The diverse subtypes of excitatory neurons that populate the neocortex are born from apical progenitors located in the ventricular zone. During corticogenesis, apical progenitors sequentially generate deep-layer neurons followed by superficial-layer neurons directly or via the generation of intermediate progenitors. Whether neurogenic fate progression necessarily implies fate restriction in single progenitor types is unknown. Here we specifically isolated apical progenitors and intermediate progenitors, and fate-mapped their respective neuronal progeny following heterochronic transplantation into younger embryos. We find that apical progenitors are temporally plastic and can re-enter past molecular, electrophysiological and neurogenic states when exposed to an earlier-stage environment by sensing dynamic changes in extracellular Wnt. By contrast, intermediate progenitors are committed progenitors that lack such retrograde fate plasticity. These findings identify a diversity in the temporal plasticity of neocortical progenitors, revealing that some subtypes of cells can be untethered from their normal temporal progression to re-enter past developmental states.
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The datasets generated in this study are available in the GEO repository under accession number GSE122644.
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We thank members of the Jabaudon laboratory and E. Azim for comments on the manuscript. We thank O. Raineteau for providing plasmids. We thank A. S. Lopes, A. Benoit, the FACS facility, the Genomics Platform and the Bioimaging Facility of the University of Geneva for technical assistance. Work in the Jabaudon laboratory is supported by the Swiss National Science Foundation, the Fondation des HUG, and the Carigest Foundation. P.O. is supported by a fellowship from iGE3.
: The authors declare no competing interests.
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Peer review information Nature thanks André Goffinet and the other anonymous reviewer(s) for their contribution to the peer review of this work.
Extended data figures and tables
a, FACS of FlashTag+ cells 1 h after labelling. b, Donor APs integrate the host VZ at discrete sites. Only a few APs (median = 2) are present at each site, such that daughter neurons found at single integration sites at P7 are probably born from a small number of initial APs (oligoclonal analysis, Figs. 1g, 4d, Extended Data Fig. 8b). c, Left, photomicrograph of a transplanted AP showing a radial glia morphology (maximum projection). Right, juxtaventricular mitosis in a transplanted AP. Experiments in a–c were repeated at least three times with similar results. d, The progeny of transplanted APs progressively migrate towards the cortical plate. The time course of this migration is similar to that of the host cells, as assessed by comparison with the migration of FlashTag-labelled endogenous cells. Box plots show median and interquartile range. n = 46 cells (RFP 6 h), 21 cells (RFP 12 h), 30 cells (RFP 24 h), 30 cells (RFP 48 h), 31 cells (RFP 72 h), 36 cells (FlashTag 6 h), 25 cells (FlashTag 12 h), 33 cells (FlashTag 24 h), 39 cells (FlashTag 48 h), 31 cells (FlashTag 72 h).
Extended Data Fig. 2 Analysis of single integration sites in AP15→15 and AP12→12—the transplantation procedure does not affect the neurogenic competence of APs.
a, Isochronically transplanted E15.5 APs (AP15→15) essentially generate SL neurons. Photomicrographs: within each donor litter, illustrations are clustered by integration site. Where applicable, a vertical black line delineates distinct host pups within a given litter. Experiments were repeated eight times with similar results. b, Isochronically transplanted E12.5 APs (AP12→12) generate DL and SL neurons. Photomicrographs: within each donor litter, illustrations are clustered by integration site. Where applicable, a vertical black line delineates distinct host pups within a given litter. Experiments were repeated six times with similar results. c, The laminar distribution of daughter neurons in the AP15→15 condition is replicated by in utero electroporation of a piggyBac-transposon construct at E15.5, in the absence of transplantation. d, The laminar distribution of daughter neurons in the AP12→12 condition is replicated by in utero electroporation of a piggyBac-transposon construct at E12.5. c, d, Box plots show median and interquartile range. In bar graphs, values are shown as mean ± s.e.m.; n = 5 pups (EporPB15), 7 experimental litters (AP15→15), 3 pups (EporPB12), 6 experimental litters (AP12→12). Two-way ANOVA with post hoc Tukey test. Epor, in utero electroporation. AP15→15 and AP12→12 distribution plots are reproduced from Fig. 1.
