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Neuropilins define distinct populations of neural crest cells.

Lumb R, Wiszniak S, Kabbara S, Scherer M, Harvey N, Schwarz Q - Neural Dev (2014)

Bottom Line: Taken together, our results demonstrate that Nrp1 and Nrp2 are expressed in different populations of NCCs, and that Nrp2-expressing NCCs are strongly biased towards a sensory fate.In the trunk, Nrp2-expressing NCCs specifically give rise to sensory ganglia, whereas Nrp1-expressing NCCs likely give rise to both sensory and sympathetic ganglia.Our findings therefore suggest that neuropilins play an essential role in coordinating NCC migration with fate specification.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cancer Biology, University of South Australia and SA Pathology, Frome Road, Adelaide 5000, Australia. quenten.schwarz@health.sa.gov.au.

ABSTRACT

Background: Neural crest cells (NCCs) are a transient embryonic cell type that give rise to a wide spectrum of derivatives, including neurons and glia of the sensory and autonomic nervous system, melanocytes and connective tissues in the head. Lineage-tracing and functional studies have shown that trunk NCCs migrate along two distinct paths that correlate with different developmental fates. Thus, NCCs migrating ventrally through the anterior somite form sympathetic and sensory ganglia, whereas NCCs migrating dorsolaterally form melanocytes. Although the mechanisms promoting migration along the dorsolateral path are well defined, the molecules providing positional identity to sympathetic and sensory-fated NCCs that migrate along the same ventral path are ill defined. Neuropilins (Nrp1 and Nrp2) are transmembrane glycoproteins that are essential for NCC migration. Nrp1 and Nrp2 knockout mice have disparate phenotypes, suggesting that these receptors may play a role in sorting NCCs biased towards sensory and sympathetic fates to appropriate locations.

Results: Here we have combined in situ hybridisation, immunohistochemistry and lineage-tracing analyses to demonstrate that neuropilins are expressed in a non-overlapping pattern within NCCs. Whereas Nrp1 is expressed in NCCs emigrating from hindbrain rhombomere 4 (r4) and within trunk NCCs giving rise to sympathetic and sensory ganglia, Nrp2 is preferentially expressed in NCCs emigrating from r2 and in trunk NCCs giving rise to sensory ganglia. By generating a tamoxifen-inducible lineage-tracing system, we further demonstrate that Nrp2-expressing NCCs specifically populate sensory ganglia including the trigeminal ganglia (V) in the head and the dorsal root ganglia in the trunk.

Conclusions: Taken together, our results demonstrate that Nrp1 and Nrp2 are expressed in different populations of NCCs, and that Nrp2-expressing NCCs are strongly biased towards a sensory fate. In the trunk, Nrp2-expressing NCCs specifically give rise to sensory ganglia, whereas Nrp1-expressing NCCs likely give rise to both sensory and sympathetic ganglia. Our findings therefore suggest that neuropilins play an essential role in coordinating NCC migration with fate specification.

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Nrp1 and Nrp2 are expressed in distinct populations of trunk NCCs. (A-E) Lateral view of whole mount immunofluorescence staining of Nrp1, Nrp2 and trunk NCCs at E9.5. (C) GFP labelled all NCCs migrating in the anterior half of the somite. (D) Nrp1 was expressed in many GFP-positive NCCs and intersomitic vessels. (E) Nrp2 was expressed in the anterior half of the somite and within NCCs. Notably, Nrp1 had high expression in NCCs that had either low or negative Nrp2 expression (white arrow). (F) Transverse sections through the anterior half of the somite over the forelimb region showed that neuropilins were expressed within different populations of NCCs. NCCs migrating through the somite in and around the anlage of the dorsal root ganglia (Fi-i′′′) consisted of cells with high Nrp2 and no Nrp1 expression (inset) and conversely, with high levels of Nrp1 and no expression of Nrp2 (white arrow). (Fii-ii′′′) NCCs migrating towards the dorsal aorta (inset) or condensing in the region of the sympathetic ganglia (white arrow) preferentially expressed Nrp1. av, anterior cardinal vein; bv, blood vessel; da, dorsal aorta. Scale bars = 100 μm.
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Fig4: Nrp1 and Nrp2 are expressed in distinct populations of trunk NCCs. (A-E) Lateral view of whole mount immunofluorescence staining of Nrp1, Nrp2 and trunk NCCs at E9.5. (C) GFP labelled all NCCs migrating in the anterior half of the somite. (D) Nrp1 was expressed in many GFP-positive NCCs and intersomitic vessels. (E) Nrp2 was expressed in the anterior half of the somite and within NCCs. Notably, Nrp1 had high expression in NCCs that had either low or negative Nrp2 expression (white arrow). (F) Transverse sections through the anterior half of the somite over the forelimb region showed that neuropilins were expressed within different populations of NCCs. NCCs migrating through the somite in and around the anlage of the dorsal root ganglia (Fi-i′′′) consisted of cells with high Nrp2 and no Nrp1 expression (inset) and conversely, with high levels of Nrp1 and no expression of Nrp2 (white arrow). (Fii-ii′′′) NCCs migrating towards the dorsal aorta (inset) or condensing in the region of the sympathetic ganglia (white arrow) preferentially expressed Nrp1. av, anterior cardinal vein; bv, blood vessel; da, dorsal aorta. Scale bars = 100 μm.

