Versican V0 and V1 Guide Migratory Neural Crest Cells

Abstract

We previously showed the selective expression of the chondroitin sulfate proteoglycans versican V0 and V1 in barrier tissues that impede the migration of neural crest cells during embryonic trunk development (Landolt, R. M., Vaughan, L., Winterhalter, K. H., and Zimmermann, D. R. (1995) Development 212, 2303-2312). To test for an active involvement of these isoforms in the guidance process, we have now established protocols to isolate intact versican V0 and V1 in quantities sufficient for functional experiments. Using stripe choice assays, we demonstrate that pure preparations of either a mixture of versican V0/V1 or V1 alone strongly inhibit the migration of multipotent Sox10/p75NTR double-positive early neural crest stem cells on fibronectin by interfering with cell-substrate adhesion. We show that this inhibition is largely core glycoprotein-dependent, as the complete removal of the glycosaminoglycan chains has only a minor effect on the inhibitory capacity. Our findings support the notion that versican variants V0 and V1 act, possibly in concert with other inhibitory molecules such as aggrecan and ephrins, in directing the migratory streams of neural crest cells to their appropriate target tissues. We previously showed the selective expression of the chondroitin sulfate proteoglycans versican V0 and V1 in barrier tissues that impede the migration of neural crest cells during embryonic trunk development (Landolt, R. M., Vaughan, L., Winterhalter, K. H., and Zimmermann, D. R. (1995) Development 212, 2303-2312). To test for an active involvement of these isoforms in the guidance process, we have now established protocols to isolate intact versican V0 and V1 in quantities sufficient for functional experiments. Using stripe choice assays, we demonstrate that pure preparations of either a mixture of versican V0/V1 or V1 alone strongly inhibit the migration of multipotent Sox10/p75NTR double-positive early neural crest stem cells on fibronectin by interfering with cell-substrate adhesion. We show that this inhibition is largely core glycoprotein-dependent, as the complete removal of the glycosaminoglycan chains has only a minor effect on the inhibitory capacity. Our findings support the notion that versican variants V0 and V1 act, possibly in concert with other inhibitory molecules such as aggrecan and ephrins, in directing the migratory streams of neural crest cells to their appropriate target tissues. The highly precise and coordinated migration of neural crest cells during early phases of embryonic development is controlled by the differential expression of permissive substrates and non-permissive/inhibitory molecules within the pathways and the bordering tissues, and the set of membrane receptors present on the moving cells (reviewed by Refs. 1Krull C.E. Mech. Dev. 2001; 105: 37-45Crossref PubMed Scopus (64) Google Scholar, 2Bronner-Fraser M. Trends Cell Biol. 1993; 3: 392-397Abstract Full Text PDF PubMed Scopus (60) Google Scholar, 3Erickson C.A. Reedy M.V. Curr. Top. Dev. Biol. 1998; 40: 177-209Crossref PubMed Google Scholar, 4Le Douarin N.M. Mech. Dev. 2004; 121: 1089-1102Crossref PubMed Scopus (101) Google Scholar). The journey of the multipotent neural crest stem and progenitor cells begins in the dorsal neural tube from where they emerge shortly after its closure. In the trunk region the cells are initially guided along a ventral trajectory before a second wave starts to invade the dorsolateral tissue underneath the ectoderm. Whereas the ventrally migrating populations differentiate into neurons and glia of the sensory and the sympathetic nervous system, the laterally progressing cells give rise to the melanocytes of the skin. On their route, neural crest cells pass through highly permissive extracellular matrices, which allow rapid cellular movements. These pathways are flanked by tissues that block neural crest cell immigration and thus provide the directional information. These barrier tissues, previously identified by microsurgical manipulations, include the posterior sclerotomes (5Bronner-Fraser M. Stern C. Dev. Biol. 1991; 143: 213-217Crossref PubMed Scopus (101) Google Scholar), the perinotochordal region (6Pettway Z. Guillory G. Bronner-Fraser M. Dev. Biol. 1990; 142: 335-345Crossref PubMed Scopus (60) Google Scholar), and for a