RT Journal Article
SR Electronic
T1 Decoding the molecular landscape of the developing spatial processing system and production of entorhinal stellate cell-like cells by a direct programming approach
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 738443
DO 10.1101/738443
A1 Tobias Bergmann
A1 Yong Liu
A1 Leo Mogus
A1 Julie Lee
A1 Ulrich Pfisterer
A1 Louis-Francois Handfield
A1 Andrea Asenjo-Martinez
A1 Irene Lisa-Vargas
A1 Stefan E Seemann
A1 Jimmy Tsz Hang Lee
A1 Nikolaos Patikas
A1 Birgitte Rahbek Kornum
A1 Mark Denham
A1 Poul Hyttel
A1 Menno P Witter
A1 Jan Gorodkin
A1 Tune H Pers
A1 Martin Hemberg
A1 Konstantin Khodosevich
A1 Vanessa Jane Hall
YR 2021
UL http://biorxiv.org/content/early/2021/03/31/738443.abstract
AB Classic studies investigating how and when the entorhinal cortex (component of the memory processing system of the brain) develops have been based on traditional thymidine autoradiography and histological techniques. In this study, we take advantage of modern technologies to trace at a high resolution, the cellular complexity of the developing porcine medial entorhinal cortex by using single-cell profiling. The postnatal medial entorhinal cortex comprises 4 interneuron, 3 pyramidal neuron and 2 stellate cell populations which emerge from intermediate progenitor and immature neuron populations. We discover four MGE-derived interneurons and one CGE-derived interneuron population as well as several IN progenitors. We also identify two oligodendrocyte progenitor populations and three populations of oligodendrocytes. We perform a proof-of-concept experiment demonstrating that porcine scRNA-seq data can be used to develop novel protocols for producing human entorhinal cells in-vitro. We identified six transcription factors (RUNX1A1, SOX5, FOXP1, MEF2C, TCF3, EYA2) important in neurodevelopment and differentiation from one RELN+ stellate cell population. Using a lentiviral vector approach, we reprogrammed human induced pluripotent stem cells into stellate cell-like cells which expressed RELN, SATB2, LEF1 and BCL11B. Our findings contribute to the understanding of the formation of the brain’s cognitive memory and spatial processing system and provides proof-of-concept for the production of entorhinal cells from human pluripotent stem cells in-vitro.Competing Interest StatementThe authors have declared no competing interest.AbbreviationsBMPbone morphogenetic proteinCHIRGSK-3α/β inhibitorCNScentral nervous systemDDayDMEMdulbecco’s modified eagle’s mediumEEmbryonic dayECentorhinal cortexEDTAEthylenediaminetetraacetic acidgDNAgenomic DNAGOIgene of interesthhoursiPSCinduced pluripotent stem cellsINinterneuronsIPintermediate progenitorLlayerLVlentivirusLEClateral entorhinal cortexMECmedial entorhinal cortexMOImultiplicity of infectionOPColigodendrocyte progenitor cellPpostnatal dayPBSphosphate-buffered salinePCRpolymerase chain reactionPYRpyramidal neuronqPCRquantitative real time PCRRELNReelin geneRTroom temperaturescRNA-seqSingle-cell RNA sequencingSCstellate cellSDstandard deviationSPEspeed cellSSTsomatostatinTFtranscription factorUunits of viral particlesUMIunique molecular identifierVPDvolume of produced virus