Mentions: To investigate whether neuropilins are co-expressed or differentially expressed within distinct populations of ventrally migrating NCCs, we performed immunofluorescence analysis on E9.5 Wnt1Cre; Z/EG embryos. Whole mount staining for GFP, Nrp1 and Nrp2 identified mixed populations of NCCs expressing the neuropilin receptors (Figure 4A-E). Consistent with our in situ hybridisation analysis results, Nrp1 was identified in NCCs in the anterior half of the somite, the intersomitic blood vessels, and the dorsal aorta (Figure 4D). Nrp2 was also expressed in NCCs in the anterior half of the somite in addition to the intersomitic blood vessels. Notably, there was limited overlap of the Nrp1 and Nrp2 expression domains, and NCCs expressing high levels of Nrp1 with low or absent Nrp2 could be identified in the dorsal regions of the somite (Figure 4A-E arrow).Figure 4


Neuropilins define distinct populations of neural crest cells.

Lumb R, Wiszniak S, Kabbara S, Scherer M, Harvey N, Schwarz Q - Neural Dev (2014)

Nrp1 and Nrp2 are expressed in distinct populations of trunk NCCs. (A-E) Lateral view of whole mount immunofluorescence staining of Nrp1, Nrp2 and trunk NCCs at E9.5. (C) GFP labelled all NCCs migrating in the anterior half of the somite. (D) Nrp1 was expressed in many GFP-positive NCCs and intersomitic vessels. (E) Nrp2 was expressed in the anterior half of the somite and within NCCs. Notably, Nrp1 had high expression in NCCs that had either low or negative Nrp2 expression (white arrow). (F) Transverse sections through the anterior half of the somite over the forelimb region showed that neuropilins were expressed within different populations of NCCs. NCCs migrating through the somite in and around the anlage of the dorsal root ganglia (Fi-i′′′) consisted of cells with high Nrp2 and no Nrp1 expression (inset) and conversely, with high levels of Nrp1 and no expression of Nrp2 (white arrow). (Fii-ii′′′) NCCs migrating towards the dorsal aorta (inset) or condensing in the region of the sympathetic ganglia (white arrow) preferentially expressed Nrp1. av, anterior cardinal vein; bv, blood vessel; da, dorsal aorta. Scale bars = 100 μm.
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Related In: Results  -  Collection

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Fig4: Nrp1 and Nrp2 are expressed in distinct populations of trunk NCCs. (A-E) Lateral view of whole mount immunofluorescence staining of Nrp1, Nrp2 and trunk NCCs at E9.5. (C) GFP labelled all NCCs migrating in the anterior half of the somite. (D) Nrp1 was expressed in many GFP-positive NCCs and intersomitic vessels. (E) Nrp2 was expressed in the anterior half of the somite and within NCCs. Notably, Nrp1 had high expression in NCCs that had either low or negative Nrp2 expression (white arrow). (F) Transverse sections through the anterior half of the somite over the forelimb region showed that neuropilins were expressed within different populations of NCCs. NCCs migrating through the somite in and around the anlage of the dorsal root ganglia (Fi-i′′′) consisted of cells with high Nrp2 and no Nrp1 expression (inset) and conversely, with high levels of Nrp1 and no expression of Nrp2 (white arrow). (Fii-ii′′′) NCCs migrating towards the dorsal aorta (inset) or condensing in the region of the sympathetic ganglia (white arrow) preferentially expressed Nrp1. av, anterior cardinal vein; bv, blood vessel; da, dorsal aorta. Scale bars = 100 μm.
Mentions: To investigate whether neuropilins are co-expressed or differentially expressed within distinct populations of ventrally migrating NCCs, we performed immunofluorescence analysis on E9.5 Wnt1Cre; Z/EG embryos. Whole mount staining for GFP, Nrp1 and Nrp2 identified mixed populations of NCCs expressing the neuropilin receptors (Figure 4A-E). Consistent with our in situ hybridisation analysis results, Nrp1 was identified in NCCs in the anterior half of the somite, the intersomitic blood vessels, and the dorsal aorta (Figure 4D). Nrp2 was also expressed in NCCs in the anterior half of the somite in addition to the intersomitic blood vessels. Notably, there was limited overlap of the Nrp1 and Nrp2 expression domains, and NCCs expressing high levels of Nrp1 with low or absent Nrp2 could be identified in the dorsal regions of the somite (Figure 4A-E arrow).Figure 4

Bottom Line: Taken together, our results demonstrate that Nrp1 and Nrp2 are expressed in different populations of NCCs, and that Nrp2-expressing NCCs are strongly biased towards a sensory fate.In the trunk, Nrp2-expressing NCCs specifically give rise to sensory ganglia, whereas Nrp1-expressing NCCs likely give rise to both sensory and sympathetic ganglia.Our findings therefore suggest that neuropilins play an essential role in coordinating NCC migration with fate specification.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cancer Biology, University of South Australia and SA Pathology, Frome Road, Adelaide 5000, Australia. quenten.schwarz@health.sa.gov.au.

ABSTRACT

Background: Neural crest cells (NCCs) are a transient embryonic cell type that give rise to a wide spectrum of derivatives, including neurons and glia of the sensory and autonomic nervous system, melanocytes and connective tissues in the head. Lineage-tracing and functional studies have shown that trunk NCCs migrate along two distinct paths that correlate with different developmental fates. Thus, NCCs migrating ventrally through the anterior somite form sympathetic and sensory ganglia, whereas NCCs migrating dorsolaterally form melanocytes. Although the mechanisms promoting migration along the dorsolateral path are well defined, the molecules providing positional identity to sympathetic and sensory-fated NCCs that migrate along the same ventral path are ill defined. Neuropilins (Nrp1 and Nrp2) are transmembrane glycoproteins that are essential for NCC migration. Nrp1 and Nrp2 knockout mice have disparate phenotypes, suggesting that these receptors may play a role in sorting NCCs biased towards sensory and sympathetic fates to appropriate locations.

Results: Here we have combined in situ hybridisation, immunohistochemistry and lineage-tracing analyses to demonstrate that neuropilins are expressed in a non-overlapping pattern within NCCs. Whereas Nrp1 is expressed in NCCs emigrating from hindbrain rhombomere 4 (r4) and within trunk NCCs giving rise to sympathetic and sensory ganglia, Nrp2 is preferentially expressed in NCCs emigrating from r2 and in trunk NCCs giving rise to sensory ganglia. By generating a tamoxifen-inducible lineage-tracing system, we further demonstrate that Nrp2-expressing NCCs specifically populate sensory ganglia including the trigeminal ganglia (V) in the head and the dorsal root ganglia in the trunk.

Conclusions: Taken together, our results demonstrate that Nrp1 and Nrp2 are expressed in different populations of NCCs, and that Nrp2-expressing NCCs are strongly biased towards a sensory fate. In the trunk, Nrp2-expressing NCCs specifically give rise to sensory ganglia, whereas Nrp1-expressing NCCs likely give rise to both sensory and sympathetic ganglia. Our findings therefore suggest that neuropilins play an essential role in coordinating NCC migration with fate specification.

Show MeSH
Related in: MedlinePlus