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Neuropilin-1 mediates collapsin-1/semaphorin III inhibition of endothelial cell motility: functional competition of collapsin-1 and vascular endothelial growth factor-165.

Miao HQ, Soker S, Feiner L, Alonso JL, Raper JA, Klagsbrun M - J. Cell Biol. (1999)

Bottom Line: To determine whether semaphorin/collapsins could interact with NRP1 in nonneuronal cells, the effects of recombinant collapsin-1 on endothelial cells (EC) were examined.Collapsin-1 rapidly disrupted the formation of lamellipodia and induced depolymerization of F-actin in an NRP1-dependent manner.These results suggest that collapsin-1 can inhibit EC motility as well as axon motility, that these inhibitory effects on motility are mediated by NRP1, and that VEGF165 and collapsin-1 compete for NRP1-binding sites.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgical Research, Children's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.

ABSTRACT
Neuropilin-1 (NRP1) is a receptor for two unrelated ligands with disparate activities, vascular endothelial growth factor-165 (VEGF165), an angiogenesis factor, and semaphorin/collapsins, mediators of neuronal guidance. To determine whether semaphorin/collapsins could interact with NRP1 in nonneuronal cells, the effects of recombinant collapsin-1 on endothelial cells (EC) were examined. Collapsin-1 inhibited the motility of porcine aortic EC (PAEC) expressing NRP1 alone; coexpressing KDR and NRP1 (PAEC/KDR/NRP1), but not parental PAEC; or PAEC expressing KDR alone. The motility of PAEC expressing NRP1 was inhibited by 65-75% and this inhibition was abrogated by anti-NRP1 antibody. In contrast, VEGF165 stimulated the motility of PAEC/KDR/NRP1. When VEGF165 and collapsin-1 were added simultaneously to PAEC/KDR/NRP1, dorsal root ganglia (DRG), and COS-7/NRP1 cells, they competed with each other in EC motility, DRG collapse, and NRP1-binding assays, respectively, suggesting that the two ligands have overlapping NRP1 binding sites. Collapsin-1 rapidly disrupted the formation of lamellipodia and induced depolymerization of F-actin in an NRP1-dependent manner. In an in vitro angiogenesis assay, collapsin-1 inhibited the capillary sprouting of EC from rat aortic ring segments. These results suggest that collapsin-1 can inhibit EC motility as well as axon motility, that these inhibitory effects on motility are mediated by NRP1, and that VEGF165 and collapsin-1 compete for NRP1-binding sites.

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Collapsin-1 alters the morphology only of EC expressing NRP1. PAEC (A and E), PAEC/NRP1 (B and F), PAEC/KDR (C and G), and PAEC/KDR/NRP1 (D and H) were incubated for 30 min in the absence (A–D) or presence (E–H) of 300 ng/ml collapsin-1, fixed, and analyzed by DIC optic microscopy as in Fig. 7A–C.
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Figure 8: Collapsin-1 alters the morphology only of EC expressing NRP1. PAEC (A and E), PAEC/NRP1 (B and F), PAEC/KDR (C and G), and PAEC/KDR/NRP1 (D and H) were incubated for 30 min in the absence (A–D) or presence (E–H) of 300 ng/ml collapsin-1, fixed, and analyzed by DIC optic microscopy as in Fig. 7A–C.

Mentions: To determine whether the collapsin-1 inhibition of lamellipodia formation observed for RAEC was mediated by NRP1, parental PAEC and PAEC expressing NRP1, KDR, or both receptors, were treated with collapsin-1 and analyzed by DIC optic microscopy (Fig. 8). Collapsin-1 did not induce morphological changes in parental PAEC (Fig. 8A versus E) or in PAEC/KDR (Fig. 8C versus G). On the other hand, 300 ng/ml collapsin-1 altered the morphology of PAEC/NRP1 (Fig. 8B versus F) and PAEC/KDR/NRP1 (Fig. 8D versus H), by inducing significant retraction of lamellipodia. Taken together, RAEC and PAEC morphology analysis suggest that collapsin-1 disorganizes EC cytoskeleton, retracts lamellipodia, and that these effects are mediated by NRP1.


