Doublecortin expression levels in adult brain reflect neurogenesis

Progress in the field of neurogenesis is currently limited by the lack of tools enabling fast and quantitative analysis of neurogenesis in the adult brain. Doublecortin (DCX) has recently been used as a marker for neurogenesis. However, it was not clear whether DCX could be used to assess modulations occurring in the rate of neurogenesis in the adult mammalian central nervous system following lesioning or stimulatory factors. Using two paradigms increasing neurogenesis levels (physical activity and epileptic seizures), we demonstrate that quantification of DCX‐expressing cells allows for an accurate measurement of modulations in the rate of adult neurogenesis. Importantly, we excluded induction of DCX expression during physiological or reactive gliogenesis and excluded also DCX re‐expression during regenerative axonal growth. Our data validate DCX as a reliable and specific marker that reflects levels of adult neurogenesis and its modulation. We demonstrate that DCX is a valuable alternative to techniques currently used to measure the levels of neurogenesis. Importantly, in contrast to conventional techniques, analysis of neurogenesis through the detection of DCX does not require in vivo labelling of proliferating cells, thereby opening new avenues for the study of human neurogenesis under normal and pathological conditions.

[1]  J. Hinds,et al.  Neurogenesis in the adult rat: electron microscopic analysis of light radioautographs. , 1977, Science.

[2]  U. Lendahl,et al.  Nestin expression is lost in a neural stem cell line through a mechanism involving the proteasome and Notch signalling. , 2004, Brain research. Developmental brain research.

[3]  Y. Berwald‐Netter,et al.  A Novel CNS Gene Required for Neuronal Migration and Involved in X-Linked Subcortical Laminar Heterotopia and Lissencephaly Syndrome , 1998, Cell.

[4]  B. McEwen,et al.  Doublecortin expression in the adult rat telencephalon , 2001, The European journal of neuroscience.

[5]  P. Popovich,et al.  Rats and mice exhibit distinct inflammatory reactions after spinal cord injury , 2003, The Journal of comparative neurology.

[6]  J. Kapfhammer,et al.  Overexpression of the neural growth-associated protein GAP-43 induces nerve sprouting in the adult nervous system of transgenic mice , 1995, Cell.

[7]  Q. Zhu,et al.  Protective effect of neurofilament heavy gene overexpression in motor neuron disease induced by mutant superoxide dismutase. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[8]  C. Lois,et al.  Long-distance neuronal migration in the adult mammalian brain. , 1994, Science.

[9]  D. Geschwind,et al.  Dentate Granule Cell Neurogenesis Is Increased by Seizures and Contributes to Aberrant Network Reorganization in the Adult Rat Hippocampus , 1997, The Journal of Neuroscience.

[10]  R. McKay,et al.  CNS stem cells express a new class of intermediate filament protein , 1990, Cell.

[11]  Alexander Sasha Rabchevsky,et al.  Experimental modeling of spinal cord injury: characterization of a force-defined injury device. , 2003, Journal of neurotrauma.

[12]  F. Gage,et al.  Epidermal Growth Factor and Fibroblast Growth Factor-2 Have Different Effects on Neural Progenitors in the Adult Rat Brain , 1997, The Journal of Neuroscience.

[13]  J. Winkler,et al.  High Efficacy of Clonal Growth and Expansion of Adult Neural Stem Cells , 2003, Laboratory Investigation.

[14]  Hirofumi Nakatomi,et al.  Regeneration of Hippocampal Pyramidal Neurons after Ischemic Brain Injury by Recruitment of Endogenous Neural Progenitors , 2002, Cell.

[15]  H. Scharfman,et al.  Granule-Like Neurons at the Hilar/CA3 Border after Status Epilepticus and Their Synchrony with Area CA3 Pyramidal Cells: Functional Implications of Seizure-Induced Neurogenesis , 2000, The Journal of Neuroscience.

[16]  F. Gage,et al.  Running increases cell proliferation and neurogenesis in the adult mouse dentate gyrus , 1999, Nature Neuroscience.

[17]  O. Lindvall,et al.  Caspase inhibitors increase short‐term survival of progenitor‐cell progeny in the adult rat dentate gyrus following status epilepticus , 2001, The European journal of neuroscience.

[18]  P. Rakic,et al.  Three‐dimensional counting: An accurate and direct method to estimate numbers of cells in sectioned material , 1988, The Journal of comparative neurology.

[19]  J. García-Verdugo,et al.  Astrocytes Give Rise to New Neurons in the Adult Mammalian Hippocampus , 2001, The Journal of Neuroscience.

[20]  G. Kempermann Regulation of adult hippocampal neurogenesis - implications for novel theories of major depression. , 2002, Bipolar disorders.

[21]  T. Palmer,et al.  Fibroblast Growth Factor-2 Activates a Latent Neurogenic Program in Neural Stem Cells from Diverse Regions of the Adult CNS , 1999, The Journal of Neuroscience.

[22]  S. Bayer,et al.  Changes in the total number of dentate granule cells in juvenile and adult rats: A correlated volumetric and 3H-thymidine autoradiographic study , 2004, Experimental Brain Research.

[23]  Alan Carleton,et al.  Becoming a new neuron in the adult olfactory bulb , 2003, Nature Neuroscience.

[24]  T J Sejnowski,et al.  Running enhances neurogenesis, learning, and long-term potentiation in mice. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Jun Ding,et al.  Different regulatory elements are necessary for αI tubulin induction during CNS development and regeneration , 2000, Neuroreport.

[26]  D. Steindler,et al.  Cell and molecular analysis of the developing and adult mouse subventricular zone of the cerebral hemispheres , 1995, The Journal of comparative neurology.

