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      Exploring the Complexity of Cortical Development Using Single-Cell Transcriptomics

      review-article
      ,
      Frontiers in Neuroscience
      Frontiers Media S.A.
      epigenetics, neurogenesis, development, neocortex, stem cells

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          Abstract

          The developing neocortex in the mammalian brain is composed of multiple cell types including apical progenitors (AP), basal progenitors (BP), and neurons that populate three different layers, the ventricular zone (VZ), the subventricular zone (SVZ), and the cortical plate (CP). Despite recent advances, the diversity of the existing cell populations including those which are differentiating and mature, their biogenesis and the underlying gene regulatory mechanisms remain poorly known. Recent studies have taken advantage of the rapidly emerging single-cell technologies to decode the heterogeneity of cell populations at the transcriptome level during cortical development and their molecular details. Here we review these studies and provide an overview of the steps in single-cell transcriptomics including both experimental and computational analysis. We also discuss how single-cell genomics holds a big potential in future for brain research and discuss its possible applications and biological insights that can be achieved from these approaches. We conclude this review by discussing the current challenges in the implementation of single-cell techniques toward a comprehensive understanding of the genetic and epigenetic mechanisms underlying neocortex development.

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          Most cited references36

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          The cell biology of neurogenesis.

          During the development of the mammalian central nervous system, neural stem cells and their derivative progenitor cells generate neurons by asymmetric and symmetric divisions. The proliferation versus differentiation of these cells and the type of division are closely linked to their epithelial characteristics, notably, their apical-basal polarity and cell-cycle length. Here, we discuss how these features change during development from neuroepithelial to radial glial cells, and how this transition affects cell fate and neurogenesis.
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            Comparative Analysis of Single-Cell RNA Sequencing Methods.

            Single-cell RNA sequencing (scRNA-seq) offers new possibilities to address biological and medical questions. However, systematic comparisons of the performance of diverse scRNA-seq protocols are lacking. We generated data from 583 mouse embryonic stem cells to evaluate six prominent scRNA-seq methods: CEL-seq2, Drop-seq, MARS-seq, SCRB-seq, Smart-seq, and Smart-seq2. While Smart-seq2 detected the most genes per cell and across cells, CEL-seq2, Drop-seq, MARS-seq, and SCRB-seq quantified mRNA levels with less amplification noise due to the use of unique molecular identifiers (UMIs). Power simulations at different sequencing depths showed that Drop-seq is more cost-efficient for transcriptome quantification of large numbers of cells, while MARS-seq, SCRB-seq, and Smart-seq2 are more efficient when analyzing fewer cells. Our quantitative comparison offers the basis for an informed choice among six prominent scRNA-seq methods, and it provides a framework for benchmarking further improvements of scRNA-seq protocols.
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              Single-Cell RNA-Seq with Waterfall Reveals Molecular Cascades underlying Adult Neurogenesis.

              Somatic stem cells contribute to tissue ontogenesis, homeostasis, and regeneration through sequential processes. Systematic molecular analysis of stem cell behavior is challenging because classic approaches cannot resolve cellular heterogeneity or capture developmental dynamics. Here we provide a comprehensive resource of single-cell transcriptomes of adult hippocampal quiescent neural stem cells (qNSCs) and their immediate progeny. We further developed Waterfall, a bioinformatic pipeline, to statistically quantify singe-cell gene expression along a de novo reconstructed continuous developmental trajectory. Our study reveals molecular signatures of adult qNSCs, characterized by active niche signaling integration and low protein translation capacity. Our analyses further delineate molecular cascades underlying qNSC activation and neurogenesis initiation, exemplified by decreased extrinsic signaling capacity, primed translational machinery, and regulatory switches in transcription factors, metabolism, and energy sources. Our study reveals the molecular continuum underlying adult neurogenesis and illustrates how Waterfall can be used for single-cell omics analyses of various continuous biological processes.
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                Author and article information

                Contributors
                Journal
                Front Neurosci
                Front Neurosci
                Front. Neurosci.
                Frontiers in Neuroscience
                Frontiers Media S.A.
                1662-4548
                1662-453X
                02 February 2018
                2018
                : 12
                : 31
                Affiliations
                Institute of Molecular Biology , Mainz, Germany
                Author notes

                Edited by: Mareike Albert, Max Planck Institute of Molecular Cell Biology and Genetics (MPG), Germany

                Reviewed by: Marta Florio, Harvard Medical School, United States; Madeline A. Lancaster, MRC Laboratory of Molecular Biology (MRC), United Kingdom

                *Correspondence: Vijay K. Tiwari v.tiwari@ 123456imb-mainz.de

                This article was submitted to Neurogenesis, a section of the journal Frontiers in Neuroscience

                †Present Address: Hyobin Jeong, Genome Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany

                Article
                10.3389/fnins.2018.00031
                5801402
                29456488
                e0a07b1e-4e6f-49b4-93ca-2315fe144ff0
                Copyright © 2018 Jeong and Tiwari.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 01 October 2017
                : 15 January 2018
                Page count
                Figures: 1, Tables: 1, Equations: 0, References: 71, Pages: 9, Words: 6295
                Categories
                Neuroscience
                Mini Review

                Neurosciences
                epigenetics,neurogenesis,development,neocortex,stem cells
                Neurosciences
                epigenetics, neurogenesis, development, neocortex, stem cells

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