Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2005 May 17;102(20):7192-7.
doi: 10.1073/pnas.0502581102. Epub 2005 May 9.

Late-emigrating neural crest cells in the roof plate are restricted to a sensory fate by GDF7

Liching Lo  1 , Emma L Dormand, David J Anderson
Affiliations
Comparative Study

Late-emigrating neural crest cells in the roof plate are restricted to a sensory fate by GDF7

Liching Lo et al. Proc Natl Acad Sci U S A. .

Abstract

Lineage-tracing experiments have shown that some premigratory neural crest cells generate both sensory (S) and autonomic (A) derivatives, whereas others generate only S derivatives. Whether this lineage heterogeneity reflects random variation in a homogeneous population or an early sensory specification of some premigratory crest cells has not been clear. Using Cre recombinase-based fate mapping, we show that GDF7, which is exclusively expressed in the roof plate, marks neural crest cells with a 10-fold higher bias to the sensory lineage than those marked (at the same stage of development) by an inducible Wnt1-Cre, which is expressed more broadly in the dorsal neural tube. In vitro, GDF7 has potent sensory neuron-inducing activity. These data suggest that some premigratory crest cells are deterministically restricted to the S lineage and implicate GDF7 itself in this restriction process.

PubMed Disclaimer

Figures

Fig. 1.
Fig. 1.
GDF7 marks a premigratory neural crest subpopulation that is strongly biased to a S fate. (A-F) Double-label immunofluorescence of cross sections through an E9 GDF7Cre; Rosa26STOPlacZ embryo stained with antibodies to β-galactosidase (A and D) or Sox10 (B and E). (A-C) Note that β-gal+ cells are restricted to the roof-plate (A). (G and I) Section of E12.5 GDF7-Cre; Rosa26STOPlacZ embryo stained with X-gal shows β-gal+ cells in the dorsal root ganglia (G) but not in the sympathetic ganglia (I). (H and J) Section through a control E12.5 Wnt1-CreER; Rosa26STOPlacZ embryo injected with 4-OH Tamoxifen at E9.5 reveals β-gal+ cells in both the dorsal root ganglia (H) and sympathetic ganglia (J, arrows). (K and L) Double-labeling for β-gal and TH, a marker of sympathetic neurons (arrows). See Table 1 for quantification. (M) Whole-mount staining of a Gdf7-Cre; Rosa26STOPlacZ embryo at E9.5 reveals that Gdf7-expressing cells extend along the dorsal neural tube to caudal-most regions of the embryo (arrow and Inset), where sympathetic as well as S neurons are being generated at this stage.
Fig. 2.
Fig. 2.
GDF7-Cre-derived neural crest cells generate both S neurons and glia. Analysis of GDF7-Cre; Rosa26STOPlacZ embryos (A-A″ and C-C″) or Wnt1CreER; Rosa26STOPlacZ embryos (B-B″ and D-D″) at E12.5. Double-labeling with antibodies to Brn3a (A, A″, B, and B″, green) and β-gal (A, A′, B, and B′, red) reveals S neurons (A and B, yellow). Double-labeling with antibodies to Sox10 (C″ and D″, green) and β-gal (C′ and D′, red) and shows that GDF7-Cre- and Wnt1-CreER-derived neural crest cells also generate glia (C and D, yellow).
Fig. 3.
Fig. 3.
GDF7 selectively promotes S differentiation in dNT cultures. E10 rat dNT cultures were grown for 3 days in the indicated concentrations of growth factors, fixed, and double-labeled with anti-Phox2b (A-D) and anti-Brn3a (A′-D′). See Fig. 4 for quantification. (E-E′) GDF7 induces Brn3a+ peripheral S neurons and not Brn3a+ dorsal interneurons (Lhx2/9+). (F) Control staining of E12.5 rat spinal cord double-labeled for Lhx2/9 (F″, green) and Brn3a (F‴, red) shows Lhx2/9-positive dorsal interneurons. (G-G″) dNT cells cultured for 3 days in 100 ng/ml GDF7. Most Brn3a+ cells (G) are TrkA+ (G′), indicating that they are peripheral S neurons. (Magnification: A-E′, ×10; G-G″, ×20).
Fig. 4.
Fig. 4.
GDF7 and BMPs exhibit distinct neuronal subtype-inducing profiles in dNT cultures. Dissociated E10.5 rat neural tube cultures were grown in the presence of the indicated growth factor for 3 days, then fixed and double-labeled with antibodies to Brn3a (A) or Phox2b (B). The total number of Phox2b or Brn3a expressing cells per culture is indicated. The data were pooled from six experiments with three to six cultures per experiment per concentration, although not all concentrations were tested in every experiment. (A) In BMP2, BMP6, and BMP7, the induction of Brn3a peaks at 10 ng/ml and declines with increasing concentrations, whereas that in GDF7 increases over this same concentration range (P < 0.02). (B) Phox2b expression increases with progressively higher concentrations of BMP2, BMP6, and BMP7, whereas little or no expression is seen in GDF7. P < 0.03. Error bars indicate SEM.
Fig. 5.
Fig. 5.
Transient exposure to GDF7 commits multipotent S/A progenitors to a S fate. (A-D) Cultures were preincubated with or without GDF7 (100 ng/ml) for 48 h, then washed and switched to medium with or without BMP2 (100 ng/ml) for an additional 24 h (left flow diagram) before double-labeling for Brn3a (green) and Phox2b (red). (C) BMP2 did not reduce the number of Brn3a+ neurons that develop after GDF7 washout (G→0 vs. G→ BMP). Data represent two different experiments each containing five replicates. (D) Time course of Brn3a induction by GDF7 compared with the control (“no add”). Brn3a is only weakly expressed at day 2, the time of GDF7 washout (flow diagram). (E-G) dNT cultures were preincubated with or without 100 ng/ml BMP2 for 24 h, washed, and then incubated in 100 ng/ml GDF7 for 3 additional days (right flow diagram). (E and E′) S neurons develop in cultures preincubated in control medium, and no A neurons develop. (F and F′) S neuron differentiation is strongly inhibited if the cultures are preincubated in BMP2, and primarily A neurons develop. (G) Quantification. Note the reciprocity in the number of S vs. A neurons under the two conditions.
Fig. 6.
Fig. 6.
Pleuripotent and restricted S progenitors coexist in the premigratory neural crest. Premigratory neural crest cells in the roof plate (purple) marked by expression of Gdf7 are restricted to S fates (both neuronal and glial; right). The data do not exclude the possibility that some S fate-restricted progenitors are also present outside the roof plate. GDF7 itself may play a role in S fate restriction. NT, neural tube; DA, dorsal aorta.
See this image and copyright information in PMC

References

    1. Le Douarin, N. M. & Kalcheim, C. (1999) The Neural Crest (Cambridge Univ. Press, Cambridge, U.K.).
    1. Le Douarin, N. M. (1986) Science 231, 1515-1522. - PubMed
    1. Bronner-Fraser, M. & Fraser, S. (1988) Nature 335, 161-164. - PubMed
    1. Bronner-Fraser, M. & Fraser, S. (1989) Neuron 3, 755-766. - PubMed
    1. Fraser, S. E. & Bronner-Fraser, M. E. (1991) Development (Cambridge, U.K.) 112, 913-920. - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources