The mammalian inner ear forms from a thickened patch of head ectoderm called the otic placode. The placodal ectoderm invaginates to form a cup whose edges cinch together to establish a fluid-filled sac called the otic vesicle or otocyst. The progenitor cells lining the otocyst lumen will give rise to sensory and non-sensory cells of the inner ear. These formative stages of inner ear development are initiated during the first week of postimplantation embryonic development in the mouse. The inaccessibility of the inner ear in utero has hampered efforts to gain insight into the molecular mechanisms regulating essential developmental processes. An experimental embryological method to misexpress genes in the developing mammalian inner ear is presented. Expression plasmid encoding a gene of interest is microinjected through the uterine wall into the lumen of the otocyst and electroporated into otic epithelial progenitor cells. Downstream analysis of the transfected embryonic or postnatal inner ear is then conducted to gain insight into gene function.
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Acknowledgment
We thank Tomomi Fukuchi-Shimogori and Elizabeth Grove for demonstrating transuterine microinjection techniques and Donna M. Fekete for inspired postdoctoral mentorship (to JB). Funding from the National Institute on Deafness and Other Communication Disorders and the McKnight Fund for Neuroscience is gratefully appreciated.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Brigande, J.V., Gubbels, S.P., Woessner, D.W., Jungwirth, J.J., Bresee, C.S. (2009). Electroporation-Mediated Gene Transfer to the Developing Mouse Inner Ear. In: Sokolowski, B. (eds) Auditory and Vestibular Research. Methods in Molecular Biology™, vol 493. Humana Press. https://doi.org/10.1007/978-1-59745-523-7_8
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DOI: https://doi.org/10.1007/978-1-59745-523-7_8
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