Vascular Endothelial Growth Factor Has Neurotrophic Activity and Stimulates Axonal Outgrowth, Enhancing Cell Survival and Schwann Cell Proliferation in the Peripheral Nervous System

Vascular endothelial growth factor (VEGF) is a mitogen for endothelial cells, and it promotes angiogenesis in vivo. Here we report that VEGF165 has neurotrophic actions on cultured adult mouse superior cervical ganglia (SCG) and dorsal root ganglia (DRG), measured as axonal outgrowth. Maximal effect was observed at 10–50 ng/ml for SCG and 100 ng/ml for DRG. VEGF-induced axonal outgrowth was inhibited by the mitogen-activated protein kinase kinase inhibitor PD 98059 but not by the protein kinase inhibitor K252a. VEGF also increased survival of both neurons and satellite cells and the number of proliferating Schwann cells. Immunocytochemistry and immunoblotting revealed that VEGF was expressed in virtually all nerve cells in the SCG but only in a population of small-diameter (<35 μm) neurons representing ∼30% of the neurons in DRG. Immunostaining showed that the VEGF receptor fetal liver kinase receptor (flk-1) was found on nerve cell bodies in DRG and to a lesser extent on neurons in SCG. Growth cones of regenerating axons from both types of ganglia exhibited flk-1 immunoreactivity, as did Schwann cells. We conclude that VEGF has both neurotrophic and mitogenic activity on cells in the peripheral nervous system.

[1]  V. Hamburger,et al.  In vitro experiments on the effects of mouse sarcomas 180 and 37 on the spinal and sympathetic ganglia of the chick embryo. , 1954, Cancer research.

[2]  H. Hydén,et al.  A glial protein specific for the nervous system. , 1966, Proceedings of the National Academy of Sciences of the United States of America.

[3]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[4]  E. Fenton.  Tissue culture assay of nerve growth factor and of the specific antiserum. , 1970, Experimental cell research.

[5]  G. Edelman,et al.  Effects of fasciculation on the outgrowth of neurites from spinal ganglia in culture , 1980, The Journal of cell biology.

[6]  M. Melamed,et al.  Detection of 5-bromodeoxyuridine (BrdUrd) incorporation by monoclonal antibodies: role of the DNA denaturation step. , 1985, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[7]  R. Pagano,et al.  A vital stain for the Golgi apparatus. , 1985, Science.

[8]  R. Rydel,et al.  Acidic and basic fibroblast growth factors promote stable neurite outgrowth and neuronal differentiation in cultures of PC12 cells , 1987, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[9]  D. Connolly,et al.  Vascular permeability factor, an endothelial cell mitogen related to PDGF. , 1989, Science.

[10]  P. Stroobant,et al.  Platelet-derived growth factors and fibroblast growth factors are mitogens for rat Schwann cells , 1990, The Journal of cell biology.

[11]  C. Heldin,et al.  Neurotrophic activity of platelet-derived growth factor (PDGF): Rat neuronal cells possess functional PDGF beta-type receptors and respond to PDGF. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[12]  G. Breier,et al.  Expression of vascular endothelial growth factor during embryonic angiogenesis and endothelial cell differentiation. , 1992, Development.

[13]  D. Dimitrov,et al.  Identification of the KDR tyrosine kinase as a receptor for vascular endothelial cell growth factor. , 1992, Biochemical and biophysical research communications.

[14]  A. Saltiel,et al.  Inhibition of the cellular actions of nerve growth factor by staurosporine and K252A results from the attenuation of the activity of the trk tyrosine kinase. , 1992, Biochemistry.

[15]  H Ueno,et al.  The fms-like tyrosine kinase, a receptor for vascular endothelial growth factor. , 1992, Science.

[16]  H. Dvorak,et al.  Vascular permeability factor (vascular endothelial growth factor) gene is expressed differentially in normal tissues, macrophages, and tumors. , 1992, Molecular biology of the cell.

[17]  N. Ferrara,et al.  Molecular and biological properties of the vascular endothelial growth factor family of proteins. , 1992, Endocrine reviews.