Extended Data Fig. 3 Analysis of single integration sites in AP15→12 and lack of temporal respecification in gliogenic AP17→12.
a, E15.5 APs transplanted into an E12.5 host (AP15→12) generate DL and SL neurons. Photomicrographs: within each donor litter, illustrations are clustered by integration site. Where applicable, a vertical black line delineates distinct host pups within a given litter. Experiments were repeated six times with similar results. b, Laminar distribution of daughter neurons across conditions. AP15→15 and AP12→12 distribution plots are reproduced from Extended Data Fig. 2 to enable direct comparison across conditions. Values are shown as mean ± s.e.m.; n = 7 experimental litters (AP15→15), 6 experimental litters (AP12→12), 8 experimental litters (AP15→12). Two-way ANOVA with post hoc Tukey test. c, E17.5 APs transplanted into an E12.5 host (AP17→12) still almost exclusively generate glial cells. Arrowheads show examples of GFAP+ cells. Values are shown as mean ± s.e.m.; n = 3 experimental litters per group. Two-tailed t-test. ***P < 0.001, ****P < 0.0001.
Extended Data Fig. 4 Molecular markers of AP15→12 daughter neurons and approach used to identify transplanted nascent neurons.
a, Reduced expression of the SL marker CUX1 in DL AP15→12 daughter neurons. b, Increased expression of the DL marker CTIP2 in DL AP15→12 daughter neurons. c, Increased expression of the DL marker TBR1 in DL AP15→12 daughter neurons. Photomicrographs showing pattern of expression of CUX1, CTIP2 and TBR1 are reproduced from Fig. 2. AP15→12 distribution plots in a–c have been reproduced from Fig. 2 for direct comparison. d, Left, schematic of the chronic EdU-labelling approach used to distinguish between nascent donor neurons and neurons born in the host. Centre, photomicrograph showing examples of an EdU+ and an EdU− donor neuron. Right, quantification of the fraction of EdU− labelled neurons at P7 (that is, transplanted cells that never underwent division in the host). Experiments were repeated three times with similar results. e, Heterochronically transplanted E15.5 nascent neurons (N15→12) migrate to the superficial layers, as they would have done in their original host. Values are shown as mean ± s.e.m.; n = 3 experimental litters (N15→12), 8 experimental litters (AP15→12). AP15→12 data in e have been reproduced from Fig. 1f for direct comparison. M, mitosis; S, S phase.
Extended Data Fig. 5 Repression of AP15-type transcriptional programs and re-induction of AP12-type transcriptional programs in AP15→12.
a, SVM classification of AP15 and AP15→12. Box plots indicate median and interquartile range. n = 26 cells (AP15), 19 cells (AP15→12). b, Expression of the AP15 transcripts used in the model. c, Expression of the AP15→12 transcripts used in the model. In b, c, n = 20 cells (AP12), 26 cells (AP15), 19 cells (AP15→12). d, Examples of hybridizations from the Allen Developing Mouse Brain Atlas (© 2008 Allen Institute for Brain Science. Allen Developing Mouse Brain Atlas. Available from: http://developingmouse.brain-map.org/) corresponding to genes shown in Fig. 3d. e, The expression of potassium channels is repressed in AP15→12. n = 20 cells (AP12), 26 cells (AP15), 19 cells (AP15→12).
a, Parameters used to define AP12→15 identity. Letters refer to panels in this figure. b, The resting membrane potential of AP12→15 remains at AP12 values. Box plots indicate median and interquartile range; n = 27 cells (AP15), 21 cells (AP12), 12 cells (AP12→15). Kruskal–Wallis test with post hoc Dunn’s test. c, Neurogenic divisions in AP12→15 remain at AP12 values. n = 3 experimental litters per group. One-way ANOVA with post hoc Tukey test. d, Number of cells per integration site at P7. Each point represents one oligoclone. The oligoclone size of AP12→15 remains at AP12→12 values. n = 37 oligoclones (AP15→15), 54 oligoclones (AP12→12), 78 oligoclones (AP12→15). Kruskal–Wallis test with post hoc Dunn’s test. AP15, AP12, AP12→12 and AP15→15 data in b–d have been reproduced from Fig. 3 for direct comparison. e, AP12→15 still generate CTIP2+ daughter neurons. n = 3 experimental litters per group. f, AP12→15 still generate TBR1+ daughter neurons. n = 3 experimental litters (AP15→15 and AP12→12), 2 experimental litters (AP12→15). g, Mismigration of CUX1-expressing daughter neurons into DL in AP12→15. n = 3 experimental litters (AP12→12), 4 experimental litters (AP12→15). Photomicrographs showing pattern of expression of CUX1, CTIP2 and TBR2 are reproduced from Fig. 2. AP12→12 and AP15→15 data in e–g have been reproduced from Fig. 2 for direct comparison. Data in c–g are shown as mean ± s.e.m. One-way ANOVA with post hoc Tukey test (e–g). h, EdU pulse labelling at E17 and E18 shows progressive mismigration of increasingly later-born neurons in AP12→15. Box plots show mean and interquartile range. n = 19 cells (E17), 12 cells (E18). i, Summary of the mismigration phenotype in AP12→15. **P < 0.01, ***P < 0.001.
