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      Cross-Species Analysis of Single-Cell Transcriptomic Data

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          Abstract

          The ability to profile hundreds of thousands to millions of single cells using scRNA-sequencing has revolutionized the fields of cell and developmental biology, providing incredible insights into the diversity of forms and functions of cell types across many species. These technologies hold the promise of developing detailed cell type phylogenies which can describe the evolutionary and developmental relationships between cell types across species. This will require sampling of many species and taxa using single-cell transcriptomics, and methods to classify cell type homologies and diversifications. Many tools currently exist for analyzing single cell data and identifying cell types. However, cross-species comparisons are complicated by many biological and technical factors. These factors include batch effects common to deep-sequencing approaches, well known evolutionary relationships between orthologous and paralogous genes, and less well-understood evolutionary forces shaping transcriptome variation between species. In this review, I discuss recent developments in computational methods for the comparison of single-cell-omic data across species. These approaches have the potential to provide invaluable insight into how evolutionary forces act at the level of the cell and will further our understanding of the evolutionary origins of animal and cellular diversity.

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          Quantitative single-cell RNA-seq with unique molecular identifiers.

          Single-cell RNA sequencing (RNA-seq) is a powerful tool to reveal cellular heterogeneity, discover new cell types and characterize tumor microevolution. However, losses in cDNA synthesis and bias in cDNA amplification lead to severe quantitative errors. We show that molecular labels--random sequences that label individual molecules--can nearly eliminate amplification noise, and that microfluidic sample preparation and optimized reagents produce a fivefold improvement in mRNA capture efficiency.
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            Molecular identity of human outer radial glia during cortical development.

            Radial glia, the neural stem cells of the neocortex, are located in two niches: the ventricular zone and outer subventricular zone. Although outer subventricular zone radial glia may generate the majority of human cortical neurons, their molecular features remain elusive. By analyzing gene expression across single cells, we find that outer radial glia preferentially express genes related to extracellular matrix formation, migration, and stemness, including TNC, PTPRZ1, FAM107A, HOPX, and LIFR. Using dynamic imaging, immunostaining, and clonal analysis, we relate these molecular features to distinctive behaviors of outer radial glia, demonstrate the necessity of STAT3 signaling for their cell cycle progression, and establish their extensive proliferative potential. These results suggest that outer radial glia directly support the subventricular niche through local production of growth factors, potentiation of growth factor signals by extracellular matrix proteins, and activation of self-renewal pathways, thereby enabling the developmental and evolutionary expansion of the human neocortex.
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              Molecular Diversity of Midbrain Development in Mouse, Human, and Stem Cells

              Summary Understanding human embryonic ventral midbrain is of major interest for Parkinson’s disease. However, the cell types, their gene expression dynamics, and their relationship to commonly used rodent models remain to be defined. We performed single-cell RNA sequencing to examine ventral midbrain development in human and mouse. We found 25 molecularly defined human cell types, including five subtypes of radial glia-like cells and four progenitors. In the mouse, two mature fetal dopaminergic neuron subtypes diversified into five adult classes during postnatal development. Cell types and gene expression were generally conserved across species, but with clear differences in cell proliferation, developmental timing, and dopaminergic neuron development. Additionally, we developed a method to quantitatively assess the fidelity of dopaminergic neurons derived from human pluripotent stem cells, at a single-cell level. Thus, our study provides insight into the molecular programs controlling human midbrain development and provides a foundation for the development of cell replacement therapies.
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                Author and article information

                Contributors
                Journal
                Front Cell Dev Biol
                Front Cell Dev Biol
                Front. Cell Dev. Biol.
                Frontiers in Cell and Developmental Biology
                Frontiers Media S.A.
                2296-634X
                02 September 2019
                2019
                : 7
                : 175
                Affiliations
                [1] 1Biozentrum, University of Basel , Basel, Switzerland
                [2] 2Department of Molecular and Cellular Biology, Harvard University , Cambridge, MA, United States
                Author notes

                Edited by: Andreas Hejnol, University of Bergen, Norway

                Reviewed by: Jordi Solana, Oxford Brookes University, United Kingdom; Eve Gazave, UMR7592 Institut Jacques Monod (IJM), France

                *Correspondence: Maxwell E. R. Shafer, max.shafer@ 123456gmail.com ; maxwell.shafer@ 123456unibas.ch

                This article was submitted to Evolutionary Developmental Biology, a section of the journal Frontiers in Cell and Developmental Biology

                Article
                10.3389/fcell.2019.00175
                6743501
                30733944
                3a8117ab-dad2-4930-bfaf-3da1ddde7e4d
                Copyright © 2019 Shafer.

                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(s) 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
                : 31 March 2019
                : 12 August 2019
                Page count
                Figures: 2, Tables: 0, Equations: 0, References: 76, Pages: 9, Words: 0
                Funding
                Funded by: Canadian Institutes of Health Research 10.13039/501100000024
                Categories
                Cell and Developmental Biology
                Perspective

                evolutionary cell biology,single-cell rna sequencing,transcriptome evolution,species comparisons,cell types

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