Abstract
The Smad2/3 pathway plays a key role in mediating TGF-β1 inhibition of branching morphogenesis and induction of connective tissue growth factor (CTGF) expression in embryonic lungs. Because a number of cell-specific interactions have been described between TGF-β1-driven Smad signaling and the c-Jun N-terminal kinase (JNK) pathway, we have investigated the effects of JNK inhibition on TGF-β1 activation of Smad2, inhibition of branching, induction of CTGF expression, and apoptosis in mouse embryonic lung explants. Mouse embryonic day 12.5 (E12.5) lung explants were treated with TGF-β1 in the presence or absence of a specific pharmacologic JNK inhibitor (SP600125) and a specific JNK peptide inhibitor (JNKI). We found that TGF-β1 activated the JNK pathway by stimulating c-Jun phosphorylation, which was blocked by JNK inhibitors. Treatment with SP600125 stimulated Smad2 phosphorylation and enhanced TGF-β1-induced Smad2 phosphorylation. Treatment with JNK inhibitors also decreased normal branching morphogenesis and induced CTGF expression as well as augmented TGF-β1 inhibition of branching and induction of CTGF expression. Furthermore, JNK inhibition-induced apoptosis. Our results demonstrate that inhibition of the JNK pathway promotes TGF-β1-driven Smad2 responses in lung branching morphogenesis. These data suggest that the JNK pathway may antagonize TGF-β1 dependent Smad2 signaling during mouse embryonic lung development.
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Abbreviations
- AP-1:
-
activating-protein-1
- CTGF:
-
connective tissue growth factor
- JNK:
-
c-Jun NH2-terminal kinase
- JNKI:
-
JNK peptide inhibitor
- JNKC:
-
JNK peptide control
- MAPK:
-
mitogen-activated protein kinase
- MKK:
-
MAPK kinase
- p-c-Jun:
-
phosphorylated c-Jun
- p-Smad2:
-
phosphorylated smad2
- TβRI:
-
TGF-β receptor I
- TβRII:
-
TGF-β receptor II
- t-c-Jun:
-
total c-Jun
- t-Smad2:
-
total Smad2
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Supported, in part, by grant K08 HD046582 from NIH, by Project NewBorn University of Miami, and by a grant from the Bank of American Charitable Foundation, Inc.
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Wu, S., Kasisomayajula, K., Peng, J. et al. Inhibition of JNK Enhances TGF-β1-Activated Smad2 Signaling in Mouse Embryonic Lung. Pediatr Res 65, 381–386 (2009). https://doi.org/10.1203/PDR.0b013e3181991c67
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DOI: https://doi.org/10.1203/PDR.0b013e3181991c67
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