Abstract
In the central and peripheral nervous systems a heterogeneous group of proteins constituting the thrombospondin superfamily provides a cue for axonal pathfinding. They either contain or are devoid of the tripeptide RGD, and the sequence(s) and mechanism(s) which trigger in vitro their neurite-promoting activity have remained unclear. In this study, we reconsider the problem of whether sequences present in the thrombospondin type 1 repeats (TSRs), and independent of the well-known RGD-binding site, may activate integrins and account for their neurite-promoting activity. SCO-spondin is a newly identified member of the thrombospondin superfamily, which shows a multidomain organization with a great number of TSR motifs but no RGD sequence. Previous research has implicated oligopeptides derived from SCO-spondin TSRs in in-vitro development of various neuronal cell types. In this study, we investigate whether function-blocking antibodies directed against integrin subunits can block these effects in cell line B104, cloned from a neuroblastoma of the rat central nervous system. By two different approaches: flow cytometry revealing short-term effects and cell cultures revealing long-term effects, we show that: (a) activation of cell metabolism, (b) changes in cell size and structure, and (c) neurite-promoting activity induced by TSR oligopeptides are inhibited by function-blocking antibodies to β1-subunit. Using a panel of function-blocking antibodies directed against various integrin α-subunits we show that the α1-subunit might be the partner of the β1-subunit in B104 cells. Thus, we demonstrate that an original sequence within a TSR motif from SCO-spondin promotes neurite outgrowth through an intracellular signal driven by integrins, independently of an RGD-binding site.
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References
Adams JC, Tucker RP (2000) The thrombospondin type 1 repeat (TSR) superfamily: diverse proteins with related roles in neuronal development. Dev Dyn 218:280–299
Adams RH, Beets H, Püschel AW (1996) A novel class of murine semaphorins with homology to thrombospondin is differentially expressed during early embryogenesis. Mech Dev 57:33–45
Brown EJ, Frazier WA (2001) Integrin-associated protein (CD47) and its ligands. Trends Cell Biol 11:130–135
Burstyn-Cohen T, Frumkin A, Xu YT, Sherer SS, Klar A (1998) Accumulation of F-spondin in injured peripheral nerve promotes the outgrowth of sensory axons. J Neurosci 18:8875–8885
Carmeliet G, Himpens B, Cassiman J (1994) Selective increase in the binding of the α1β1 integrin for collagen type IV during neurite outgrowth of human neuroblastoma TR 14 cells. J Cell Sci 107:3379–3392
Chen H, Herndon ME, Lawler J (2000) The cell biology of thrombospondin-1. Matrix Biol 19:597–614
Cook G, Tannahil D, Keynes RJ (1998) Axon guidance to and from choice points. Curr Opin Neurobiol 8:64–72
Creveaux I, Gobron S, Meiniel R, Dastugue B, Meiniel A (1998) Complex expression pattern of the SCO-spondin gene in the bovine subcommissural organ: toward an explanation for Reissner’s fiber complexity. Mol Brain Res 55:45–53
Debby-Brafman A, Burstyn-Cohen T, Klar A, Kalcheim C (1999) F-spondin, expressed in somite regions avoided by neural crest cells, mediates inhibition of distinct somite domains to neural crest migration. Neuron 22:475–488
DeFreitas MF, Yoshida CK, Frazier WA, Mendrick DL, Kypta RM, Reichardt LF (1995) Identification of integrin α3β1 as a neuronal thrombospondin receptor mediating neurite outgrowth. Neuron 15:333–343
El Bitar F, Bamdad M, Dastugue B, Meiniel A (2001) Effect of SCO-spondin thrombospondin type I repeats (TSR) in comparison to Reissner’s fiber material on the differentiation of the B104 neuroblastoma cell line. Cell Tissue Res 304:361–369
Gobron S, Monnerie H, Meiniel R, Creveaux 1, Lehmann W, Lamalle D, Dastugue B, Meiniel A (1996) SCO-spondin: a new member of the thrombospondin family secreted by the subcommissural organ is a candidate in the modulation of neuronal aggregation. J Cell Sci 109:1053–1061
Gobron S, Creveaux I, Meiniel R, Didier R, Herbet A, Bamdad M, El Bitar F, Dastugue B, Meiniel A (2000) The subcommissural organ/Reissner’s fiber complex: characterization of SCO-spondin, a glycoprotein with potent activity on neurite outgrowth. Glia 32:177–191
Higashijima S, Nose A, Eguchi G, Hotta Y, Okamoto H (1997) Mindin/F-spondin family: novel ECM proteins expressed in the zebrafish embryonic axis. Dev Biol 192:211–227
Ishii T, Satoh E, Nishimura M (2001) Integrin-linked kinase controls neurite outgrowth in N1E-115 neuroblastoma cells. J Biol Chem 42994–43003
Kim JE, Kim SJ, Lee BH, Park RW, Kim KS, Kim IS (2000) Identification of motifs for cell adhesion within the repeated domains of transforming growth factor-beta-induced gene, βig-h3. J Biol Chem 275:30907–30915
