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Transplanted embryonic neurons integrate into adult neocortical circuits

Nature volume 539, pages 248253 (10 November 2016) | Download Citation

  • An Erratum to this article was published on 07 December 2016

This article has been updated


The ability of the adult mammalian brain to compensate for neuronal loss caused by injury or disease is very limited. Transplantation aims to replace lost neurons, but the extent to which new neurons can integrate into existing circuits is unknown. Here, using chronic in vivo two-photon imaging, we show that embryonic neurons transplanted into the visual cortex of adult mice mature into bona fide pyramidal cells with selective pruning of basal dendrites, achieving adult-like densities of dendritic spines and axonal boutons within 4–8 weeks. Monosynaptic tracing experiments reveal that grafted neurons receive area-specific, afferent inputs matching those of pyramidal neurons in the normal visual cortex, including topographically organized geniculo-cortical connections. Furthermore, stimulus-selective responses refine over the course of many weeks and finally become indistinguishable from those of host neurons. Thus, grafted neurons can integrate with great specificity into neocortical circuits that normally never incorporate new neurons in the adult brain.

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Change history

  • 09 November 2016

    Minor changes were made to Figs 4, 5 and Extended Data Fig. 8.

  • 17 November 2016

    The resolution of Fig. 4 was increased.


  1. 1.

    , & Stem cell therapy for human neurodegenerative disorders-how to make it work. Nat. Med. 10 (suppl.), S42–S50 (2004)

  2. 2.

    & Reconstruction of brain circuitry by neural transplants generated from pluripotent stem cells. Neurobiol. Dis. 79, 28–40 (2015)

  3. 3.

    , , & Fetal dopaminergic transplantation trials and the future of neural grafting in Parkinson’s disease. Lancet Neurol. 12, 84–91 (2013)

  4. 4.

    et al. Monosynaptic tracing using modified rabies virus reveals early and extensive circuit integration of human embryonic stem cell-derived neurons. Stem Cell Reports 4, 975–983 (2015)

  5. 5.

    et al. Transplanted neuroblasts differentiate appropriately into projection neurons with correct neurotransmitter and receptor phenotype in neocortex undergoing targeted projection neuron degeneration. J. Neurosci. 20, 7404–7416 (2000)

  6. 6.

    et al. hPSC-derived maturing GABAergic interneurons ameliorate seizures and abnormal behavior in epileptic mice. Cell Stem Cell 15, 559–573 (2014)

  7. 7.

    et al. Area-specific reestablishment of damaged circuits in the adult cerebral cortex by cortical neurons derived from mouse embryonic stem cells. Neuron 85, 982–997 (2015)

  8. 8.

    & Highly selective receptive fields in mouse visual cortex. J. Neurosci. 28, 7520–7536 (2008)

  9. 9.

    & Noninvasively induced degeneration of neocortical pyramidal neurons in vivo: selective targeting by laser activation of retrogradely transported photolytic chromophore. Exp. Neurol. 121, 153–159 (1993)

  10. 10.

    Maturation of rat visual cortex. I. A quantitative study of Golgi-impregnated pyramidal neurons. J. Neurocytol. 10, 859–878 (1981)

  11. 11.

    et al. Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes. Nature 425, 968–973 (2003)

  12. 12.

    et al. Neural stem cells engrafted in the adult brain fuse with endogenous neurons. Stem Cells Dev. 22, 538–547 (2013)

  13. 13.

    et al. Cortical excitatory neurons and glia, but not GABAergic neurons, are produced in the Emx1-expressing lineage. J. Neurosci. 22, 6309–6314 (2002)

  14. 14.

    et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 13, 133–140 (2010)

  15. 15.

    et al. Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window. Nat. Protocols 4, 1128–1144 (2009)

  16. 16.

    , , & Experience leaves a lasting structural trace in cortical circuits. Nature 457, 313–317 (2009)

  17. 17.

    et al. Primary visual cortex shows laminar-specific and balanced circuit organization of excitatory and inhibitory synaptic connectivity. J. Physiol. (Lond.) 594, 1891–1910 (2016)

  18. 18.

    et al. Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons. Neuron 53, 639–647 (2007)

  19. 19.

    et al. A mesoscale connectome of the mouse brain. Nature 508, 207–214 (2014)

  20. 20.

    et al. Neural networks of the mouse neocortex. Cell 156, 1096–1111 (2014)

  21. 21.

