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A Sox10rtTA/+ Mouse Line Allows for Inducible Gene Expression in the Auditory and Balance Organs of the Inner Ear

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Abstract

Genetic mouse models provide invaluable tools for discerning gene function in vivo. Tetracycline-inducible systems (Tet-On/Off) provide temporal and cell-type specific control of gene expression, offering an alternative or even complementary approach to existing Cre/LoxP systems. Here we characterized a Sox10rtTA/+ knock-in mouse line which demonstrates inducible reverse tetracycline trans-activator (rtTA) activity and Tet-On transgene expression in the inner ear following induction with the tetracycline derivative doxycycline (Dox). These Sox10rtTA/+ mice do not exhibit any readily observable developmental or hearing phenotypes, and actively drive Tet-On transgene expression in Sox10 expressing cells in the inner ear. Sox10rtTA/+ activity was revealed by multiple Tet-On reporters to be nearly ubiquitous throughout the membranous labyrinth of the developing inner ear, and notably absent from hair cells, tympanic border cells, and ganglion neurons following postnatal Dox inductions. Interestingly, Dox-induced Sox10rtTA/+ activity declined with induction age, where Tet-On reporters became uninducible in adult cochlear epithelium. Co-administration of the loop diuretic furosemide was able to rescue Dox-induced reporter expression, though this method also caused significant cochlear hair cell loss. Surprisingly, Sox10rtTA/+ driven reporter expression in the cochlea persists for at least 54 days after cessation of neonatal induction, presumably due to the persistence of Dox within inner ear tissues. These findings highlight the utility of the Sox10rtTA/+ mouse line as a powerful tool for functional genetic studies of the auditory and balance organs in vivo, but also reveal some important considerations that must be adequately controlled for in future studies that rely upon Tet-On/Off systems.

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Acknowledgments

The authors would like to thank Lingli Zhang and Mario Sauceda for assistance with genotyping and Dr. Brandon Cox for helpful comments in the preparation of this manuscript. This research was supported by funding from the National Institutes of Health (grants DC006471 (J.Z.), P30CA21765 (St. Jude)), American Lebanese Syrian Associated Charities (ALSAC) of St. Jude Children’s Research Hospital, the Office of Naval Research (grants N000140911014, N000141210191, N000141210775 (J.Z.)), the National Organization for Hearing Research (NOHR) Foundation (B.W.), the Hearing Health Foundation (Emerging Research Grant, B.W.), and The Hartwell Foundation (Individual Biomedical Research Award, J.Z.).

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  1. Department of Developmental Neurobiology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN, 38105, USA

    Bradley J. Walters & Jian Zuo

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Correspondence to Jian Zuo.

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Walters, B.J., Zuo, J. A Sox10rtTA/+ Mouse Line Allows for Inducible Gene Expression in the Auditory and Balance Organs of the Inner Ear. JARO 16, 331–345 (2015). https://doi.org/10.1007/s10162-015-0517-9

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