E12.5 APs transplanted into an E15.5 host (AP12→15) generate DL and SL neurons. Photomicrographs: within each donor litter, illustrations are clustered by integration site. Where applicable, a vertical black line delineates distinct host pups within a given litter. Experiments were repeated six times with similar results.
a, Schematic showing EdU-based labelling and transplantation procedure. b, E15.5 EdU+ cells transplanted into an E12.5 host (EdU15→12) still give rise to SL neurons (compare with Fig. 1d). Box plot shows median and interquartile range. Top right, modal distribution of daughter neurons in SL vs DL. Values are shown as mean ± s.e.m.; n = 3 experimental litters. Bottom right, laminar distribution of daughter neurons at single integration sites. Values refer to number of integration sites in each category. c, Heterochronically transplanted EdU-labelled progenitors (EdU15→12) essentially generate SL neurons. Photomicrographs: within each donor litter, illustrations are clustered by integration site. Experiments were repeated three times with similar results. d, Ten hours after FlashTag labelling, most cells have differentiated into IPs (that is, KI67+TBR2+ cells). Values are shown as mean ± s.e.m.; n = 3 pups. e, IP15→12 essentially give rise to SL neurons. Photomicrographs: within each donor litter, illustrations are clustered by integration site. Only EdU+ neurons (filled arrowheads) were included in this analysis. Experiments were repeated three times with similar results. f, Number of cells per integration site at P7. Each point represents one oligoclone. AP15→12 data has been reproduced from Fig. 3g for direct comparison. Values are shown as mean ± s.e.m.; n = 39 oligoclones (AP15→12), 17 oligoclones (IP15→12). g, IP15→12 daughter neurons express CUX1 but not CTIP2. Photomicrographs showing pattern of expression of CUX1 and CTIP2 are reproduced from Fig. 2. Values are shown as mean ± s.e.m.; N = 2 experimental litters (CUX1), 3 experimental litters (CTIP2). In b, vertically aligned cells belong to single integration sites. FT, FlashTag; IZ, intermediate zone.
a, Top, temporally dynamic expression of Wnt transcripts in the developing cortex (data from www.unige.ch/genebrowser.unige.ch/telagirdon)8. Values are shown as mean ± s.e.m.; n = 189 APs, 268 neurons (E12), 207 APs, 223 neurons (E13), 134 APs, 219 neurons (E14), 301 APs, 213 neurons (E15). Right, example of corresponding in situ hybridizations from the Allen Developing Mouse Brain Atlas (© 2008 Allen Institute for Brain Science. Allen Developing Mouse Brain Atlas. Available from: http://developingmouse.brain-map.org/). b, Dynamic expression of Wnt transcripts in the developing cortex (expression landscapes from www.unige.ch/genebrowser.unige.ch/telagirdon)8. The overall pattern is early–high to late–low (a). c, AP15 express Wnt receptor-related transcriptional programs (data from ref. 8). Box plots indicate median and interquartile range. Right, expression of individual receptor-related transcripts in AP12 and AP15. n = 189 APs (E12), 301 APs (E15).
This file contains Supplementary Notes 1-3, Discussion and References.
Supplementary Table 1| Source data of radial position, laminar location and oligoclone identity of all neurons in this study. This file contains the values for the normalized radial position, laminar location and oligoclone identity of all the neurons used for analysis in the following conditions: AP15→15, AP12→12, AP15→12, EdU15→12, IP15→12 and AP12→15.
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Oberst, P., Fièvre, S., Baumann, N. et al. Temporal plasticity of apical progenitors in the developing mouse neocortex. Nature 573, 370–374 (2019). https://doi.org/10.1038/s41586-019-1515-6
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