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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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Expression of Ptc1 and Gli1 in mouse embryonic guts was analyzed by in situ hybridization. NCCs that expressed Ret were localized within the E11.5 gut mesenchyme (A and C, arrowheads). Transcripts of Ptc1 (B) and Gli1 (D) were localized within the mesenchyme. Using anti-Ret and -Ptc1 antibodies, Ptc1 protein was detected at the NCCs (asterisks) and the mesenchyme of the E11.5 gut (E–G) and E13.5 gut (H–J). Transcripts of Shh were restricted to the mucosa (K). Immunofluorescence using antibody 5E1 localized Shh protein at the mucosa and mesenchyme of the E11.5 gut (L). Staining with IgG isotype control showed a weak nonspecific signal (M). Dashed lines in L mark the gut border. m, mucosa. Photos A–G and K–M were taken at the same magnification. Photos H–J were taken at the same magnification. Bars: (A–G and K–M) 100 μm; (H–J) 50 μm.
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fig1: Expression of Ptc1 and Gli1 in mouse embryonic guts was analyzed by in situ hybridization. NCCs that expressed Ret were localized within the E11.5 gut mesenchyme (A and C, arrowheads). Transcripts of Ptc1 (B) and Gli1 (D) were localized within the mesenchyme. Using anti-Ret and -Ptc1 antibodies, Ptc1 protein was detected at the NCCs (asterisks) and the mesenchyme of the E11.5 gut (E–G) and E13.5 gut (H–J). Transcripts of Shh were restricted to the mucosa (K). Immunofluorescence using antibody 5E1 localized Shh protein at the mucosa and mesenchyme of the E11.5 gut (L). Staining with IgG isotype control showed a weak nonspecific signal (M). Dashed lines in L mark the gut border. m, mucosa. Photos A–G and K–M were taken at the same magnification. Photos H–J were taken at the same magnification. Bars: (A–G and K–M) 100 μm; (H–J) 50 μm.

Mentions: Spatio-temporal expression of Ptc1 and Gli1 in mouse embryonic guts was analyzed by in situ hybridization and immunohistochemistry. NCCs that expressed Ret were localized within the E11.5 mouse gut mesenchyme (Fig. 1, A and C). Transcripts of Ptc1 and Gli1 were localized within the mesenchyme (Fig. 1, B and D). Patched (Ptc1) immunoreactivity was localized at the E11.5 gut mesenchyme and Ret+ NCCs (Fig. 1, E–G). At E13.5, Ptc1 was detected at the mesenchyme and at the presumptive myenteric region underneath the serosa where Ret+ NCCs were found (Fig. 1, H–J). Transcripts of Shh were restricted to the mucosa of the E11.5 gut (Fig. 1 K). However, Shh protein was detected at the mucosa and mesenchyme by immunofluorescence (Fig. 1 L). The signal was strong at the mucosa, whereas the signal at the mesenchyme declined the further it was away from the region adjacent to the mucosa. These findings indicated that Shh was secreted from the mucosa into the mesenchyme forming a concentration gradient across the gut radius. Staining with mouse IgG isotype control antibody gave a weak nonspecific signal (Fig. 1 M).


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)

Expression of Ptc1 and Gli1 in mouse embryonic guts was analyzed by in situ hybridization. NCCs that expressed Ret were localized within the E11.5 gut mesenchyme (A and C, arrowheads). Transcripts of Ptc1 (B) and Gli1 (D) were localized within the mesenchyme. Using anti-Ret and -Ptc1 antibodies, Ptc1 protein was detected at the NCCs (asterisks) and the mesenchyme of the E11.5 gut (E–G) and E13.5 gut (H–J). Transcripts of Shh were restricted to the mucosa (K). Immunofluorescence using antibody 5E1 localized Shh protein at the mucosa and mesenchyme of the E11.5 gut (L). Staining with IgG isotype control showed a weak nonspecific signal (M). Dashed lines in L mark the gut border. m, mucosa. Photos A–G and K–M were taken at the same magnification. Photos H–J were taken at the same magnification. Bars: (A–G and K–M) 100 μm; (H–J) 50 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172437&req=5

fig1: Expression of Ptc1 and Gli1 in mouse embryonic guts was analyzed by in situ hybridization. NCCs that expressed Ret were localized within the E11.5 gut mesenchyme (A and C, arrowheads). Transcripts of Ptc1 (B) and Gli1 (D) were localized within the mesenchyme. Using anti-Ret and -Ptc1 antibodies, Ptc1 protein was detected at the NCCs (asterisks) and the mesenchyme of the E11.5 gut (E–G) and E13.5 gut (H–J). Transcripts of Shh were restricted to the mucosa (K). Immunofluorescence using antibody 5E1 localized Shh protein at the mucosa and mesenchyme of the E11.5 gut (L). Staining with IgG isotype control showed a weak nonspecific signal (M). Dashed lines in L mark the gut border. m, mucosa. Photos A–G and K–M were taken at the same magnification. Photos H–J were taken at the same magnification. Bars: (A–G and K–M) 100 μm; (H–J) 50 μm.
Mentions: Spatio-temporal expression of Ptc1 and Gli1 in mouse embryonic guts was analyzed by in situ hybridization and immunohistochemistry. NCCs that expressed Ret were localized within the E11.5 mouse gut mesenchyme (Fig. 1, A and C). Transcripts of Ptc1 and Gli1 were localized within the mesenchyme (Fig. 1, B and D). Patched (Ptc1) immunoreactivity was localized at the E11.5 gut mesenchyme and Ret+ NCCs (Fig. 1, E–G). At E13.5, Ptc1 was detected at the mesenchyme and at the presumptive myenteric region underneath the serosa where Ret+ NCCs were found (Fig. 1, H–J). Transcripts of Shh were restricted to the mucosa of the E11.5 gut (Fig. 1 K). However, Shh protein was detected at the mucosa and mesenchyme by immunofluorescence (Fig. 1 L). The signal was strong at the mucosa, whereas the signal at the mesenchyme declined the further it was away from the region adjacent to the mucosa. These findings indicated that Shh was secreted from the mucosa into the mesenchyme forming a concentration gradient across the gut radius. Staining with mouse IgG isotype control antibody gave a weak nonspecific signal (Fig. 1 M).

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