short period also the subectodermal matrix prior to melanocyte precursor invasion (7Erickson C.A. Duong T.D. Tosney K.W. Dev. Biol. 1992; 151: 251-272Crossref PubMed Scopus (131) Google Scholar). Consequently, the streams of neural crest cells, which originally emigrate in an unsegmented fashion from the dorsal neural tube, are on their ventral path canalized into the anterior sclerotome strictly avoiding the posterior somitic halves and the more ventrally localized perinotochordal zone. This particular migration behavior finally leads to the characteristic segmental pattern of the forming sensory and sympathetic ganglia. Since the major migration promoting substrates, fibronectin and laminin (8Rovasio R.A. Delouvee A.D. Yamada K.M. Timple R. Thiery J.P. J. Cell Biol. 1983; 96: 462-473Crossref PubMed Scopus (277) Google Scholar), are uniformly expressed in both halves of the somites (9Rickmann M. Fawcett J.W. Keynes R.J. J. Embryol. Exp. Morph. 1985; 90: 437-455PubMed Google Scholar), the guidance of the migratory neural crest cells appears to depend mainly on inhibitory cues. Several extracellular matrix and cell surface components match the candidate profile for a migration blocking function as they are selectively expressed in non-permissive tissues. Molecules consistently absent from the pathways, but highly expressed within the barriers, include chondroitin 6-sulfate proteoglycans, peanut agglutinin (PNA) 4The abbreviations used are: PNA, peanut agglutinin; eNCSCs, early neural crest stem cells; GdnHCl, guanidine hydrochloride; PBS, phosphate-buffered saline; GAG, glycosaminoglycan; ADAMTS, a disintegrin and metalloproteinase with thrombospondinlike motifs. -binding glycoproteins (10Oakley R.A. Lasky C.J. Erickson C.A. Tosney K.W. Development. 1994; 120: 103-114Crossref PubMed Google Scholar, 11Oakley R.A. Tosney K.W. Dev. Biol. 1991; 147: 187-206Crossref PubMed Scopus (240) Google Scholar), F-spondin (12Debby-Brafman A. Burstyn-Cohen T. Klar A. Kalcheim C. Neuron. 1999; 22: 475-488Abstract Full Text Full Text PDF PubMed Scopus (97) Google Scholar), semaphorin3A (13Eickholt B.J. Mackenzie S.L. Graham A. Walsh F.S. Doherty P. Development. 1999; 126: 2181-2189Crossref PubMed Google Scholar), T-cadherin (14Ranscht B. Bronner-Fraser M. Development. 1991; 111: 15-22Crossref PubMed Google Scholar), collagen IX (15Ring C. Hassell J. Halfter W. Dev. Biol. 1996; 180: 41-53Crossref PubMed Scopus (64) Google Scholar), and, except for the dorsolateral path, ephrins (16Krull C.E. Lansford R. Gale N.W. Collazo A. Marcelle C. Yancopoulos G.D. Fraser S.E. Bronner-Fraser M. Curr. Biol. 1997; 7: 571-580Abstract Full Text Full Text PDF PubMed Scopus (337) Google Scholar, 17Santiago A. Erickson C.A. Development. 2002; 129: 3621-3632Crossref PubMed Google Scholar, 18Wang H.U. Anderson D.J. Neuron. 1997; 18: 383-396Abstract Full Text Full Text PDF PubMed Scopus (419) Google Scholar). For some of these molecules, like semaphorin3A (13Eickholt B.J. Mackenzie S.L. Graham A. Walsh F.S. Doherty P. Development. 1999; 126: 2181-2189Crossref PubMed Google Scholar) and ephrins (ephrin-B1 in avian (16Krull C.E. Lansford R. Gale N.W. Collazo A. Marcelle C. Yancopoulos G.D. Fraser S.E. Bronner-Fraser M. Curr. Biol. 1997; 7: 571-580Abstract Full Text Full Text PDF PubMed Scopus (337) Google Scholar) and ephrin-B2 in mammalian embryos (18Wang H.U. Anderson D.J. Neuron. 1997; 18: 383-396Abstract Full Text Full Text PDF PubMed Scopus (419) Google Scholar)), inhibitory activities on neural crest cell migration have been demonstrated in vitro using neural tube or whole trunk explant culture systems. Unexpectedly, however, neither the gene inactivation of semaphorin3A (19Kawasaki T. Bekku Y. Suto F. Kitsukawa T. Taniguchi M. Nagatsu I. Nagatsu T. Itoh K. Yagi T. Fujisawa H. Development. 2002; 129: 671-680Crossref PubMed Google Scholar) nor of ephrin-B2 (20Wang H.U. Chen Z.F. Anderson D.J. Cell. 1998; 93: 741-753Abstract Full Text Full Text PDF PubMed Scopus (1382) Google Scholar) and the corresponding Eph receptors on neural crest cells (21Henkemeyer M. Orioli D. Henderson J.T. Saxton T.M. Roder J. Pawson T. Klein R. Cell. 1996; 86: 35-46Abstract Full Text Full Text PDF PubMed Scopus (465) Google Scholar, 22Orioli D. Henkemeyer M. Lemke G. Klein R. Pawson T. EMBO J. 1996; 15: 6035-6049Crossref PubMed Scopus Google Scholar) in the in migration through the These that a of inhibitory and some are to trunk neural crest cells in C.E. Mech. Dev. 2001; 105: 37-45Crossref PubMed Scopus (64) Google Scholar, 17Santiago A. Erickson C.A. Development. 