Limits...
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.

Show MeSH

Related in: MedlinePlus

Nrp2-expressing NCCs give rise to neurons and glia of the dorsal root ganglia. (A) Whole mount X-gal staining of E11.5 Nrp2-CreERT2/Kikume X R26R embryos injected with tamoxifen (TM) at E9.5 and E10.0 (arrows). Staining was identified in presumptive skeletal muscle, skin, and DRG. (B-C) Longitudinal sections anterior to the hind limbs counterstained with eosin confirmed X-gal staining within sensory neurons and glia of the DRG, skeletal muscle and skin. (D-I) Transverse sections anterior to the hind limbs counterstained with Tuj1 and TH demonstrated X-gal staining within sensory neurons of the DRG (E-F, dashed circle) but not within sympathetic neurons (G-I, dashed circle). da, dorsal aorta; nt, neural tube. Scale bars = A, 500 μm; D & G, 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4233049&req=5

Fig8: Nrp2-expressing NCCs give rise to neurons and glia of the dorsal root ganglia. (A) Whole mount X-gal staining of E11.5 Nrp2-CreERT2/Kikume X R26R embryos injected with tamoxifen (TM) at E9.5 and E10.0 (arrows). Staining was identified in presumptive skeletal muscle, skin, and DRG. (B-C) Longitudinal sections anterior to the hind limbs counterstained with eosin confirmed X-gal staining within sensory neurons and glia of the DRG, skeletal muscle and skin. (D-I) Transverse sections anterior to the hind limbs counterstained with Tuj1 and TH demonstrated X-gal staining within sensory neurons of the DRG (E-F, dashed circle) but not within sympathetic neurons (G-I, dashed circle). da, dorsal aorta; nt, neural tube. Scale bars = A, 500 μm; D & G, 100 μm.

Mentions: Our expression analysis raised the hypothesis that ventrally migrating trunk NCCs expressing Nrp2 are biased towards neurons and glia of the sensory ganglia. To test this notion, we completed fate-mapping studies with the Nrp2-inducible lineage-tracing mouse model. Tamoxifen was administered to pregnant dams when embryos were E9.5 and E10.0, and these were later collected at E11.5, a stage at which the sensory and sympathetic ganglia have started to condense. Consistent with the staining of Nrp2 in NCCs and paraxial mesoderm at E9.5, X-gal staining identified derivatives of Nrp2-expressing cells within the DRG and skeletal muscle (Figure 8A). Eosin staining of longitudinal sections anterior to the hind limb clearly demonstrated X-gal staining within the DRG (Figure 8B-C). Transverse sections through the same axial level were also counterstained with the pan neuronal marker Tuj1 and the sympathetic specific neuronal marker tyrosine hydroxylase (TH) (Figure 8D). In all cases examined (n = 3), X-gal staining was robustly observed in the DRG (Figure 8D-F). By contrast, we were unable to observe X-gal staining in sympathetic ganglia (Figure 8G-I). This finding demonstrates that Nrp2-expressing NCCs are strongly biased towards neurons and glia of the DRG.Figure 8


Neuropilins define distinct populations of neural crest cells.

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

Nrp2-expressing NCCs give rise to neurons and glia of the dorsal root ganglia. (A) Whole mount X-gal staining of E11.5 Nrp2-CreERT2/Kikume X R26R embryos injected with tamoxifen (TM) at E9.5 and E10.0 (arrows). Staining was identified in presumptive skeletal muscle, skin, and DRG. (B-C) Longitudinal sections anterior to the hind limbs counterstained with eosin confirmed X-gal staining within sensory neurons and glia of the DRG, skeletal muscle and skin. (D-I) Transverse sections anterior to the hind limbs counterstained with Tuj1 and TH demonstrated X-gal staining within sensory neurons of the DRG (E-F, dashed circle) but not within sympathetic neurons (G-I, dashed circle). da, dorsal aorta; nt, neural tube. Scale bars = A, 500 μm; D & G, 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4233049&req=5

Fig8: Nrp2-expressing NCCs give rise to neurons and glia of the dorsal root ganglia. (A) Whole mount X-gal staining of E11.5 Nrp2-CreERT2/Kikume X R26R embryos injected with tamoxifen (TM) at E9.5 and E10.0 (arrows). Staining was identified in presumptive skeletal muscle, skin, and DRG. (B-C) Longitudinal sections anterior to the hind limbs counterstained with eosin confirmed X-gal staining within sensory neurons and glia of the DRG, skeletal muscle and skin. (D-I) Transverse sections anterior to the hind limbs counterstained with Tuj1 and TH demonstrated X-gal staining within sensory neurons of the DRG (E-F, dashed circle) but not within sympathetic neurons (G-I, dashed circle). da, dorsal aorta; nt, neural tube. Scale bars = A, 500 μm; D & G, 100 μm.
Mentions: Our expression analysis raised the hypothesis that ventrally migrating trunk NCCs expressing Nrp2 are biased towards neurons and glia of the sensory ganglia. To test this notion, we completed fate-mapping studies with the Nrp2-inducible lineage-tracing mouse model. Tamoxifen was administered to pregnant dams when embryos were E9.5 and E10.0, and these were later collected at E11.5, a stage at which the sensory and sympathetic ganglia have started to condense. Consistent with the staining of Nrp2 in NCCs and paraxial mesoderm at E9.5, X-gal staining identified derivatives of Nrp2-expressing cells within the DRG and skeletal muscle (Figure 8A). Eosin staining of longitudinal sections anterior to the hind limb clearly demonstrated X-gal staining within the DRG (Figure 8B-C). Transverse sections through the same axial level were also counterstained with the pan neuronal marker Tuj1 and the sympathetic specific neuronal marker tyrosine hydroxylase (TH) (Figure 8D). In all cases examined (n = 3), X-gal staining was robustly observed in the DRG (Figure 8D-F). By contrast, we were unable to observe X-gal staining in sympathetic ganglia (Figure 8G-I). This finding demonstrates that Nrp2-expressing NCCs are strongly biased towards neurons and glia of the DRG.Figure 8

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