[18]  T. Quinn,et al.  Fetal liver kinase 1 is a receptor for vascular endothelial growth factor and is selectively expressed in vascular endothelium. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[19]  L. Williams,et al.  Vascular endothelial growth factor receptor expression during embryogenesis and tissue repair suggests a role in endothelial differentiation and blood vessel growth. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[20]  M. Shibuya,et al.  Different signal transduction properties of KDR and Flt1, two receptors for vascular endothelial growth factor. , 1994, The Journal of biological chemistry.

[21]  S. McMahon,et al.  Immunocytochemical Localization of trkA Receptors in Chemically Identified Subgroups of Adult Rat Sensory Neurons , 1995, The European journal of neuroscience.

[22]  Janet Rossant,et al.  Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice , 1995, Nature.

[23]  G. Yancopoulos,et al.  A BDNF autocrine loop in adult sensory neurons prevents cell death , 1995, Nature.

[24]  J. Stone,et al.  Development of retinal vasculature is mediated by hypoxia-induced vascular endothelial growth factor (VEGF) expression by neuroglia , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[25]  M. Merrill,et al.  Differential expression of vascular endothelial growth factor (vascular permeability factor) forms in rat tissues. , 1995, Growth factors.

[26]  Per A. R. Ekström,et al.  Neurones and glial cells of the mouse sciatic nerve undergo apoptosis after injury in vivo and in vitro , 1995, Neuroreport.

[27]  J. Rossant,et al.  Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium , 1995, Nature.

[28]  A. Fagan,et al.  TrkA, But Not TrkC, Receptors Are Essential for Survival of Sympathetic Neurons In Vivo , 1996, The Journal of Neuroscience.

[29]  I. Fariñas,et al.  A Reciprocal Cell–Cell Interaction Mediated by NT-3 and Neuregulins Controls the Early Survival and Development of Sympathetic Neuroblasts , 1996, Neuron.

[30]  D. Tonge,et al.  Axonal outgrowth and neuronal apoptosis in cultured adult mouse dorsal root ganglion preparations: effects of neurotrophins, of inhibition of neurotrophin actions and of prior axotomy , 1996, Neuroscience.

[31]  N. Ferrara,et al.  The biology of vascular endothelial growth factor. , 1997, Endocrine reviews.

[32]  M. Schwab,et al.  VEGF mRNA Induction Correlates With Changes in the Vascular Architecture Upon Spinal Cord Damage in the Rat , 1997, The European journal of neuroscience.

[33]  A. Windebank,et al.  Inhibition of axonal growth from sensory neurons by excess nerve growth factor , 1997, Annals of neurology.

[34]  A. Mizoguchi,et al.  Basic Fibroblast Growth Factor Promotes Extension of Regenerating Axons of Peripheral Nerve. In Vivo Experiments Using a Schwann Cell Basal Lamina Tube Model , 1997, Journal of neurocytology.

[35]  Mariann Sondell,et al.  The insulin‐like growth factors I and II stimulate proliferation of different types of Schwann cells , 1997, Neuroreport.

[36]  T. Yagi,et al.  Neuropilin–Semaphorin III/D-Mediated Chemorepulsive Signals Play a Crucial Role in Peripheral Nerve Projection in Mice , 1997, Neuron.

[37]  J. Waltenberger,et al.  The Vascular Endothelial Growth Factor Receptor KDR Activates Multiple Signal Transduction Pathways in Porcine Aortic Endothelial Cells* , 1997, The Journal of Biological Chemistry.

[38]  G. Neufeld,et al.  VEGF145, a Secreted Vascular Endothelial Growth Factor Isoform That Binds to Extracellular Matrix* , 1997, The Journal of Biological Chemistry.

[39]  G. Terenghi,et al.  Effects of delayed re‐innervation on the expression of c‐erbB receptors by chronically denervated rat Schwann cells in vivo , 1997, Glia.

[40]  S. Kaul,et al.  A Novel Alternatively Spliced Form of Murine Vascular Endothelial Growth Factor, VEGF 115* , 1998, The Journal of Biological Chemistry.

[41]  Shay Soker,et al.  Neuropilin-1 Is Expressed by Endothelial and Tumor Cells as an Isoform-Specific Receptor for Vascular Endothelial Growth Factor , 1998, Cell.

[42]  D. Tonge,et al.  Use of explant cultures of peripheral nerves of adult vertebrates to study axonal regeneration in vitro , 1998, Progress in Neurobiology.