2002; 129: 3621-3632Crossref PubMed Google Scholar, P. 1998; PubMed Scopus Google Scholar, K.W. Dev. 2004; PubMed Scopus Google Scholar). for a with the cell surface of neural crest are extracellular matrix components to the chondroitin sulfate proteoglycans and glycoproteins (10Oakley R.A. Lasky C.J. Erickson C.A. Tosney K.W. Development. 1994; 120: 103-114Crossref PubMed Google Scholar, 11Oakley R.A. Tosney K.W. Dev. Biol. 1991; 147: 187-206Crossref PubMed Scopus (240) Google Scholar, B. Dev. Biol. 1990; PubMed Scopus Google Scholar, R. D. T. C. Bronner-Fraser M. Development. 1991; 111: PubMed Google Scholar). the chondroitin sulfate proteoglycans versican EMBO J. PubMed Scopus Google Scholar) and aggrecan K. M. Hassell Yamada Y. J. Biol. Full Text PDF PubMed Google Scholar) to in the proteoglycans are of the forming through with and C.E. PubMed Scopus Google Scholar). during as the of their expression leads in to early or C. H. A. D. R. Dev. Biol. 1998; PubMed Scopus Google Scholar, H. K. D. C.A. Yamada Y. 1994; 7: PubMed Scopus Google Scholar). isoforms of versican to as a of of the glycosaminoglycan glycosaminoglycan and J. Biol. 1994; Full Text PDF PubMed Google Scholar, M. T. M. K. K. J. Biol. Full Text Full Text PDF PubMed Scopus Google Scholar). The variants of V0 and are highly expressed during and are with tissues a extracellular matrix M. T. K. Embryol. 1993; PubMed Scopus Google Scholar, Development. 121: PubMed Google Scholar). a nervous is in during P. P. A. M. Y. 1998; PubMed Scopus Google Scholar). is a of and to in the of and impede in the nervous M. C.E. J. Cell PubMed Google Scholar). We have previously that the expression of versican V0 and V1 is during neural crest cell migration with the of barrier tissues in the trunk of embryos Development. 121: PubMed Google Scholar). V0 and V1 are selectively in the posterior of the present within the dorsolateral subectodermal tissues and to a expressed in the perinotochordal tissue where aggrecan appears to the M. H. A. J. Dev. Biol. PubMed Scopus Google Scholar, Z. M. Bronner-Fraser M. Exp. Cell 1996; PubMed Scopus Google Scholar). This of versican V0 and V1 with barrier tissues to isolate the intact of these versican variants and to their guidance function in neural crest cell migration during early embryonic trunk of the or of and from previously in J. Biol. 1994; Full Text PDF PubMed Google Scholar, M. C.E. J. Cell PubMed Google Scholar, M. J. Biol. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar, M. J. Cell Biol. 1994; PubMed Scopus Google Scholar). intact chondroitin sulfate chains and for chondroitin 6-sulfate after from and from and a for C. M. Development. 2001; PubMed Google Scholar) a of of For of fibronectin the of and a mixture of V0 and V1 from and the culture of the cell Our previously for the of versican for this M. J. Biol. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar). and in the of the tissues from the in of guanidine and and through the by for The The during by for sulfate to the to to for the sulfate to an the and in by of the This on and into a The with and with a to of for the of by using or M. J. Biol. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar). on the and to which previously been to the of A. PubMed Scopus Google Scholar) by from to used for versican to The and with a of to with GdnHCl, with the and to a of using For the V1 from an on to For this the with of the before of the The a of second for the of versican V1 on a PBS, and For the of a mixture of V0 and V1 from cell the proteoglycans from by sulfate to by with and as the versican used for the To core to of the versican preparations with of in to with a and on and with either or with For on by for The with in for by with the in the blocking or in to for The with of of and from neural of embryonic to the of M. H. J. D.J. J. Cell Biol. PubMed Scopus Google Scholar, M. Cell Scholar). embryos of the with the trunk region of the to the posterior from the of the with the of and The and to a in and The and for This using a the neural to of other tissues and to to the The on substrates in with of the in a with and M. H. J. D.J. J. Cell Biol. PubMed Scopus