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Sonic hedgehog regulates the proliferation, differentiation, and migration of enteric neural crest cells in gut.

Fu M, Lui VC, Sham MH, Pachnis V, Tam PK - J. Cell Biol. (2004)

Bottom Line: The pro-neurogenic effect of glial cell line--derived neurotrophic factor (GDNF) on NCCs was abolished by Shh.In gut explants, NCCs migrated from the explants onto the adjacent substratum if GDNF was added, whereas addition of Shh abolished this migration.Our data suggest that Shh controls the proliferation and differentiation of NCCs and modulates the responsiveness of NCCs toward GDNF inductions.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, The University of Hong Kong, 21 Sassoon Rd., Pokfulam, Hong Kong, HKSAR China.

ABSTRACT
Enteric neural crest cells (NCCs) migrate and colonize the entire gut and proliferate and differentiate into neurons and glia of the enteric nervous system in vertebrate embryos. We have investigated the mitogenic and morphogenic functions of Sonic hedgehog (Shh) on enteric NCCs in cell and organ culture. Enteric NCCs expressed Shh receptor Patched and transcripts encoding the Shh signal transducer (Gli1). Shh promoted the proliferation and inhibited the differentiation of NCCs. The pro-neurogenic effect of glial cell line--derived neurotrophic factor (GDNF) on NCCs was abolished by Shh. In gut explants, NCCs migrated from the explants onto the adjacent substratum if GDNF was added, whereas addition of Shh abolished this migration. Neuronal differentiation and coalescence of neural crest--derived cells into myenteric plexuses in explants was repressed by the addition of Shh. Our data suggest that Shh controls the proliferation and differentiation of NCCs and modulates the responsiveness of NCCs toward GDNF inductions.

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Shh suppresses the differentiation of NCCs. NCC clusters were seen after 18 h of incubation, and clusters increased in size after two more days in Shh culture (A, asterisk). NCC clusters (asterisks) could also be found in GDNF culture (B) and in control culture (C; plain medium) after 40 h of incubation. Neurite projections (arrowheads) were obvious in GDNF culture (B) and in control culture (C). (D–O) NCC clusters were localized by Ret staining (red; marked with dashed lines), and proliferative cells in the clusters were detected with anti-BrdU antibody (green). BrdU+ cells were localized within the clusters (E, H, K, and N), and very few mesenchyme cells were found to be BrdU+ (E, arrowhead). NCC clusters pretreated with Shh (Q and R) or without Shh (P) were incubated with new media with (P and R) or without (Q) GDNF. Cultures were stained for TUJ1 (P–R, red). All photos were taken at the same magnification. Bars, 100 μm.
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fig4: Shh suppresses the differentiation of NCCs. NCC clusters were seen after 18 h of incubation, and clusters increased in size after two more days in Shh culture (A, asterisk). NCC clusters (asterisks) could also be found in GDNF culture (B) and in control culture (C; plain medium) after 40 h of incubation. Neurite projections (arrowheads) were obvious in GDNF culture (B) and in control culture (C). (D–O) NCC clusters were localized by Ret staining (red; marked with dashed lines), and proliferative cells in the clusters were detected with anti-BrdU antibody (green). BrdU+ cells were localized within the clusters (E, H, K, and N), and very few mesenchyme cells were found to be BrdU+ (E, arrowhead). NCC clusters pretreated with Shh (Q and R) or without Shh (P) were incubated with new media with (P and R) or without (Q) GDNF. Cultures were stained for TUJ1 (P–R, red). All photos were taken at the same magnification. Bars, 100 μm.

Mentions: NCC clusters were seen in Shh culture after 18 h of incubation, and the clusters increased in size after two more days (Fig. 4 A). NCC clusters were detected only after 3–4 d of incubation with GDNF (Fig. 4 B) and were generally smaller than those formed in the presence of Shh (compare Fig. 4 A and Fig. 4 B). Only few and relatively small clusters were observed in the culture with plain medium (Fig. 4 C, control). Cells of clusters in different treatments displayed variable morphologies. Neuronal processes (Fig. 4, arrowheads) were scarce in Shh culture, but were very prominent in the cultures with GDNF and in control (compare Fig. 4, A–C). Dissociated gut cells had been filtered through cell strainers to remove cell clumps before plating, so NCC cluster formation in culture was not the result of insufficient dissociation of gut cells. Bigger size of clusters in culture could reflect a higher proliferation of NCCs.


