Abstract
Bronchopulmonary dysplasia (BPD) is the most common complication of preterm birth characterized by blunted post-natal lung development. BPD can be modelled in mice by exposure of newborn mouse pups to elevated oxygen levels. Little is known about the mechanisms of perturbed lung development associated with BPD. The advent of transgenic mice, where genetic rearrangements can be induced in particular cell-types at particular time–points during organogenesis, have great potential to explore the pathogenic mechanisms at play during arrested lung development. Many inducible, conditional transgenic technologies available rely on the application of the estrogen-receptor modulator, tamoxifen. While tamoxifen is well-tolerated and has been widely employed in adult mice, or in healthy developing mice; tamoxifen is not well-tolerated in combination with hyperoxia, in the most widely-used mouse model of BPD. To address this, we set out to establish a safe and effective tamoxifen dosing regimen that can be used in newborn mouse pups subjected to injurious stimuli, such as exposure to elevated levels of environmental oxygen. Our data reveal that a single intraperitoneal dose of tamoxifen of 0.2 mg applied to newborn mouse pups in 10 μl Miglyol vehicle was adequate to successfully drive Cre recombinase-mediated genome rearrangements by the fifth day of life, in a murine model of BPD. The number of recombined cells was comparable to that observed in regular tamoxifen administration protocols. These findings will be useful to investigators where tamoxifen dosing is problematic in the background of injurious stimuli and mouse models of human and veterinary disease.
References
Acharya A, Baek ST, Banfi S, Eskiocak B, Tallquist MD (2011) Efficient inducible Cre-mediated recombination in Tcf21 cell lineages in the heart and kidney. Genesis 49:870–877. doi:10.1002/dvg.20750
Burri PH (2006) Structural aspects of postnatal lung development–alveolar formation and growth. Biol Neonate 89:313–322. doi:10.1159/000092868
Casper KB, Jones K, McCarthy KD (2007) Characterization of astrocyte-specific conditional knockouts. Genesis 45:292–299. doi:10.1002/dvg.20287
Imayoshi I, Sakamoto M, Yamaguchi M, Mori K, Kageyama R (2010) Essential roles of Notch signaling in maintenance of neural stem cells in developing and adult brains. J Neurosci 30:3489–3498. doi:10.1523/JNEUROSCI.4987-09.2010
Jensen EA, Schmidt B (2014) Epidemiology of bronchopulmonary dysplasia. Birth Defects Res A Clin Mol Teratol 100:145–157. doi:10.1002/bdra.23235
Madisen L et al (2010) A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci 13:133–140. doi:10.1038/nn.2467
Mižíková I et al (2015) Collagen and elastin cross-linking is altered during aberrant late lung development associated with hyperoxia. Am J Physiol Lung Cell Mol Physiol 308:L1145–L1158. doi:10.1152/ajplung.00039.2015
Monvoisin A, Alva JA, Hofmann JJ, Zovein AC, Lane TF, Iruela-Arispe ML (2006) VE-cadherin-CreERT2 transgenic mouse: a model for inducible recombination in the endothelium. Dev Dyn 235:3413–3422. doi:10.1002/dvdy.20982
Morrisey EE, Hogan BL (2010) Preparing for the first breath: genetic and cellular mechanisms in lung development. Dev Cell 18:8–23. doi:10.1016/j.devcel.2009.12.010
Morrisey EE et al (2013) Molecular determinants of lung development. Ann Am Thorac Soc 10:S12–S16. doi:10.1513/AnnalsATS.201207-036OT
Northway WH Jr, Rosan RC, Porter DY (1967) Pulmonary disease following respirator therapy of hyaline-membrane disease. Bronchopulmonary dysplasia. N Engl J Med 276:357–368. doi:10.1056/NEJM196702162760701
Ntokou A et al (2015) Characterization of the platelet-derived growth factor receptor-alpha-positive cell lineage during murine late lung development. Am J Physiol Lung Cell Mol Physiol 309:L942–L958. doi:10.1152/ajplung.00272.2014
Rawlins EL, Perl AK (2012) The a”MAZE”ing world of lung-specific transgenic mice. Am J Respir Cell Mol Biol 46:269–282. doi:10.1165/rcmb.2011-0372PS
Silva DM, Nardiello C, Pozarska A, Morty RE (2015) Recent advances in the mechanisms of lung alveolarization and the pathogenesis of bronchopulmonary dysplasia. Am J Physiol Lung Cell Mol Physiol 309:L1239–L1272. doi:10.1152/ajplung.00268.2015
Acknowledgments
This study was supported by the Max Planck Society (J. R.-C., J.A.R.C., W.S., K.A. and R.E.M.); Rhön Klinikum AG grant Fl_66 (R.M.) and Fl_71 (K.A.); the Federal Ministry of Higher Education, Research and the Arts of the State of Hessen LOEWE Programme (all except M.D.T.); the German Center for Lung Research (Deutsches Zentrum für Lungenforschung, DZL; all except M.D.T.); and the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG) through Excellence Cluster 147 (all except M.D.T.) and individual grant Mo 1789/1 (R.M.); and National Heart, Lung, and Blood Institute grants HL074257 and HL100401 (M.D.T.).
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Ruiz-Camp, J., Rodríguez-Castillo, J.A., Herold, S. et al. Tamoxifen dosing for Cre-mediated recombination in experimental bronchopulmonary dysplasia. Transgenic Res 26, 165–170 (2017). https://doi.org/10.1007/s11248-016-9987-8
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DOI: https://doi.org/10.1007/s11248-016-9987-8