    , , & Topographic organization of rat locus coeruleus and dorsal raphe nuclei: distribution of cells projecting to visual system structures. J. Comp. Neurol. 336, 345–361 (1993)

  22. 22.

    & Efferent connections of the caudate nucleus, including cortical projections of the striatum and other basal ganglia: an autoradiographic and horseradish peroxidase investigation in the cat. J. Comp. Neurol. 226, 28–49 (1984)

  23. 23.

    , & Ephrin-As and patterned retinal activity act together in the development of topographic maps in the primary visual system. J. Neurosci. 26, 12873–12884 (2006)

  24. 24.

    , & Mapping retinotopic structure in mouse visual cortex with optical imaging. J. Neurosci. 22, 6549–6559 (2002)

  25. 25.

    et al. Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature 499, 295–300 (2013)

  26. 26.

    et al. Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes. Nat. Methods 11, 175–182 (2014)

  27. 27.

    , & Emergence of feature-specific connectivity in cortical microcircuits in the absence of visual experience. J. Neurosci. 34, 9812–9816 (2014)

  28. 28.

    , , & Functional specialization of seven mouse visual cortical areas. Neuron 72, 1040–1054 (2011)

  29. 29.

    , & Cell biology in neuroscience: Cellular and molecular mechanisms underlying axon formation, growth, and branching. J. Cell Biol. 202, 837–848 (2013)

  30. 30.

    et al. Developmental sculpting of intracortical circuits by MHC class I H2-Db and H2-Kb. Cereb. Cortex 26, 1453–1463 (2014)

  31. 31.

    , & Delayed stabilization of dendritic spines in fragile X mice. J. Neurosci. 30, 7793–7803 (2010)

  32. 32.

    , , , & Late-stage immature neocortical neurons reconstruct interhemispheric connections and form synaptic contacts with increased efficiency in adult mouse cortex undergoing targeted neurodegeneration. J. Neurosci. 22, 4045–4056 (2002)

  33. 33.

    et al. Anatomical and functional reconstruction of the nigrostriatal pathway by intranigral transplants. Neurobiol. Dis. 35, 477–488 (2009)

  34. 34.

    , & Afferents to visually responsive grafts of embryonic occipital neocortex tissue implanted into V1 (Oc1) cortical area of adult rats. Restor. Neurol. Neurosci. 12, 13–25 (1998)

  35. 35.

    & Thalamocortical specificity and the synthesis of sensory cortical receptive fields. J. Neurophysiol. 94, 26–32 (2005)

  36. 36.

    , & Altered visual experience induces instructive changes of orientation preference in mouse visual cortex. J. Neurosci. 31, 13911–13920 (2011)

  37. 37.

    et al. Massive restructuring of neuronal circuits during functional reorganization of adult visual cortex. Nat. Neurosci. 11, 1162–1167 (2008)

  38. 38.

    et al. Development of direction selectivity in mouse cortical neurons. Neuron 71, 425–432 (2011)

  39. 39.

    & Layer-specific refinement of visual cortex function after eye opening in the awake mouse. J. Neurosci. 35, 3370–3383 (2015)

  40. 40.

    et al. Preferential electrical coupling regulates neocortical lineage-dependent microcircuit assembly. Nature 486, 113–117 (2012)

  41. 41.

    et al. Clonally related visual cortical neurons show similar stimulus feature selectivity. Nature 486, 118–121 (2012)

  42. 42.

    , & Neural crest cells retain multipotential characteristics in the developing valves and label the cardiac conduction system. Circ. Res. 98, 1547–1554 (2006)

  43. 43.

    et al.Adult generation of glutamatergic olfactory bulb interneurons. Nat. Neurosci. 12, 1524–1533 (2009)

  44. 44.

    In vivo electroporation in the embryonic mouse central nervous system. Nat. Protocols 1, 1552–1558 (2006)

  45. 45.

