differentially expressed between the three cell types and the two developmental stages.