Google Scholar). from the trunk of embryos as by C. R. Biol. 1999; 129: Google Scholar). choice as previously M. C.E. J. Cell PubMed Google Scholar) from J. B. F. Exp. 1990; PubMed Scopus Google in of test and substrates for migratory The of the test of versican isoforms to to the with fibronectin To major in the of fibronectin of versican to the test of of and using the region of For to neural to the stripe with culture and for to in a the of the the to the the with and For this the in a which for to with a mixture and to to of to of the neural tube for and with in for either by or for embryonic to emigrate the stripe pattern from the surface of Cell the assays, from embryos as M. R. 2004; PubMed Scopus Google Scholar) and after in culture on with fibronectin alone versican or a mixture of versican and fibronectin and The and of the by the cells after or of The of cells that after on fibronectin alone as of and by of with and with a C. M. Development. 2001; PubMed Google For this cells and with and for they with the for and for with and the in some with the of versican and with the For of tissue embryos and in of the tissue in and by a of in of the in in a for The in for and in and in for and in blocking from a The with the and the for versican to in in in PBS, the with and in with for finally in and in with an with a using the and in the corresponding in and with of the of and of the isoforms of versican for functional has been as they except for versican expressed in embryonic tissues only quantities of the intact proteoglycans K. Y. K. T. M. M. J. Biol. Full Text PDF PubMed Google Scholar). In tissues such as of B. J. J. 1996; PubMed Scopus Google Scholar). this however, versican is largely present in from a other matrix J. C. J.W. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). We for of intact in tissues of and in of cells in previously to versican from M. J. Biol. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar), we to isolate a mixture of versican V0/V1 from the culture of the cell versican V1 with of V0 from and for versican from the isoforms intact their characteristic core on only after removal of the chondroitin sulfate chains and migrating in the of the on of the and preparations with an with the which this for some components by the with other in and The of these preparations of versican V0/V1 of culture of versican V1 of and of versican of to the V0/V1 and V1 the of in sufficient of intact we now their effect on the migration behavior of in These in vitro the expression pattern of versican V0 and V1 within the posterior sclerotome during the active neural crest cell migration period in Development. 121: PubMed Google Scholar) and in embryos Consequently, neural from embryos used as the stem cell for the in vitro experiments. The on with of the extracellular matrix fibronectin and the versican test to in the from the neural tube as and the This migration in assays, in which the with and fibronectin in a stripe pattern The however, with of fibronectin with with a mixture of versican V0 and V1 fibronectin In these the of the migratory neural crest cells into streams shortly after the neural tube only on the This selective from the behavior of embryonic trunk which a In a stripe choice the a versican the migration of embryonic and that embryonic which show in to in an stripe choice migration In to the early embryonic neural crest stem cells versican V0/V1 substrates of versican as as and the migration promoting effect of fibronectin also present in these Whereas the inhibition complete this versican V0/V1 of the cells and and and with intact versican V1 from in this In to the effect on migration the with versican substrates to have on cellular with the and characteristic of an early neural crest stem cell C. M. Development. 