    , & Robust quantification of orientation selectivity and direction selectivity. Front. Neural Circuits 8, 92 (2014)

  46. 46.

    et al. Genome-wide atlas of gene expression in the adult mouse brain. Nature 445, 168–176 (2007)

  47. 47.

    & Free-D: an integrated environment for three-dimensional reconstruction from serial sections. J. Neurosci. Methods 145, 233–244 (2005)

  48. 48.

    , , & Putting a finishing touch on GECIs. Front. Mol. Neurosci. 7, 88 (2014)

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We thank D. Franzen, G. Jäger, T. Simon, V. Staiger, H. Tultschin and F. Voss for technical support, and A. Lepier for viral vector expertise. M. Sperling, P. Goltstein and A. Grade helped with hardware and software. This work was supported by the German Research Foundation (SFB 870 ‘Neuronal Circuits’: M.G., L.D., K.-K.C., T.B. and M.H.; SPP 1757: M.G. and L.D.), the Advanced ERC grant ChroNeuroRepair (M.G.), the Helmholtz Alliance Icemed (M.G.), the Boehringer Ingelheim Fonds (S.F.), and the Max Planck Society (S.F., T.B. and M.H.).

Author information

Author notes

    • Susanne Falkner
    •  & Sofia Grade

    These authors contributed equally to this work.

    • Magdalena Götz
    •  & Mark Hübener

    These authors jointly supervised this work.


  1. Max Planck Institute of Neurobiology, D-82152 Martinsried, Germany

    • Susanne Falkner
    • , Tobias Bonhoeffer
    •  & Mark Hübener
  2. Physiological Genomics, Biomedical Center, Ludwig-Maximilians University Munich, D-82152 Planegg, Germany

    • Sofia Grade
    • , Leda Dimou
    •  & Magdalena Götz
  3. Institute of Stem Cell Research, Helmholtz Center Munich, German Research Center for Environmental Health, D-85764 Neuherberg, Germany

    • Sofia Grade
    • , Leda Dimou
    •  & Magdalena Götz
  4. SYNERGY, Excellence Cluster of Systems Neurology, Biomedical Center, Ludwig-Maximilians University Munich, D-82152 Planegg , Germany

    • Leda Dimou
    •  & Magdalena Götz
  5. Max von Pettenkofer Institute and Gene Center, Ludwig-Maximilians University Munich, D-81377 Munich, Germany

    • Karl-Klaus Conzelmann


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The original idea for the study came from M.G. M.G, M.H. and T.B. then initiated the study and planned the experimental approach. S.F., S.G., M.G., M.H. and L.D. designed the experiments; S.F. performed in vivo imaging experiments and analysis; S.G. performed the experiments in fixed tissue including connectivity experiments and analysis. M.G. and L.D. provided the lesion model. K.-K.C. provided the rabies virus and expertise for its use for monosynaptic tracing. Finally, M.G., M.H., S.G. and S.F. wrote the paper with input from T.B., L.D. and K.-K.C.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Magdalena Götz or Mark Hübener.

Extended data

Supplementary information


  1. 1.

    Morphological development of a transplanted neuron

    Neuron 3 to 92 dpt (same as in Fig. 1), developing a L2/3 pyramidal cell-like morphology within 3 wpt. Stable overall morphology 4 to 13 wpt. In vivo two-photon z-stacks depicted as maximum projections, time series.

  2. 2.

    Formation of dendritic spines

    Dendrite 6 to 75 dpt, forming first dendritic spines at 9 dpt. Z-stack maximum projections, time series.

  3. 3.

    Formation of axonal boutons

    Axon 5 to 84 dpt (same as in Fig. 2), forming first axonal boutons at 5 dpt, a secondary branch forms at 7 dpt. Z-stack maximum projections, time series.

  4. 4.

    3D reconstruction of dLGN relay cells retrogradely traced in mice with distinct transplantation sites in V1

    Data from each mouse (n=8) is represented with a distinct color, and each sphere corresponds to one neuron (n=3-96/mouse). Animation starts with an anterior view and then rotates around several axes, demonstrating that cells form segregated clusters in specific parts of the dLGN (surface in wireframe; see Extended Data Fig. 8c).

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