Abstract
The Cre/loxP system has been widely used to generate tissue-specific gene knockout mice. Inducible (Tet-off) Osx-GFP::Cre (Osx-Cre) mouse line that targets osteoblasts is widely used in the bone research field. In this study, we investigated the effect of Osx-Cre on craniofacial bone development. We found that newborn Osx-Cre mice showed severe hypomineralization in parietal, frontal, and nasal bones as well as the coronal sutural area when compared to control mice. As the mice matured, the intramembranous bone hypomineralization phenotype became less severe. The major hypomineralization defect in parietal, frontal, and nasal bones had mostly disappeared by postnatal day 21, but the defect in sutural areas persisted. Importantly, Doxycycline treatment eliminated cranial bone defects at birth which indicates that Cre expression may be responsible for the phenotype. In addition, we showed that the primary calvarial osteoblasts isolated from neonatal Osx-Cre mice had comparable differentiation ability compared to their littermate controls. This study reinforces the idea that Cre-positive litter mates are indispensable controls in studies using conditional gene deletion.
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Acknowledgments
We thank Dr. Andrew McMahon for providing the Osx-Cre mice; Dr. Taylor Snider for critically reading and editing the manuscript. This study was funded by the National Institute Of Arthritis And Musculoskeletal And Skin Diseases of the National Institutes of Health under Award Number R01AR062030 (to F.L.) and R01DE020843 (to Y.M.).
Conflict of Interest
Dr. Fei Liu reports Grant from NIH during the conduct of the study. Dr. Yuji Mishina reports grants from NIH/NIDCR, personal fees from NIH/NICHD, others from NIH/NIEHS, personal fees from Shriners Hospital, grants from Department of Defense during the conduct of the study. Dr. Li Wang has nothing to disclose.
Human and Animal Rights and Informed Consent
This article does not contain any studies with human subjects performed by any of the authors. All mice procedures used in this study were approved by the Institutional Animal Care and Use Committee at the University of Michigan.
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Wang, L., Mishina, Y. & Liu, F. Osterix-Cre Transgene Causes Craniofacial Bone Development Defect. Calcif Tissue Int 96, 129–137 (2015). https://doi.org/10.1007/s00223-014-9945-5
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DOI: https://doi.org/10.1007/s00223-014-9945-5