Article | Published:

Layer V cortical neurons require microglial support for survival during postnatal development

Nature Neuroscience volume 16, pages 543551 (2013) | Download Citation

Abstract

Neurons require trophic support during neural circuit formation; however, how the cellular milieu contributes to neuronal survival remains unclear. We found that layer V cortical neurons require support from microglia for survival during postnatal development. Specifically, we found that microglia accumulated close to the subcerebral and callosal projection axons in the postnatal brain. Inactivation of microglia by minocycline treatment or transient ablation of microglia in CD11b-DTR transgenic mice led to increased apoptosis, specifically in layer V subcerebral and callosal projection neurons. CX3CR1 in microglia was required for the survival of layer V neurons. Microglia consistently promoted the survival of cortical neurons in vitro. In addition, we identified microglia-derived IGF1 as a trophic factor that maintained neuronal survival. Our results highlight a neuron-glia interaction that is indispensable for network formation during a specific period in the developing brain.

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References

  1. 1.

    & Programmed Cell Death and Neurotrophic Factors (Academic Press, 2003).

  2. 2.

    & Neurotrophins: roles in neuronal development and function. Annu. Rev. Neurosci. 24, 677–736 (2001).

  3. 3.

    , , & Specific neurotrophic factors support the survival of cortical projection neurons at distinct stages of development. J. Neurosci. 21, 8863–8872 (2001).

  4. 4.

    & Connectional distinction between callosal and subcortically projecting cortical neurons is determined prior to axon extension. Dev. Biol. 160, 1–14 (1993).

  5. 5.

    et al. Satb2 regulates callosal projection neuron identity in the developing cerebral cortex. Neuron 57, 364–377 (2008).

  6. 6.

    et al. Neuronal subtype–specific genes that control corticospinal motor neuron development in vivo. Neuron 45, 207–221 (2005).

  7. 7.

    & IGF-I specifically enhances axon outgrowth of corticospinal motor neurons. Nat. Neurosci. 9, 1371–1381 (2006).

  8. 8.

    et al. A novel purification method for CNS projection neurons leads to the identification of brain vascular cells as a source of trophic support for corticospinal motor neurons. J. Neurosci. 28, 8294–8305 (2008).

  9. 9.

    , & Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo. Science 308, 1314–1318 (2005).

  10. 10.

    , , , & Selective ablation of proliferating microglial cells exacerbates ischemic injury in the brain. J. Neurosci. 27, 2596–2605 (2007).

  11. 11.

    , & Debris clearance by microglia: an essential link between degeneration and regeneration. Brain 132, 288–295 (2009).

  12. 12.

    , & Engulfment of axon debris by microglia requires p38 MAPK activity. J. Biol. Chem. 284, 21626–21636 (2009).

  13. 13.

    , , , & Resting microglia directly monitor the functional state of synapses in vivo and determine the fate of ischemic terminals. J. Neurosci. 29, 3974–3980 (2009).

  14. 14.

    , , & Activated microglia inhibit axonal growth through RGMa. PLoS ONE 6, e25234 (2011).

  15. 15.

    , & The origin and cell lineage of microglia: new concepts. Brain Res. Rev. 53, 344–354 (2007).

  16. 16.

    et al. Fate mapping analysis reveals that adult microglia derive from primitive macrophages. Science 330, 841–845 (2010).

  17. 17.

    et al. Hematopoietic origin of pathological grooming in Hoxb8 mutant mice. Cell 141, 775–785 (2010).

  18. 18.

    , & Microglia derive from progenitors, originating from the yolk sac, and which proliferate in the brain. Brain Res. Dev. Brain Res. 117, 145–152 (1999).

  19. 19.

    , & Differential immunochemical markers reveal the normal distribution of brain macrophages and microglia in the developing rat brain. J. Comp. Neurol. 314, 125–135 (1991).

  20. 20.

    , , & Microglia: its development and role as a neuropathology sensor. Prog. Brain Res. 132, 61–79 (2001).

  21. 21.

    et al. Activation and deactivation of periventricular white matter phagocytes during postnatal mouse development. Glia 58, 11–28 (2010).

  22. 22.

    Microglia and macrophages in the developing CNS. Neurotoxicology 22, 619–624 (2001).

  23. 23.

    et al. Lentivirus-based genetic manipulations of cortical neurons and their optical and electrophysiological monitoring in vivo. Proc. Natl. Acad. Sci. USA 101, 18206–18211 (2004).

  24. 24.

    , , , & Tetracyclines inhibit microglial activation and are neuroprotective in global brain ischemia. Proc. Natl. Acad. Sci. USA 95, 15769–15774 (1998).

  25. 25.

    , , , & Minocycline, a tetracycline derivative, is neuroprotective against excitotoxicity by inhibiting activation and proliferation of microglia. J. Neurosci. 21, 2580–2588 (2001).

  26. 26.

    et al. Selective depletion of macrophages reveals distinct, opposing roles during liver injury and repair. J. Clin. Invest. 115, 56–65 (2005).

  27. 27.

    et al. Diphtheria toxin–induced autophagic cardiomyocyte death plays a pathogenic role in mouse model of heart failure. J. Biol. Chem. 279, 41095–41103 (2004).

  28. 28.

    , , & Persistence of early-generated neurons in the rodent subplate: assessment of cell death in neocortex during the early postnatal period. J. Neurosci. 15, 5014–5024 (1995).

  29. 29.

    Chemokines and chemokine receptors: standing at the crossroads of immunobiology and neurobiology. Immunity 31, 711–721 (2009).

