Identification of pleiotrophin in conditioned medium secreted from neural stem cells by SELDI-TOF and SELDI-tandem mass spectrometry.

Neural stem cells (NSCs) are multipotential progenitor cells that have self-renewal activity. Since the fates of the NSCs in situ depend on their niche containing growth factors and cytokines, we performed surface enhanced laser desorption/ionization time-of flight mass spectrometry (SELDI-TOF-MS) to screen for differentially secreted proteins in conditioned medium of neural stem cells and compared with that of NIH3T3 cells. A 15.3-kDa protein detected only in the conditioned medium of neural stem cells was determined as pleiotrophin (PTN) by SELDI-TOF-MS and ProteinChip-tandem MS systems. Identification of pleiotrophin was further confirmed by one-dimensional SDS gel electrophoresis and Edman degradation analysis. The mRNA transcripts of PTN and its receptors [receptor protein tyrosine phosphatase (RPTP) beta/zeta, N-syndecan and anaplastic lymphoma kinase (ALK)] were detected in neurosphere, suggesting that pleiotrophin signaling systems are present in the neural stem cells and are involved in the modulation of fate of neural stem cells.

[1]  S. Weinberger,et al.  Recent trends in protein biochip technology. , 2000, Pharmacogenomics.

[2]  D. Carey,et al.  Isolation of a neuronal cell surface receptor of heparin binding growth-associated molecule (HB-GAM). Identification as N-syndecan (syndecan-3). , 1994, The Journal of biological chemistry.

[3]  S. Temple,et al.  A self-renewing multipotential stem cell in embryonic rat cerebral cortex , 1994, Nature.

[4]  C. Powers,et al.  Pleiotrophin Signaling through Anaplastic Lymphoma Kinase Is Rate-limiting for Glioblastoma Growth* , 2002, The Journal of Biological Chemistry.

[5]  T. Deuel,et al.  A novel 17 kD heparin-binding growth factor (HBGF-8) in bovine uterus: purification and N-terminal amino acid sequence. , 1989, Biochemical and biophysical research communications.

[6]  Robert H Miller,et al.  Sonic hedgehog-induced neural precursor proliferation after adult rodent spinal cord injury. , 2003, Journal of neurosurgery.

[7]  M. Watson,et al.  Cloning and expression of a developmentally regulated protein that induces mitogenic and neurite outgrowth activity. , 1990, Science.

[8]  H. Rauvala,et al.  N‐Syndecan and HB‐GAM (Heparin‐Binding Growth‐Associated Molecule) associate with early axonal tracts in the rat brain , 1998, The European journal of neuroscience.

[9]  Volkmar Lessmann,et al.  Neurotrophin secretion: current facts and future prospects , 2003, Progress in Neurobiology.

[10]  B. Martin,et al.  Identification of Collagen-Binding Proteins inLactobacillus spp. with Surface-Enhanced Laser Desorption/Ionization–Time of Flight ProteinChip Technology , 2000, Applied and Environmental Microbiology.

[11]  A. Martínez-Serrano,et al.  Human neural stem and progenitor cells: in vitro and in vivo properties, and potential for gene therapy and cell replacement in the CNS. , 2001, Current gene therapy.

[12]  W. Snider,et al.  Localization of pleiotrophin and its mRNA in subpopulations of neurons and their corresponding axonal tracts suggests important roles in neural-glial interactions during development and in maturity. , 1996, Journal of neurobiology.

[13]  M. Rao,et al.  Gliogenesis in the central nervous system , 2000, Glia.

[14]  M. Kaksonen,et al.  Co-expression of heparin-binding growth-associated molecule (HB-GAM) and N-syndecan (syndecan-3) in developing rat brain , 1995, Neuroscience Letters.

[15]  F M Watt,et al.  Out of Eden: stem cells and their niches. , 2000, Science.

[16]  K. Hobson,et al.  Neuroepithelial stem cells from the embryonic spinal cord: isolation, characterization, and clonal analysis. , 1997, Developmental biology.

[17]  R. Cappai,et al.  Interactions between Fibroblast Growth Factors and Notch Regulate Neuronal Differentiation , 2001, The Journal of Neuroscience.

[18]  Hideyuki Okano,et al.  Neurogenesis by Progenitor Cells in the Ischemic Adult Rat Hippocampus , 2001, Stroke.

[19]  T. Shintani,et al.  6B4 Proteoglycan/Phosphacan, an Extracellular Variant of Receptor-like Protein-tyrosine Phosphatase ζ/RPTPβ, Binds Pleiotrophin/Heparin-binding Growth-associated Molecule (HB-GAM)* , 1996, The Journal of Biological Chemistry.

[20]  S. Weinberger,et al.  Recent advancements in surface‐enhanced laser desorption/ionization‐time of flight‐mass spectrometry , 2000, Electrophoresis.

[21]  J. Vanderhaeghen,et al.  Cellular distribution of the new growth factor Pleiotrophin (HB-GAM) mRNA in developing and adult rat tissues , 1992, Anatomy and Embryology.

[22]  K. Narasimhan Signaling myopia , 1999, Nature Neuroscience.

[23]  K. Matsumoto,et al.  A novel family of heparin-binding growth factors, pleiotrophin and midkine, is expressed in the developing rat cerebral cortex. , 1994, Brain research. Developmental brain research.

[24]  F. Gage Discussion point Stem cells of the central nervous system , 1998, Current Opinion in Neurobiology.

[25]  M. Schachner,et al.  High Affinity Binding and Overlapping Localization of Neurocan and Phosphacan/Protein-tyrosine Phosphatase-ζ/β with Tenascin-R, Amphoterin, and the Heparin-binding Growth-associated Molecule* , 1998, The Journal of Biological Chemistry.

[26]  S. Nigam,et al.  Identification of pleiotrophin as a mesenchymal factor involved in ureteric bud branching morphogenesis. , 2001, Development.

[27]  H. Rauvala An 18‐kd heparin‐binding protein of developing brain that is distinct from fibroblast growth factors. , 1989, The EMBO journal.

[28]  E. Parati,et al.  Epidermal and Fibroblast Growth Factors Behave as Mitogenic Regulators for a Single Multipotent Stem Cell-Like Population from the Subventricular Region of the Adult Mouse Forebrain , 1999, The Journal of Neuroscience.

[29]  I. J. Mehr Preparing for the revolution--pharmacogenomics and the clinical lab. , 2000, Pharmacogenomics.

[30]  David J. Anderson,et al.  Regulatory Mechanisms in Stem Cell Biology , 1997, Cell.

[31]  R. McKay,et al.  Stem Cells in the Central Nervous System , 1997, Science.

[32]  D. Anderson,et al.  Stem Cells and Pattern Formation in the Nervous System The Possible versus the Actual , 2001, Neuron.

[33]  J. Leckman,et al.  Basic Fibroblast Growth Factor (Fgf2) Is Necessary for Cell Proliferation and Neurogenesis in the Developing Cerebral Cortex , 2000, The Journal of Neuroscience.

[34]  J. Milbrandt,et al.  Developmentally regulated expression of pleiotrophin, a novel heparin binding growth factor, in the nervous system of the rat. , 1993, Brain research. Developmental brain research.

[35]  F. Gage,et al.  Mammalian neural stem cells. , 2000, Science.

[36]  Clive N. Svendsen,et al.  New prospects for human stem-cell therapy in the nervous system , 1999, Trends in Neurosciences.