[27]  J. Winkler,et al.  Analysis of neurogenesis and programmed cell death reveals a self-renewing capacity in the adult rat brain , 2000, Neuroscience Letters.

[28]  Jürgen Winkler,et al.  Long‐term survival and cell death of newly generated neurons in the adult rat olfactory bulb , 2002, The European journal of neuroscience.

[29]  O. Lindvall,et al.  Neuronal replacement from endogenous precursors in the adult brain after stroke , 2002, Nature Medicine.

[30]  M. Holmes,et al.  Behavioral feedback regulation of circadian rhythm phase angle in light-dark entrained mice. , 2000, American journal of physiology. Regulatory, integrative and comparative physiology.

[31]  F. Gage,et al.  Functional neurogenesis in the adult hippocampus , 2002, Nature.

[32]  Lin Xie,et al.  Increased hippocampal neurogenesis in Alzheimer's disease , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[33]  F. Gage,et al.  Neurogenesis in the adult human hippocampus , 1998, Nature Medicine.

[34]  G. Kempermann,et al.  Subpopulations of proliferating cells of the adult hippocampus respond differently to physiologic neurogenic stimuli , 2003, The Journal of comparative neurology.

[35]  C. Cooper-Kuhn,et al.  Is it all DNA repair? Methodological considerations for detecting neurogenesis in the adult brain. , 2002, Brain research. Developmental brain research.

[36]  Daniel A. Lim,et al.  Subventricular Zone Astrocytes Are Neural Stem Cells in the Adult Mammalian Brain , 1999, Cell.

[37]  J. Winkler,et al.  Molecular mechanisms of neuronal migration disorders, quo vadis? , 2001, Current molecular medicine.

[38]  P. Hof,et al.  Overexpression of Wild Type But Not an FAD Mutant Presenilin-1 Promotes Neurogenesis in the Hippocampus of Adult Mice , 2002, Neurobiology of Disease.

[39]  Q. Zhu,et al.  Disruption of Type IV Intermediate Filament Network in Mice Lacking the Neurofilament Medium and Heavy Subunits , 1999, Journal of neurochemistry.

[40]  J. Altman,et al.  Autoradiographic and histological evidence of postnatal hippocampal neurogenesis in rats , 1965, The Journal of comparative neurology.

[41]  F. Gage,et al.  Enriched environment and physical activity stimulate hippocampal but not olfactory bulb neurogenesis , 2003, The European journal of neuroscience.

[42]  Mitchel S. Berger,et al.  Unique astrocyte ribbon in adult human brain contains neural stem cells but lacks chain migration , 2004, Nature.

[43]  P. Caroni,et al.  Shared and Unique Roles of Cap23 and Gap43 in Actin Regulation, Neurite Outgrowth, and Anatomical Plasticity , 2000, The Journal of cell biology.

[44]  O. V. Stepanenko,et al.  High stability of Discosoma DsRed as compared to Aequorea EGFP. , 2003, Biochemistry.

[45]  Jürgen Winkler,et al.  Transient expression of doublecortin during adult neurogenesis , 2003, The Journal of comparative neurology.

[46]  Blair R. Leavitt,et al.  Induction of neurogenesis in the neocortex of adult mice , 2000, Nature.

[47]  H. Cameron,et al.  Short‐term and long‐term survival of new neurons in the rat dentate gyrus , 2003, The Journal of comparative neurology.

[48]  M. Mattson,et al.  Disruption of neurogenesis by amyloid β‐peptide, and perturbed neural progenitor cell homeostasis, in models of Alzheimer's disease , 2002, Journal of neurochemistry.

[49]  I. Scheffer,et al.  doublecortin , a Brain-Specific Gene Mutated in Human X-Linked Lissencephaly and Double Cortex Syndrome, Encodes a Putative Signaling Protein , 1998, Cell.

[50]  Kozo Nakamura,et al.  Proliferation of Parenchymal Neural Progenitors in Response to Injury in the Adult Rat Spinal Cord , 2001, Experimental Neurology.

[51]  F. Gage,et al.  Proliferation and Differentiation of Progenitor Cells Throughout the Intact Adult Rat Spinal Cord , 2000, The Journal of Neuroscience.

[52]  M Kokaia,et al.  Apoptosis and proliferation of dentate gyrus neurons after single and intermittent limbic seizures. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[53]  Arturo Alvarez-Buylla,et al.  Maturation and Death of Adult-Born Olfactory Bulb Granule Neurons: Role of Olfaction , 2002, The Journal of Neuroscience.

[54]  T. Palmer,et al.  Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation , 1996, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[55]  Maria B. Luskin,et al.  Restricted proliferation and migration of postnatally generated neurons derived from the forebrain subventricular zone , 1993, Neuron.

[56]  H. J. G. Gundersen,et al.  The new stereological tools: Disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis , 1988, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[57]  A. Shetty,et al.  Efficacy of doublecortin as a marker to analyse the absolute number anddendritic growth of newly generated neurons in the adult dentate gyrus , 2004, The European journal of neuroscience.

[58]  P. Caroni,et al.  Depletion of 43-kD growth-associated protein in primary sensory neurons leads to diminished formation and spreading of growth cones , 1993, The Journal of cell biology.

[59]  C. Lois,et al.  Proliferating subventricular zone cells in the adult mammalian forebrain can differentiate into neurons and glia. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[60]  J. Wojtowicz,et al.  Kindling-induced neurogenesis in the dentate gyrus of the rat , 1998, Neuroscience Letters.