Decreased expression of Wnt9b and Ube2ql1 in the face of CL/Fr mouse embryos at E11.5 based on microarray analysis

Abstract

The CL/Fr mouse demonstrates heritable bilateral and unilateral cleft lip and palate (CLP) at a rate of approximately 35%, generally above the background “A” strain mouse. Using classical mouse breeding strategies, it has been suggested that at least two disease loci, clf1 and clf2, are involved in the defect and candidate genes have been identified. Additionally, gene‐targeting analyses strongly suggest that Wnt9b contributes to CLP in the “A” strain mice. The aim of this study was to test the expression of clf1 and clf2 candidate genes in the facial prominences of CL/Fr embryos, utilizing microarray analysis. Medial nasal, lateral nasal, and maxillary prominences of phenotypically normal as well as cleft E11.5 CL/Fr mice were dissected and RNA was extracted using standard techniques for Agilent‐microarray protocol. Results indicate that expression of the clf1 candidate gene, Wnt9b, and the clf2 candidate gene, Ube2ql1, are significantly reduced (−3.11 and −1.83 fold, respectively) in the CL/Fr cleft tissues, suggesting these genes may be involved in the CLP mutation in CL/Fr mice. Future gene expression studies through quantitative RT‐PCR and regional expression analyses through immunohistochemistry and whole mount in situ hybridization will be performed to further test the expression of these clf1 and clf2 candidate genes.Grant Funding Source: NIH/NCRR 5P20RR024206 (S.J.S) and R01‐DK‐064752 (SL)