Abstract
The balance between proliferation and apoptosis is critical for proper development of the nervous system. Yet, little is known about molecules that regulate apoptosis of proliferative neurons. Here we identify a soluble, secreted form of CPG15 expressed in embryonic rat brain regions undergoing rapid proliferation and apoptosis, and show that it protects cultured cortical neurons from apoptosis by preventing activation of caspase 3. Using a lentivirus-delivered small hairpin RNA, we demonstrate that endogenous CPG15 is essential for the survival of undifferentiated cortical progenitors in vitro and in vivo. We further show that CPG15 overexpression in vivo expands the progenitor pool by preventing apoptosis, resulting in an enlarged, indented cortical plate and cellular heterotopias within the ventricular zone, similar to the phenotypes of mutant mice with supernumerary forebrain progenitors. CPG15 expressed during mammalian forebrain morphogenesis may help balance neuronal number by countering apoptosis in specific neuroblasts subpopulations, thus influencing final brain size and shape.
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References
Rakic, P. A small step for the cell, a giant leap for mankind: a hypothesis of neocortical expansion during evolution. Trends Neurosci. 18, 383–388 (1995).
Takahashi, T., Nowakowski, R.S. & Caviness, V.S.J. The mathematics of neocortical neuronogenesis. Dev. Neurosci. 19, 17–22 (1997).
Caviness, V.S.J., Takahashi, T. & Nowakowski, R.S. Numbers, time and neocortical neuronogenesis: a general developmental and evolutionary model. Trends Neurosci. 18, 379–383 (1995).
Haydar, T.F., Kuan, C-Y., Flavell, R.A. & Rakic, P. The role of cell death in regulating the size and shape of the mammalian forebrain. Cereb. Cortex 9, 621–626 (1999).
Kuan, C.-H., Roth, K.A., Flavell, R.A. & Rakic, P. Mechanisms of programmed cell death in the developing brain. Trends Neurosci. 23, 291–297 (2000).
de la Rosa, E.J. & de Pablo, F. Cell death in early neural development: beyond the neurotrophic theory. Trends Neurosci. 23, 454–458 (2000).
Pompeiano, M., Blaschke, A.J., Flavell, R.A., Srinivasan, A. & Chun, J. Decreased apoptosis in proliferative and postmitotic regions of the caspase 3-deficient embryonic central nervous system. J. Comp. Neurol. 423, 1–12 (2000).
Blaschke, A.J., Staley, K. & Chun, J. Widespread programmed cell death in proliferative and postmitotic regions of the fetal cerebral cortex. Development 122, 1165–1174 (1996).
Thomaidou, D., Mione, M.C., Cavanagh, J.F.R. & Parnavelas, J.G. Apoptosis and its relation to the cell cycle in the developing cerebral cortex. J. Neurosci. 17, 1075–1085 (1997).
Blaschke, A.J., Weiner, J.A. & Chun, J. Programmed cell death is a universal feature of embryonic and postnatal neuroproliferative regions throughout the central nervous system. J. Comp. Neurol. 396, 39–50 (1998).
Kuida, K. et al. Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice. Nature 384, 368–372 (1996).
Kuida, K. et al. Reduced apoptosis and cytochrome c-mediated caspase activation in mice lacking caspase 9. Cell 94, 325–337 (1998).
Hakem, R. et al. Differential requirement for caspase 9 in apoptotic pathways in vivo. Cell 94, 339–352 (1998).
Cecconi, F., Alvarez-Bolado, G., Meyer, B.I., Roth, K.A. & Gruss, P. Apaf1 (CED-4 Homolog) regulates programmed cell death in mammalian development. Cell 94, 727–737 (1998).
Yoshida, H. et al. Apaf1 is required for mitochondrial pathways of apoptosis and brain development. Cell 94, 739–750 (1998).
Nedivi, E., Hevroni, D., Naot, D., Israeli, D. & Citri, Y. Numerous candidate plasticity-related genes revealed by differential cDNA cloning. Nature 363, 718–722 (1993).
Hevroni, D. et al. Hippocampal plasticity involves extensive gene induction and multiple cellular mechanisms. J. Mol. Neurosci. 10, 75–98 (1998).
Naeve, G.S. et al. Neuritin: a gene induced by neural activity and neurotrophins that promotes neuritogenesis. Proc. Natl Acad. Sci. USA 94, 2648–2653 (1997).
Nedivi, E., Wu, G.Y. & Cline, H.T. Promotion of dendritic growth by CPG15, an activity-induced signaling molecule. Science 281, 1863–1866 (1998).
Cantallops, I., Haas, K. & Cline, H.T. Postsynaptic CPG15 promotes synaptic maturation and presynaptic axon arbor elaboration in vivo. Nat. Neurosci. 3, 1004–1011 (2000).
Corriveau, R., Shatz, C.J. & Nedivi, E. Dynamic regulation of cpg15 during activity-dependent synaptic development in the mammalian visual system. J. Neurosci. 19, 7999–8008 (1999).
Lee, W.C.A. & Nedivi, E. Extended plasticity of visual cortex in dark-reared animals may result from prolonged expression of genes like cpg15. J. Neurosci. 22, 1807–1815 (2002).
Hooper, N.M. Determination of glycosyl-phosphatidylinositol membrane protein anchorage. Proteomics 1, 748–755 (2001).
Lois, C., Hong, E.J., Pease, S., Brown, E.J. & Baltimore, D. Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 295, 868–872 (2002).
Rubinson, D.A. et al. A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nat. Genet. 33, 401–406 (2003).
McManus, M.T. & Sharp, P.A. Gene silencing in mammals by small interfering RNAs. Nat. Rev. Genet. 3, 737–747 (2002).