2001; PubMed Google Scholar, M. R. 2004; PubMed Scopus Google Scholar), the versican a along the migration path the after of the the inhibitory function of versican V0/V1 from the or from the core we preparations with and in stripe choice to these the removal of the chondroitin sulfate chains been by and with the intact chondroitin sulfate and the of chondroitin 6-sulfate to the core region after and intact versican V0/V1 in the a but in a after we that the more The stripe choice demonstrated that the versican core glycoproteins to the migration of to the with the intact proteoglycans, the effect to the of the versican core some of the inhibitory of cells migrating of the neural tube showed only a stripe of and the cells the stripe pattern migrating on with fibronectin This set of showed that the to inhibit migration within the versican core The chondroitin sulfate chains however, to this inhibitory the to after the removal to a effect as with the intact with the of to intact versican with cell and populations on culture with versican V0/V1 alone fibronectin alone and a mixture of versican and fibronectin and These demonstrated that in to the of cells to to the surface with versican V0/V1 on pure versican V0/V1 cell after the cell in the these in vitro that inhibit neural crest cell migration by the of neural crest cells to extracellular matrix substrates such as The findings of present and Development. 121: PubMed Google Scholar) provide for a functional involvement of V0 and V1 in the inhibition and guidance of neural crest cell migration during embryonic trunk These are in the coordinated expression of versican V0 and V1 in barrier tissues of and embryos the invasion of neural crest stem and progenitor cells; the of from intact versican V0 and V1 in stripe choice the mainly core inhibitory function of interfering with migration after complete removal of chondroitin sulfate and the of in the of to their for functional has been the of for the of the intact This has in the been by the that the versican variants V0 and V1 are expressed during early phases of embryonic development M. T. K. Embryol. 1993; PubMed Scopus Google Scholar, Development. 121: PubMed Google Scholar), where they are to a highly controlled by the of and matrix J. C. J.W. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar, J. A. G. J. 2004; PubMed Scopus Google Scholar). Consequently, the of intact versican are embryonic tissues are used as a of core from the of embryos K. Y. K. T. M. M. J. Biol. Full Text PDF PubMed Google the expression of the V0 and V1 isoforms is and the core in the are largely as demonstrated by of tissue and by of after with J. C. J.W. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar, J. A. G. J. 2004; PubMed Scopus Google Scholar). this V0 and V1 in tissues such as B. J. J. 1996; PubMed Scopus Google Scholar, C. Biol. 1994; PubMed Scopus Google Scholar) and M. J. Cell Biol. 1994; PubMed Scopus Google Scholar) and by Whereas these to the of extracellular matrices, appears that the of V0 and V1 is for the of their in cell and migration during embryonic the of versican V0 and V1 we have used tissues from and from cell to sufficient quantities of highly and intact proteoglycans for functional In the of in these of versican migrating on and on from the intact core their with versican they are from versican V0 by within the This is from the that they versican with and from tissue expression J. Biol. 1994; Full Text PDF PubMed Google Scholar) and we have that versican is expressed of the nervous as has been in a D. P. I. R. A. Z. Bronner-Fraser M. T. K. M. J. R. Development. PubMed Google Scholar). Whereas the intact versican V1 with only of versican a of these isoforms from the versican isoforms after strongly with the used in (10Oakley R.A. Lasky C.J. Erickson C.A. Tosney K.W. Development. 1994; 120: 103-114Crossref PubMed Google Scholar, 11Oakley R.A. Tosney K.W. Dev. Biol. 1991; 147: 187-206Crossref PubMed Scopus (240) Google Scholar, B. Dev. Biol. 1990; PubMed Scopus Google Scholar, R. D. T. C. Bronner-Fraser M. Development. 1991; 111: PubMed Google Scholar). isoforms of versican also the peanut and D. R. Zimmermann, appears that of the and are to the core of versican V0 and V1 in tissues forming to neural crest cell migration and The of versican V0 and V1 and the previously inhibitory glycoproteins from the somites Stern Keynes R.J. Neuron. 1990; Full Text PDF PubMed Scopus Google Scholar) is cell of embryonic of and the V0 and we have of functional with preparations a mixture of these versican V1 alone to in In stripe choice assays, fibronectin to the versican This we have that as non-permissive substrates for migratory neural crest stem cells, but function as active which the of fibronectin is in the embryonic trunk present in both pathways and barrier tissues (9Rickmann M. Fawcett J.W. Keynes R.J. J. Embryol. Exp. Morph. 