Bone morphogenetic protein type I receptor inhibition induces cleft palate associated with micrognathia and cleft lower lip in mice

Yongzhen Lai; Changfu Xie; Shixian Zhang; Guowu Gan; Di Wu; Weihui Chen

doi:10.1002/bdra.23504

Bone morphogenetic protein type I receptor inhibition induces cleft palate associated with micrognathia and cleft lower lip in mice

Birth Defects Res A Clin Mol Teratol. 2016 Jul;106(7):612-23. doi: 10.1002/bdra.23504. Epub 2016 May 6.

Authors

Yongzhen Lai¹, Changfu Xie¹, Shixian Zhang¹, Guowu Gan¹, Di Wu¹, Weihui Chen^{1

2}

Affiliations

¹ Department of Oral and Maxillofacial Surgery, Union Hospital, Fujian Medical University, Fuzhou, P. R. China.
² Stomatological Research Institute, Fujian Medical University, Fuzhou, Fujian Province, P. R. China.

PMID: 27150428
DOI: 10.1002/bdra.23504

Abstract

Background: Gain-of- and loss-of-function studies have demonstrated that changes in bone morphogenetic protein (BMP) signaling during embryo development cause craniofacial malformations, including cleft palate. It remains uncertain whether BMP signaling could be targeted pharmacologically to affect craniofacial morphogenesis.

Methods: Pregnant C57Bl/6J mice were treated with the BMP type I receptor inhibitor LDN-193189 at the dose of 3, 6, or 9 mg/kg twice a day by intraperitoneal injection from embryonic day 10.5 (E10.5) to E15.5. At E16.5, embryos were investigated by facial measurement analysis and histology to determine the optimal concentration for malformation. Subsequent embryonic phenotypes were analyzed in detail by histology, whole-mount skeletal staining, micro-computed tomography, and palatal organic culture. We further used immunohistochemistry to analyze protein expression of the BMP-mediated canonical and noncanonical signaling components.

Results: The optimal concentration of LDN-193189 was determined to be 6 mg/kg. In utero, LDN-193189 exposures induced partial clefting of the anterior palate or complete cleft palate, which was attributed to a reduced cell proliferation rate in the secondary palate, and delayed palatal elevation caused by micrognathia. Analysis of signal transduction in palatal shelves at E12.5 and E13.5 identified a significant reduction of BMP/Smad signaling (p-Smad1/5/8) and unchanged BMP noncanonical signaling (p-p38, p-Erk1/2) after treatment with LDN-193189.

Conclusion: The results of this study indicate that LDN-193189 can be used to manipulate BMP signaling by selectively targeting the BMP/Smad signaling pathway to affect palatal morphogenesis and produce phenotypes mimicking those caused by genetic mutations. This work established a novel mouse model for teratogen-induced cleft palate. Birth Defects Research (Part A) 106:612-623, 2016. © 2016 Wiley Periodicals, Inc.

Keywords: BMP type I receptor; LDN-193189; cleft lower lip; cleft palate; micrognathia.

MeSH terms

Animals
Bone Morphogenetic Protein Receptors, Type I / antagonists & inhibitors*
Bone Morphogenetic Protein Receptors, Type I / metabolism*
Cleft Palate* / chemically induced
Cleft Palate* / metabolism
Male
Mice
Pyrazoles / adverse effects*
Pyrazoles / pharmacology
Pyrimidines / adverse effects*
Pyrimidines / pharmacology
Signal Transduction / drug effects*
X-Ray Microtomography

Substances

LDN 193189
Pyrazoles
Pyrimidines
Bone Morphogenetic Protein Receptors, Type I