  30. 30.

    et al. Control of microglial neurotoxicity by the fractalkine receptor. Nat. Neurosci. 9, 917–924 (2006).

  31. 31.

    et al. Analysis of fractalkine receptor CX(3)CR1 function by targeted deletion and green fluorescent protein reporter gene insertion. Mol. Cell Biol. 20, 4106–4114 (2000).

  32. 32.

    Serum-free B27/neurobasal medium supports differentiated growth of neurons from the striatum, substantia nigra, septum, cerebral cortex, cerebellum, and dentate gyrus. J. Neurosci. Res. 42, 674–683 (1995).

  33. 33.

    , , , & Inhibition of cellular proliferation by peptide analogues of insulin-like growth factor 1. Cancer Res. 52, 6447–6451 (1992).

  34. 34.

    & Cellular actions of the insulin-like growth factor binding proteins. Endocr. Rev. 23, 824–854 (2002).

  35. 35.

    & Microglia-derived nerve growth factor causes cell death in the developing retina. Neuron 20, 35–41 (1998).

  36. 36.

    et al. Microglia promote the death of developing Purkinje cells. Neuron 41, 535–547 (2004).

  37. 37.

    , , , & Endogenous microglia regulate development of embryonic cortical precursor cells. J. Neurosci. Res. 89, 286–298 (2011).

  38. 38.

    et al. Synaptic pruning by microglia is necessary for normal brain development. Science 333, 1456–1458 (2011).

  39. 39.

    et al. Postnatal growth of corticospinal axons in the spinal cord of developing mice. Brain Res. Dev. Brain Res. 112, 189–204 (1999).

  40. 40.

    & Molecular mechanisms of axon guidance in the developing corticospinal tract. Prog. Neurobiol. 85, 214–235 (2008).

  41. 41.

    et al. Activity-dependent development of callosal projections in the somatosensory cortex. J. Neurosci. 27, 11334–11342 (2007).

  42. 42.

    & Cortical axons branch to multiple subcortical targets by interstitial axon budding: implications for target recognition and “waiting periods”. Neuron 1, 901–910 (1988).

  43. 43.

    , , , & Igf1 gene disruption results in reduced brain size, CNS hypomyelination, and loss of hippocampal granule and striatal parvalbumin-containing neurons. Neuron 14, 717–730 (1995).

  44. 44.

    , , , & Insulin-like growth factor binding protein-1–6 expression in activated microglia. Neuroreport 15, 1033–1037 (2004).

  45. 45.

    & Expression pattern and neurotrophic role of the c-fms proto-oncogene M-CSF receptor in rodent Purkinje cells. J. Neurosci. 18, 10481–10492 (1998).

  46. 46.

    et al. GDNF-induced osteopontin from Muller glial cells promotes photoreceptor survival in the Pde6brd1 mouse model of retinal degeneration. Glia 59, 821–832 (2011).

  47. 47.

    et al. The P2Y12 receptor regulates microglial activation by extracellular nucleotides. Nat. Neurosci. 9, 1512–1519 (2006).

  48. 48.

    & Axon retraction and degeneration in development and disease. Annu. Rev. Neurosci. 28, 127–156 (2005).

  49. 49.

    & Strategies for regenerating injured axons after spinal cord injury - insights from brain development. Biologics 2, 253–264 (2008).

  50. 50.

    , , & Intraspinal rewiring of the corticospinal tract requires target-derived BDNF and compensates lost function after brain injury. Brain 135, 1253–1267 (2012).

  51. 51.

    et al. Conditional macrophage ablation demonstrates that resident macrophages initiate acute peritoneal inflammation. J. Immunol. 174, 2336–2342 (2005).

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Acknowledgements

We thank H. Ozawa for experimental technical support, M.K. Yamada (University of Tokyo), P. Osten (Cold Spring Harbor Laboratory) and H. Miyoshi (RIKEN) for their kind gift of the lentivirus plasmid and helpful suggestions, and Y. Yoshida (Cincinnati Children's Hospital Medical Center) for critical comments on the manuscript. This work was supported by a grant for Core Research for Evolutional Science and Technology (CREST) from the Japan Science and Technology Agency (JST) to T.Y. and a Grant-in-Aid for Young Scientists (B) from Japan Society for the Promotion of Science to M.U.

Author information

Author notes

    • Masaki Ueno
    •  & Yuki Fujita

    These authors contributed equally to this work.

Affiliations

  1. Department of Molecular Neuroscience, Graduate School of Medicine, Osaka University, Osaka, Japan.

    • Masaki Ueno
    • , Yuki Fujita
    • , Tatsuhide Tanaka
    • , Yuka Nakamura
    •  & Toshihide Yamashita
  2. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.

    • Masaki Ueno
    • , Yuki Fujita
    • , Yuka Nakamura
    •  & Toshihide Yamashita
  3. Department of Functional Anatomy and Neuroscience, Asahikawa Medical University, Hokkaido, Japan.

    • Tatsuhide Tanaka
  4. Laboratory of Cellular Dynamics, Immunology Frontier Research Center, Osaka University, Osaka, Japan.

    • Junichi Kikuta
    •  & Masaru Ishii

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Contributions

M.U. and T.Y. conceived the project, designed the experiments and wrote the paper. M.U. and Y.F. performed the experiments and analyzed the data. T.T. contributed to the culture experiments, and Y.N. contributed to the culture experiments and in situ hybridization. J.K. and M.I. contributed to the experiments using Cx3cr1-deficient mice. T.Y. coordinated and directed the project.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Masaki Ueno or Toshihide Yamashita.

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DOI

https://doi.org/10.1038/nn.3358

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