1985; 90: 437-455PubMed Google Scholar), the stripe choice are the in In to other inhibitory molecules that have previously been in a (13Eickholt B.J. Mackenzie S.L. Graham A. Walsh F.S. Doherty P. Development. 1999; 126: 2181-2189Crossref PubMed Google Scholar, C. Hassell J. Halfter W. Dev. Biol. 1996; 180: 41-53Crossref PubMed Scopus (64) Google Scholar, 18Wang H.U. Anderson D.J. Neuron. 1997; 18: 383-396Abstract Full Text Full Text PDF PubMed Scopus (419) Google Scholar), only versican to to versican V0 and V1 the of neural crest cell The of functional are in with that the of chondroitin sulfate in embryos leads to an migration behavior of neural crest cells through the posterior sclerotomes Y. T. M. T. K. Dev. 1999; PubMed Scopus Google Scholar), which of chondroitin sulfate proteoglycans and in particular the versican isoforms V0 and V1 Development. 121: PubMed Google Scholar). by in which the of neural crest cell migration and the of target tissue with the expression of versican in the migratory pathways of these D.J. P. Mech. Dev. 1997; PubMed Scopus Google Scholar). a the inhibitory of on neural crest cell migration has been a for the aggrecan D. P. I. R. A. Z. Bronner-Fraser M. T. K. M. J. R. Development. PubMed Google Scholar). In this D. P. I. R. A. Z. Bronner-Fraser M. T. K. M. J. R. Development. PubMed Google Scholar) a function of has been demonstrate the effect of the intact V0 and This by the that versican preparations from and used in this the of with of the to the inhibitory of these preparations have been the through the of moving neural crest cells with versican is into the inhibition of cellular migration is either a on the surface of neural crest cells or by interfering with the migration promoting substrates and their receptors M. J. 1994; PubMed Scopus Google Scholar, M. J. J. Cell 1999; PubMed Google Scholar). The is by the that the complete removal of the chondroitin sulfate chains leads only to a but to the of the inhibitory This effect by a of the core in the of the highly glycosaminoglycan the of the versican and possibly the of a the cell surface and the permissive this appears that the of inhibition is in controlled by the expression of in core and of chondroitin sulfate chains to the that has been identified as the however, we have that the nervous versican M. C.E. J. Cell PubMed Google Scholar). R. B. T. G. A. C.E. Cell 2004; PubMed Scopus Google Scholar) demonstrate that this inhibition is by a of the of and This has that the with versican and in Consequently, the of an versican has been that this finally to a also for the inhibition of neural crest cell migration by versican V0 and cellular the of is in with of migratory cells, the of the of membrane the 7: PubMed Scopus Google Scholar). receptors are of these C. D. F. T. R. J. 2002; PubMed Scopus Google Scholar). For the of and its the migration of by the of the to its R.A. K. Curr. Cell Biol. 2001; PubMed Scopus Google Scholar). the is for its in the of neural crest cells C.E. G. D. Bronner-Fraser M. Dev. Biol. 1998; PubMed Scopus Google Scholar, Dev. 2001; PubMed Scopus Google Scholar). a with versican V0 and V1 also the migration of neural crest cells during embryonic The of by versican V0 and V1 sufficient to neural crest cell invasion of barrier tissues in as some neural crest cells from a F. Development. PubMed Scopus Google Scholar) to a migration pattern (reviewed in P. K. 3: PubMed Scopus Google Scholar). For a cells, such an within the of the sclerotome has been in trunk after the the on migratory neural crest cells and on the surface of posterior cells by of C.E. Mech. Dev. 2001; 105: 37-45Crossref PubMed Scopus (64) Google Scholar, C.E. Lansford R. Gale N.W. Collazo A. Marcelle C. Yancopoulos G.D. Fraser S.E. Bronner-Fraser M. Curr. Biol. 1997; 7: 571-580Abstract Full Text Full Text PDF PubMed Scopus (337) Google Scholar). inactivation of the corresponding or Eph alone has to an in to trunk neural crest cell migration (20Wang H.U. Chen Z.F. Anderson D.J. Cell. 1998; 93: 741-753Abstract Full Text Full Text PDF PubMed Scopus (1382) Google Scholar). This that the cell ephrins and the extracellular V0 and V1 the migration of neural crest stem and progenitor cells through the possibly have to on of to ephrins, and the other migration in in their to the highly precise migration of the neural crest cell in a We for the for the of the tissue for the and and for