Chenn, A. & Walsh, C.A. Increased neuronal production, enlarged forebrains and cytoarchitectural distortions in β-catenin overexpressing transgenic mice. Cereb. Cortex 13, 599–606 (2003).
Motoyama, N. et al. Massive cell death of immature hematopoietic cells and neurons in Bcl-x-deficient mice. Science 267, 1506–1510 (1995).
Roth, K.A. et al. Epistatic and independent functions of Caspase-3 and Bcl-XL in developmental programmed cell death. Proc. Natl Acad. Sci. USA 97, 466–471 (2000).
Shindler, K.S., Latham, C.B. & Roth, K.A. bax deficiency prevents the increased cell death of immature neurons in bcl-x-deficient mice. J. Neurosci. 17, 3112–3119 (1997).
Knudson, C.M., Tung, K.S.K., Tourtellotte, W.G., Brown, G.A.J. & Korsmeyer, S.J. Bax-deficient mice with lymphoid hyperplasia and male germ cell death. Science 270, 96–99 (1995).
White, F.A., Keller-Peck, C.R., Knudson, C.M., Korsmeyer, S.J. & Snider, W.D. Widespread elimination of naturally occurring neuronal death in Bax-deficient mice. J. Neurosci. 18, 1428–1439 (1998).
Chenn, A. & Walsh, C.A. Regulation of cerebral cortical size by control of cell cycle exit in neural precursors. Science 297, 365–369 (2002).
Ortega, S., Ittmann, M., Tsang, S.H., Ehrlich, M. & Basilico, C. Neuronal defects and delayed wound healing in mice lacking fibroblast growth factor 2. Proc. Natl Acad. Sci. USA 95, 5672–5677 (1998).
Dono, R., Texido, G., Dussel, R., Ehmke, H. & Zeller, R. Impaired cerebral cortex development and blood pressure regulation in FGF-2-deficient mice. EMBO J. 17, 4213–4225 (1998).
Vaccarino, F.M. et al. Changes in cerebral cortex size are governed by fibroblast growth factor during embryogenesis. Nat. Neurosci. 2, 246–253 (1999).
Suh, J., Lu, N., Nicot, A., Tatsuno, I. & DiCicco-Bloom, E. PACAP is an anti-mitogenic signal in developing cerebral cortex. Nat. Neurosci. 4, 123–124 (2001).
Kingsbury, M.A., Rehen, S.K. & Contos, J.J.A. Higgins, C.M.a. & Chun, J. Non-proliferative effects of lysophosphatidic acid enhance cortical growth and folding. Nat. Neurosci. 6, 1292–1299 (2003).
Barnabé-Heider, F. & Miller, F.D. Endogenously produced neurotrophins regulate survival and differentiation of cortical progenitors via distinct signaling pathways. J. Neurosci. 23, 5149–5160 (2003).
Brunstrom, J.E., Gray-Swain, M.R., Osborne, P.A. & Pearlman, A.L. Neuronal heterotopias in the developing cerebral cortex produced by neurotrophin-4. Neuron 18, 505–517 (1997).
Ernfors, P., Merlio, J.-P. & Persson, H. Cells expressing mRNA for neurotrophins and their receptors during embryonic rat development. Eur. J. Neurosci. 4, 1140–1158 (1992).
Conover, J.C. & Yancopoulos, G.D. Neurotrophin regulation of the developing nervous system: analyses of knockout mice. Rev. Neurosci. 8, 13–27 (1997).
Götz, M. Doublecortin finds its place. Nat. Neurosci. 6, 1245–1247 (2003).
Corbo, J.C. et al. Doublecortin is required in mice for lamination of the hippocampus but not the neocortex. J. Neurosci. 22, 7548–7557 (2002).
Bai, J. et al. RNAi reveals doublecortin is required for radial migration in rat neocortex. Nat. Neurosci. 6, 1277–1282 (2003).
Gleeson, J.G. et al. Doublecortin, a brain-specific gene mutated in human X-linked lissencephaly and double cortex syndrome, encodes a putative signaling protein. Cell 92, 63–72 (1998).
Zhou, J. & Tang, X.C. Huperzine A attenuates apoptosis and mitochondria-dependent caspase-3 in rat cortical neurons. FEBS Lett. 526, 21–25 (2002).
Ghosh, A. & Greenberg, M.E. Distinct roles for bFGF and NT-3 in the regulation of cortical neurogenesis. Neuron 15, 89–103 (1995).
Walsh, C. & Cepko, C.L. Widespread dispersion of neuronal clones across functional regions of the cerebral cortex. Science 255, 434–440 (1992).
Sambrook, J., Fritsch, E.F. & Maniatis, T. Molecular Cloning: A Laboratory Manual. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989).
Acknowledgements
We thank members of the Nedivi laboratory and P. Garrity, J. Hoch, and J. Mintern for helpful comments on the manuscript, J. Cottrell for initiating and help with shRNA cloning and testing, C. Lois for advice on construction and use of lentivirus vectors, C. Walsh and E. Olson for guidance on embryonic injections, and J. Pungor for help with cell counts. This work was sponsored by grants from National Eye Institute and the Ellison Medical Foundation to E. Nedivi. U. Putz was supported by a postdoctoral fellowship from the Deutsche Forschungsgemeinschaft, and C. Harwell by a Ford Foundation predoctoral fellowship.
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Putz, U., Harwell, C. & Nedivi, E. Soluble CPG15 expressed during early development rescues cortical progenitors from apoptosis. Nat Neurosci 8, 322–331 (2005). https://doi.org/10.1038/nn1407
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DOI: https://doi.org/10.1038/nn1407
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