Klar A, Baldassare M, Jessel TM (1992) F-spondin gene expressed at light levels in the floor plate encodes a secreted protein that promotes neural cells adhesion and neurite extension. Cell 65:95–110
Lochter A, Taylor KH, Braunewell J, Holm J, Schachner M (1995) Control of neuronal morphology in vitro: interplay between adhesive substrate forces and molecular instruction. J Neurosci Res 42:145–158
Meiniel A (2001) SCO-spondin, a glycoprotein of the subcommissural organ/Reissner’s fiber complex: evidence of a potent activity on neuronal development in primary cell cultures. Microsc Res Tech 52:484–495
Meiniel A, Meiniel R, Didier R, Creveaux 1, Gobron S, Monnerie H, Dastugue B (1996) The subcommissural organ and Reissner’s fibre complex. Prog Histochem Cytochem 30:1–66
Monier-Gavelle F, Duband JL (1997) Cross talk between adhesion molecules: control of N-cadherin activity by intracellular signals elicited by beta1 and beta3 integrins in migrating neural crest cells. J Cell Biol 137:1663–1681
Monnerie H, Boespflug-Tanguy O, Dastugue B, Meiniel A (1996) Soluble material from Reissner’s fibre displays anti-aggregative activity in primary cultures of chick cortical neurons. Dev Brain Res 96:120–129
Monnerie H, Dastugue B, Meiniel A (1998) Effect of synthetic peptides derived from SCO-spondin conserved domains on chick cortical and spinal neurons in cell cultures. Cell Tissue Res 293:407–418
Neugebauer KM, Emmett CJ, Venstrom KA, Relchardt LF (1991) Vitronectin and thrombospondin promote retinal neurite outgrowth: developmental regulation and role of integrins. Neuron 6:345–358
Oksche A, Rodríguez EM, Fernández Llebrez P (1993) The subcommissural organ, an ependymal brain gland. Springer, Berlin Heidelberg New York
O’Shea K, Dixit VM (1998) Unique distribution of the extracellular matrix component thrombospondin in the developing mouse embryo. J Cell Biol 107:2737–2738
O’Shea KS, Rheinheimer JS, Dixit VM (1990) Deposition and role of thrombospondin in the histogenesis of the cerebellar cortex. J Cell Biol 110:1275–1283
O’Shea KS, Liu L-HJ, Dixit VM (1991) Thrombospondin and a 140 kd fragment promote adhesion and neurite outgrowth from embryonic central and peripheral neurons and from PC12 cells. Neuron 7:231–237
Osterhout D, Frazier WA, Higgins D (1992) Thrombospondin promotes process outgrowth in neurons from the peripheral and central nervous systems. Dev Biol 150:256–265
Perris R, Perissinotto D (2000) Role of the extracellular matrix during neural crest cell migration. Mech Dev 95:3–21
Petroski RE, Geller HM (1994) Selective labeling of embryonic neurons cultured on astrocyte monolayers with 5(6)-carboxyfluorescein diacetate (CFDA). J Neurosci Methods 52:23–32
Schmidt C, Künemund V, Wintergerst ES, Schmitz B, Schachner M (1996) CD9 of mouse brain is implicated in neurite outgrowth and cell migration in vitro and is associated with the α6/β1 integrin and the neural adhesion molecule L1. J Neurosci Res 43:12–31
Schubert D, Heinemann S, Carlisle W, Tarikas H, Kimes B, Patrick J, Steinbach JH, Culp W, Brandt BL (1974) Clonal cell lines from the rat central nervous system. Nature 249:224–227
Scott-Drew S, Ffrench-Constant C (1997) Expression and function of thrombospondin-1 in myelinating glial cells of the central nervous system. J Neurosci Res 50:202–214
Shahan TA, Ziaie Z, Pasco S, Fawzi A, Bellon G, Monboisse JC, Kefalides NA (1999) Identification of CD47/integrin-associated protein and alpha (V) beta 3 as two receptors for the alpha 3 (IV) chain of type IV collagen on tumor cells. Cancer Res 59:4584–4590
Simon DI, Wei Y, Rao NK, Xu H, Chen Z, Liu Q, Rosenberg S, Chapman HA (2000) Identification of a urokinase receptor-integrin interaction site. J Biol Chem 275:10288–10234
Testaz S, Delannet M, Duband JL (1999) Adhesion and migration of avian neural crest cells on fibronectin require the cooperating activities of multiple integrins of β1 and β3 families. J Cell Sci 112:4715–4728
Tucker RP, Hagios C, Chiquet-Ehrismann R, Lawler J, Hall RJ, Erickson CA (1999) Thrombospondin-1 and neural crest cell migration. Dev Dyn 214:312–322
Varnum-Finney B, Venstrom K, Muller U, Kypta R, Backus C, Chiquet M, Reichardt LF (1995) The integrin receptor α8β1 mediates interactions of embryonic chick motor and sensory neurons with tenascin C. Neuron 14:1213–1222
Acknowledgments
The authors are grateful to Dr. Jean-Loup Duband for fruitful discussions and for critically reading the manuscript. Thanks are also due to Dr. Chantal Rappatel, Fredérique Lioret and Pascal Pigeon for technical assistance with the flow cytometry.
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Bamdad, M., Volle, D., Dastugue, B. et al. α1β1-Integrin is an essential signal for neurite outgrowth induced by thrombospondin type 1 repeats of SCO-spondin. Cell Tissue Res 315, 15–25 (2004). https://doi.org/10.1007/s00441-003-0793-2
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DOI: https://doi.org/10.1007/s00441-003-0793-2