Essential role of heparan sulfates in axon navigation and targeting in the developing visual system.

Heparan sulfate (HS) is abundant in the developing brain and is a required co-factor for many types of fibroblast growth factor (FGF) signaling in vitro. We report that some HSs, when added exogenously to the developing Xenopus optic pathway, severely disrupt target recognition causing axons from the retina to bypass their primary target, the optic tectum. Significantly, HS sidechains from a neuroepithelial perlecan variant that preferentially bind FGF-2, HS(FGF-2), cause aberrant targeting, whereas those that preferentially bind FGF-1 do not. Charge-matched fragments of HS(FGF-2) show that the mistargeting activity associates with the FGF-binding fragments. Heparitinase removal of native HSs at the beginning of optic tract formation retards retinal axon elongation; addition of FGF-2 restores axon extension but axons lose directionality. Late HS removal, after axons have extended through the tract, elicits a tectal bypass phenotype indicating a growth promoting and guidance function for native HSs. Our results demonstrate that different HS sidechains from the same core protein differentially affect axon growth in vivo, possibly due to their distinct FGF-binding preferences, and suggest that growth factors and HSs are important partners in regulating axon growth and guidance in the developing visual system.

[1]  C. Goodman,et al.  Ectopic expression of connectin reveals a repulsive function during growth cone guidance and synapse formation , 1994, Neuron.

[2]  Jonathan A. Raper,et al.  The enrichment of a neuronal growth cone collapsing activity from embryonic chick brain , 1990, Neuron.

[3]  O. Yoshie,et al.  A syngeneic monoclonal antibody to murine Meth-A sarcoma (HepSS-1) recognizes heparan sulfate glycosaminoglycan (HS-GAG): cell density and transformation dependent alteration in cell surface HS-GAG defined by HepSS-1. , 1986, Journal of immunology.

[4]  P. Dell’Era,et al.  Distinct role of 2-O-, N-, and 6-O-sulfate groups of heparin in the formation of the ternary complex with basic fibroblast growth factor and soluble FGF receptor-1. , 1994, Biochemical and biophysical research communications.

[5]  D. Gospodarowicz,et al.  Heparin protects basic and acidic FGF from inactivation , 1986, Journal of cellular physiology.

[6]  A. Loewy,et al.  Neuronal cell–cell adhesion depends on interactions of N-CAM with heparin-like molecules , 1986, Nature.

[7]  A. Lander,et al.  Inhibitors and Promoters of Thalamic Neuron Adhesion and Outgrowth in Embryonic Neocortex: Functional Association with Chondroitin Sulfate , 1996, Neuron.

[8]  B. Olwin,et al.  Activating and inhibitory heparin sequences for FGF-2 (basic FGF). Distinct requirements for FGF-1, FGF-2, and FGF-4. , 1993, The Journal of biological chemistry.

[9]  J. Xu,et al.  An essential heparin-binding domain in the fibroblast growth factor receptor kinase. , 1993, Science.

[10]  F. Matsui,et al.  Brain development and multiple molecular species of proteoglycan , 1994, Neuroscience Research.

[11]  J. Faber,et al.  Normal Table of Xenopus Laevis (Daudin) , 1958 .

[12]  C E Holt,et al.  A single-cell analysis of early retinal ganglion cell differentiation in Xenopus: from soma to axon tip , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[13]  Jeffrey D. Esko,et al.  Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor , 1991, Cell.

[14]  A. Yayon,et al.  Perlecan, basal lamina proteoglycan, promotes basic fibroblast growth factor-receptor binding, mitogenesis, and angiogenesis , 1994, Cell.

[15]  C. Holt,et al.  Navigational errors made by growth cones without filopodia in the embryonic xenopus brain , 1993, Neuron.

[16]  T. Maciag,et al.  The heparin-binding (fibroblast) growth factor family of proteins. , 1989, Annual review of biochemistry.

[17]  K Isahara,et al.  The interaction of vascular endothelial cells and dorsal root ganglion neurites is mediated by vitronectin and heparan sulfate proteoglycans. , 1995, Brain research. Developmental brain research.

[18]  H. Yost,et al.  Embryonic expression patterns of Xenopus syndecans , 1996, Mechanisms of Development.

[19]  G. Waksman,et al.  FGF binding and FGF receptor activation by synthetic heparan-derived di- and trisaccharides. , 1995, Science.

[20]  H. Rauvala,et al.  Neurite Outgrowth in Brain Neurons Induced by Heparin-binding Growth-associated Molecule (HB-GAM) Depends on the Specific Interaction of HB-GAM with Heparan Sulfate at the Cell Surface (*) , 1996, The Journal of Biological Chemistry.

[21]  T. Jessell,et al.  The netrins define a family of axon outgrowth-promoting proteins homologous to C. elegans UNC-6 , 1994, Cell.

[22]  C. Holt,et al.  Retinal axons with and without their somata, growing to and arborizing in the tectum of Xenopus embryos: a time-lapse video study of single fibres in vivo. , 1987, Development.

[23]  A. Skubitz,et al.  Recognition of the A chain carboxy-terminal heparin binding region of fibronectin involves multiple sites: two contiguous sequences act independently to promote neural cell adhesion , 1990, The Journal of cell biology.

[24]  A. Lander.  Mechanisms by which molecules guide axons. , 1990, Current opinion in cell biology.

[25]  A. Lander,et al.  A diverse set of developmentally regulated proteoglycans is expressed in the rat central nervous system , 1990, Neuron.

[26]  J. Turnbull,et al.  Molecular organization of heparan sulphate from human skin fibroblasts. , 1990, The Biochemical journal.

[27]  V. Nurcombe,et al.  Developmental regulation of neural response to FGF-1 and FGF-2 by heparan sulfate proteoglycan. , 1993, Science.

[28]  J. Gerhart,et al.  Early cellular interactions promote embryonic axis formation in Xenopus laevis. , 1984, Developmental biology.

[29]  U. Greferath,et al.  A proteoglycan that activates fibroblast growth factors during early neuronal development is a perlecan variant. , 1996, Development.

[30]  J. Turnbull,et al.  Identification of the basic fibroblast growth factor binding sequence in fibroblast heparan sulfate. , 1992, The Journal of biological chemistry.

[31]  H. Hondermarck,et al.  Embryonic brain-derived heparan sulfate inhibits cellular membrane binding and biological activity of basic fibroblast growth factor. , 1992, Brain research. Developmental brain research.

[32]  C. Holt,et al.  FGF signaling and target recognition in the developing xenopus visual system , 1995, Neuron.

[33]  J. Turnbull,et al.  Heparan sulphate in the binding and activation of basic fibroblast growth factor. , 1992, Glycobiology.

[34]  D. Small,et al.  Heparan Sulfates Mediate the Binding of Basic Fibroblast Growth Factor to a Specific Receptor on Neural Precursor Cells (*) , 1995, The Journal of Biological Chemistry.

[35]  M. Kan,et al.  Alternately Spliced NH2-terminal Immunoglobulin-like Loop I in the Ectodomain of the Fibroblast Growth Factor (FGF) Receptor 1 Lowers Affinity for both Heparin and FGF-1(*) , 1995, The Journal of Biological Chemistry.

[36]  W. Halfter.  A heparan sulfate proteoglycan in developing avian axonal tracts , 1993, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[37]  M. Jaye,et al.  Heparin-induced oligomerization of FGF molecules is responsible for FGF receptor dimerization, activation, and cell proliferation , 1994, Cell.

[38]  A. Yayon,et al.  Engineering of fibroblast growth factor: alteration of receptor binding specificity. , 1995, Biochemistry.

[39]  C. Holt,et al.  Precocious pathfinding: Retinal axons can navigate in an axonless brain , 1992, Neuron.

[40]  W. Harris,et al.  Growth cone interactions with a glial cell line from embryonic Xenopus retina. , 1989, Developmental biology.

[41]  J. Wilting,et al.  Recent research on the biological activity of suramin. , 1993, Pharmacological reviews.

[42]  X. Bai,et al.  Developmental changes in heparan sulfate expression: in situ detection with mAbs , 1992, The Journal of cell biology.

[43]  V. Nurcombe,et al.  Co-localization of FGF-2 and a novel heparan sulphate proteoglycan in embryonic mouse brain. , 1994, NeuroReport.

[44]  Paul C. Letourneau,et al.  Cell-Surface Heparan Sulfate Proteoglycan From Embryonic Rat Spinal Cord , 2004 .

[45]  H. Keshishian,et al.  Fasciclin III as a synaptic target recognition molecule in Drosophila , 1995, Nature.

[46]  J. Gurdon.  Chapter 7 Methods for Nuclear Transplantation in Amphibia , 1977 .

[47]  W. Harris,et al.  Two cellular inductions involved in photoreceptor determination in the Xenopus retina , 1992, Neuron.

[48]  A. Brown,et al.  The proto‐oncogene int‐1 encodes a secreted protein associated with the extracellular matrix. , 1990, The EMBO journal.

[49]  D. Rifkin,et al.  Heparin increases the affinity of basic fibroblast growth factor for its receptor but is not required for binding. , 1994, The Journal of biological chemistry.

[50]  C. Holt,et al.  Inhibition of FGF Receptor Activity in Retinal Ganglion Cell Axons Causes Errors in Target Recognition , 1996, Neuron.

[51]  D. Edwards,et al.  Antibodies against filamentous components in discrete cell types of the mouse retina , 1984, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[52]  H. van den Berghe,et al.  Stimulation of fibroblast growth factor receptor-1 occupancy and signaling by cell surface-associated syndecans and glypican , 1996, The Journal of cell biology.

[53]  J. Gurdon.  Methods for nuclear transplantation in amphibia. , 1977, Methods in cell biology.

[54]  M. Goldfarb,et al.  Heparin can activate a receptor tyrosine kinase. , 1995, EMBO Journal.

[55]  J. Gurdon,et al.  Normal table of Xenopus laevis (Daudin) , 1995 .

[56]  P. Barr,et al.  The Molecular Biology of Heparan Sulfate Fibroblast Growth Factor Receptors a , 1991, Annals of the New York Academy of Sciences.

[57]  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.

[58]  I. Lax,et al.  Regulation of growth factor activation by proteoglycans: What is the role of the low affinity receptors? , 1995, Cell.

[59]  J. Denburg,et al.  A role for proteoglycans in the guidance of a subset of pioneer axons in cultured embryos of the cockroach , 1992, Neuron.

[60]  B. Olwin,et al.  Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation , 1991, Science.

[61]  S. Drake,et al.  A cell-surface heparan sulfate proteoglycan mediates neural cell adhesion and spreading on a defined sequence from the C-terminal cell and heparin binding domain of fibronectin, FN-C/H II , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[62]  R. Saxod,et al.  Influence of glycosaminoglycans on neurite morphology and outgrowth patterns in vitro , 1989, International Journal of Developmental Neuroscience.

[63]  M. Klagsbrun,et al.  A dual receptor system is required for basic fibroblast growth factor activity , 1991, Cell.

[64]  Paul C. Letourneau,et al.  Central and peripheral neurite outgrowth differs in preference for heparin-binding versus integrin-binding sequences , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.