Neural Crest Development PDF

Neural Crest Development Lukas Sommer, Stem Cell Biology, Institute of Anatomy, University of Zurich  Neurulation Amnion 19 – 23 d 6 1 4...

Neural Crest Development Lukas Sommer, Stem Cell Biology, Institute of Anatomy, University of Zurich  Neurulation Amnion 19 – 23 d 6 1 4 3 3 1 Neural plate 2 Primitive streak 2 3 Primitive node 4 Neural groove 5 Somites 6 Amnion 7 Neural folds https://www.youtube.com/watch?v=xAxqdeVrhJI Generation of Neural Crest Cells during Neurulation https://www.youtube.com/watch?v=FhhWG3XzARY (0’43’’ to 2’30’’) The Neural Crest as a Model System to Study Stem Cell Biology Highly migratory cell population in the vertebrate embryo emerging at the dorsal part of the closing neural tube during neurulation Studying epithelial-to-mesenchymal transition (EMT) and migration of Neural Crest Cells  The Neural Crest Cranial Neural Crest Cardiac Neural Crest Vagal Neural Crest Trunk Neural Crest Nessy John  Neural Crest Targets Pharyngeal Apparatus Outflow Tract of Heart Pigment Cells underneath the Epidermis Enteric Ganglia (ENS) Sympathetic Ganglia (PNS) Dorsal Root Ganglia (PNS) Nessy John The Neural Crest as a Model System to Study Stem Cell Biology Neural crest cells generate most of the peripheral nervous system, pigment cells in the skin, smooth muscle in the outflow tract of the heart, craniofacial bone and cartilage, etc  Mapping cell derivatives with quail chick chimeras Le Douarin Quail Chick NCC derivatives Identically staged chick in ovo Garcia Castro  In vivo cell fate mapping using Cre-recombinase-mediated recombination in mice http://www.ruf.rice.edu/~rur/issue1_files/norman.html https://www.jax.org/news-and-insights/jax-blog/2011/october/cre- lox-breeding-for-dummies-part-ii In vivo cell fate mapping using Cre-recombinase-mediated recombination in mice (driven from ubiquitous promoter) https://www.jax.org/news-and-insights/jax-blog/2011/october/cre- lox-breeding-for-dummies-part-ii -> Expression of Cre recombinase in the area of Wnt1 expression In vivo Fate Mapping of Neural Crest Cells : e.g. to the outflow tract of the heart Jiang et al., Development 2000  Neural Crest Cells and their Derivatives dorsal Coelomhöhle ventral Neural : Non-Neural : Glia (Schwann cells; satellite glia Smooth muscle (outflow tract of the heart) Sensory nervous system (dorsal root ganglia) Adrenal medulla (Chromaffin cells) Autonomic nervous system Melanocytes (Sympathicus, Parasympathicus) Craniofacial bones and cartilage Enteric nervous system Odontoblasts (producing dentin in teeth)  Cranial neural crest cells and their derivatives Martik, M.L., Bronner, M.E. Nat Rev Neurosci (2021) https://doi.org/10.1038/s41583-021-00503-2  Early derivatives of neural crest cells: Schwann cell precursors (SCP) associated with peripheral axons SCP https://doi.org/10.1002/wdev.398 Fate mapping with lines expressing Cre at a somewhat later time point than Wnt1-Cre reveal «Schwann cell precursors» to have a potential probably as broad as that of migratory neural crest cells Fate mapping with lines expressing Cre at a somewhat later time point than Wnt1-Cre reveal «Schwann cell precursors» to have a potential probably as broad as that of migratory neural crest cells  Fate vs. Potential In vivo NCCs differentiate according to their axial position NCC derivatives Cranial Fates of neural crest cells influenced by new environmental signals / axial positions… Garcia Castro  how to distinguish between these mechanisms ?  Clonal analysis in vitro : The Neural Crest Culture System E9 Mouse Embryo NC explant NC explant, neural tube removed  The Neural Crest Cell Culture System : “Neural Crest Stem Cells (NCSCs)” Stemple and Anderson, Cell 1992: Prospective identification based on expression of a surface protein (allowing live cell staining) At least an “in vitro stem cell”, similar to ES cells and others...  Neural Crest Cells in vivo: homogeneous population of multipotent cells or heterogeneous population of fate-restricted cells ? Multipotent NC Restricted heterogeneous NC  Clonal analysis of premigratory vs migratory Neural Crest Cells in vivo Low titer retroviral infection Single cell dye labelling in chicken embryos -> Premigratory and migratory multipotent neural crest cells in chicken and mice (Marianne Bronner; Josh Sanes, early `80ties) -> With an adapted protocol, however : premigratory neural crest cells are lineage-restricted in chicken (Chaya Kalcheim, 2010) -> Single cell genetic cell fate mapping using „confetti mice“ confirms multipotency of neural crest cells in mice  Tracing premigratory (Wnt1-CreERT) Wnt1-CreERT Wnt1-CreERT R26R Tracing neural crest cells using confetti mice Wnt1CreERT2 R26R With tamoxifen E9 Hsp 90 Cre ER E9.5 E12.5 Wnt1-CreERT C Active D Sox10CreERT2 R26R E12.5 E9.5 E CAGG promoter R26R pGK Neor GFP RFP YFP CFP G H  Addressing Multipotency in vivo: Tracing of Single Premigratory NC Cells using a Multicolor Cre Reporter “Confetti” homozygous Wnt1-CreERT R26R-Confetti  Quantitative analysis of clones spread over a defined segment of the trunk  Quantitative analysis of rare color clones at low clonal density In mice, the vast majority of neural crest cells appear to be multipotent at the stage analyzed, with very few clones contributing to single derivatives. Intriguingly, multipotency appears to be maintained in migratory neural crest cells: Evidence for stem cells in vivo Baggiolini et al, Cell Stem Cell 2015  The power of single cell RNA sequencing (scRNAseq): Addressing cellular identities and developmental trajectories of neural crest cells by scRNAseq Embryo, in which neural crest cells have been genetically traced (e.g. Wnt1-Cre :: Various methods for scRNAseq; see e.g. tdTomato) https://www.10xgenomics.com/products/single-cell-gene-expression Bioinformatics tools to define “cell types” (clusters of gene co- expression patterns) and how one cell type gives rise to another one (“developmental trajectories”)  The power of single cell RNA sequencing: Addressing cellular identities and developmental trajectories of neural crest cells by scRNAseq Despite broad developmental potential in many cells (see Confetti study), there is a quite extensive molecular heterogeneity at the single cell level; Neural crest cells differentiate through sequential lineage-restriction events, involving co-expression and competition of genes driving alternative fates. Soldatov et al., (Adameyko lab), Science 2019 Neural Crest-Derived Cells with Stem Cell Features Also Persist in Adult Structures In vivo fate mapping and prospective identification and isolation Shakhova and Sommer, StemBook 2010 Neural crest-derived cells in adult skin are multipotent  Emergence of Neural Crest Stem Cell-like cells in adult skin - In analogy to other adult stem cells, physiological role in homeostasis and regeneration (wound healing) - Role in initiation of tumors derived from neural crest structures (e.g. neuroblastoma; melanoma)  Tumors hijacking embryonic programs: Neural Crest Stem Cells States in melanoma Diener and Sommer, Stem Cells Translational Medicine, 2021