Sonic hedgehog regulates the proliferation, differentiation, and migration of enteric neural crest cells in gut.

Fu M, Lui VC, Sham MH, Pachnis V, Tam PK - J. Cell Biol. (2004)

Shh suppresses the differentiation of NCCs. NCC clusters were seen after 18 h of incubation, and clusters increased in size after two more days in Shh culture (A, asterisk). NCC clusters (asterisks) could also be found in GDNF culture (B) and in control culture (C; plain medium) after 40 h of incubation. Neurite projections (arrowheads) were obvious in GDNF culture (B) and in control culture (C). (D–O) NCC clusters were localized by Ret staining (red; marked with dashed lines), and proliferative cells in the clusters were detected with anti-BrdU antibody (green). BrdU+ cells were localized within the clusters (E, H, K, and N), and very few mesenchyme cells were found to be BrdU+ (E, arrowhead). NCC clusters pretreated with Shh (Q and R) or without Shh (P) were incubated with new media with (P and R) or without (Q) GDNF. Cultures were stained for TUJ1 (P–R, red). All photos were taken at the same magnification. Bars, 100 μm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2172437&req=5

fig4: Shh suppresses the differentiation of NCCs. NCC clusters were seen after 18 h of incubation, and clusters increased in size after two more days in Shh culture (A, asterisk). NCC clusters (asterisks) could also be found in GDNF culture (B) and in control culture (C; plain medium) after 40 h of incubation. Neurite projections (arrowheads) were obvious in GDNF culture (B) and in control culture (C). (D–O) NCC clusters were localized by Ret staining (red; marked with dashed lines), and proliferative cells in the clusters were detected with anti-BrdU antibody (green). BrdU+ cells were localized within the clusters (E, H, K, and N), and very few mesenchyme cells were found to be BrdU+ (E, arrowhead). NCC clusters pretreated with Shh (Q and R) or without Shh (P) were incubated with new media with (P and R) or without (Q) GDNF. Cultures were stained for TUJ1 (P–R, red). All photos were taken at the same magnification. Bars, 100 μm.
Mentions: NCC clusters were seen in Shh culture after 18 h of incubation, and the clusters increased in size after two more days (Fig. 4 A). NCC clusters were detected only after 3–4 d of incubation with GDNF (Fig. 4 B) and were generally smaller than those formed in the presence of Shh (compare Fig. 4 A and Fig. 4 B). Only few and relatively small clusters were observed in the culture with plain medium (Fig. 4 C, control). Cells of clusters in different treatments displayed variable morphologies. Neuronal processes (Fig. 4, arrowheads) were scarce in Shh culture, but were very prominent in the cultures with GDNF and in control (compare Fig. 4, A–C). Dissociated gut cells had been filtered through cell strainers to remove cell clumps before plating, so NCC cluster formation in culture was not the result of insufficient dissociation of gut cells. Bigger size of clusters in culture could reflect a higher proliferation of NCCs.

Bottom Line: The pro-neurogenic effect of glial cell line--derived neurotrophic factor (GDNF) on NCCs was abolished by Shh.In gut explants, NCCs migrated from the explants onto the adjacent substratum if GDNF was added, whereas addition of Shh abolished this migration.Our data suggest that Shh controls the proliferation and differentiation of NCCs and modulates the responsiveness of NCCs toward GDNF inductions.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, The University of Hong Kong, 21 Sassoon Rd., Pokfulam, Hong Kong, HKSAR China.

ABSTRACT
Enteric neural crest cells (NCCs) migrate and colonize the entire gut and proliferate and differentiate into neurons and glia of the enteric nervous system in vertebrate embryos. We have investigated the mitogenic and morphogenic functions of Sonic hedgehog (Shh) on enteric NCCs in cell and organ culture. Enteric NCCs expressed Shh receptor Patched and transcripts encoding the Shh signal transducer (Gli1). Shh promoted the proliferation and inhibited the differentiation of NCCs. The pro-neurogenic effect of glial cell line--derived neurotrophic factor (GDNF) on NCCs was abolished by Shh. In gut explants, NCCs migrated from the explants onto the adjacent substratum if GDNF was added, whereas addition of Shh abolished this migration. Neuronal differentiation and coalescence of neural crest--derived cells into myenteric plexuses in explants was repressed by the addition of Shh. Our data suggest that Shh controls the proliferation and differentiation of NCCs and modulates the responsiveness of NCCs toward GDNF inductions.

Show MeSH
Related in: MedlinePlus