Neuropilin-1 mediates collapsin-1/semaphorin III inhibition of endothelial cell motility: functional competition of collapsin-1 and vascular endothelial growth factor-165.

Miao HQ, Soker S, Feiner L, Alonso JL, Raper JA, Klagsbrun M - J. Cell Biol. (1999)

Collapsin-1 alters the morphology only of EC expressing NRP1. PAEC (A and E), PAEC/NRP1 (B and F), PAEC/KDR (C and G), and PAEC/KDR/NRP1 (D and H) were incubated for 30 min in the absence (A–D) or presence (E–H) of 300 ng/ml collapsin-1, fixed, and analyzed by DIC optic microscopy as in Fig. 7A–C.
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Related In: Results  -  Collection

Show All Figures
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Figure 8: Collapsin-1 alters the morphology only of EC expressing NRP1. PAEC (A and E), PAEC/NRP1 (B and F), PAEC/KDR (C and G), and PAEC/KDR/NRP1 (D and H) were incubated for 30 min in the absence (A–D) or presence (E–H) of 300 ng/ml collapsin-1, fixed, and analyzed by DIC optic microscopy as in Fig. 7A–C.
Mentions: To determine whether the collapsin-1 inhibition of lamellipodia formation observed for RAEC was mediated by NRP1, parental PAEC and PAEC expressing NRP1, KDR, or both receptors, were treated with collapsin-1 and analyzed by DIC optic microscopy (Fig. 8). Collapsin-1 did not induce morphological changes in parental PAEC (Fig. 8A versus E) or in PAEC/KDR (Fig. 8C versus G). On the other hand, 300 ng/ml collapsin-1 altered the morphology of PAEC/NRP1 (Fig. 8B versus F) and PAEC/KDR/NRP1 (Fig. 8D versus H), by inducing significant retraction of lamellipodia. Taken together, RAEC and PAEC morphology analysis suggest that collapsin-1 disorganizes EC cytoskeleton, retracts lamellipodia, and that these effects are mediated by NRP1.

Bottom Line: To determine whether semaphorin/collapsins could interact with NRP1 in nonneuronal cells, the effects of recombinant collapsin-1 on endothelial cells (EC) were examined.Collapsin-1 rapidly disrupted the formation of lamellipodia and induced depolymerization of F-actin in an NRP1-dependent manner.These results suggest that collapsin-1 can inhibit EC motility as well as axon motility, that these inhibitory effects on motility are mediated by NRP1, and that VEGF165 and collapsin-1 compete for NRP1-binding sites.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgical Research, Children's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.

ABSTRACT
Neuropilin-1 (NRP1) is a receptor for two unrelated ligands with disparate activities, vascular endothelial growth factor-165 (VEGF165), an angiogenesis factor, and semaphorin/collapsins, mediators of neuronal guidance. To determine whether semaphorin/collapsins could interact with NRP1 in nonneuronal cells, the effects of recombinant collapsin-1 on endothelial cells (EC) were examined. Collapsin-1 inhibited the motility of porcine aortic EC (PAEC) expressing NRP1 alone; coexpressing KDR and NRP1 (PAEC/KDR/NRP1), but not parental PAEC; or PAEC expressing KDR alone. The motility of PAEC expressing NRP1 was inhibited by 65-75% and this inhibition was abrogated by anti-NRP1 antibody. In contrast, VEGF165 stimulated the motility of PAEC/KDR/NRP1. When VEGF165 and collapsin-1 were added simultaneously to PAEC/KDR/NRP1, dorsal root ganglia (DRG), and COS-7/NRP1 cells, they competed with each other in EC motility, DRG collapse, and NRP1-binding assays, respectively, suggesting that the two ligands have overlapping NRP1 binding sites. Collapsin-1 rapidly disrupted the formation of lamellipodia and induced depolymerization of F-actin in an NRP1-dependent manner. In an in vitro angiogenesis assay, collapsin-1 inhibited the capillary sprouting of EC from rat aortic ring segments. These results suggest that collapsin-1 can inhibit EC motility as well as axon motility, that these inhibitory effects on motility are mediated by NRP1, and that VEGF165 and collapsin-1 compete for NRP1-binding sites.

Show MeSH
Related in: MedlinePlus