Progenitor Cells

Volume 916 of the series Methods in Molecular Biology pp 127-139


Tol2-Mediated Gene Transfer and In Ovo Electroporation of the Otic Placode: A Powerful and Versatile Approach for Investigating Embryonic Development and Regeneration of the Chicken Inner Ear

  • Stephen FreemanAffiliated withUCL Ear Institute, University College London
  • , Elena ChrysostomouAffiliated withUCL Ear Institute, University College London
  • , Koichi KawakamiAffiliated withDivision of Molecular and Developmental Biology, National Institute of Genetics
  • , Yoshiko TakahashiAffiliated withGraduate School of Biological Sciences, Nara Institute of Science and Technology (NAIST)
  • , Nicolas DaudetAffiliated withUCL Ear Institute, University College London Email author 

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The vertebrate inner ear is composed of several specialized epithelia containing mechanosensory “hair” cells, sensitive to sound and head movements. In mammals, the loss of hair cells for example during aging or after noise trauma is irreversible and results in permanent sensory deficits. By contrast, avian, fish, and amphibians can efficiently regenerate lost hair cells following trauma. The chicken inner ear is a classic model system to investigate the cellular and molecular mechanisms of inner ear development and regeneration, yet it suffered until recently from a relative lack of flexible tools for genetic studies. With the introduction of in ovo electroporation and of Tol2 transposon vectors for gene transfer in avian cells, the field of experimental possibilities has now expanded significantly in this model. Here we provide a general protocol for in ovo electroporation of the chicken otic placode and illustrate how this approach, combined with Tol2 vectors, can be used to drive long-term and inducible gene expression in the embryonic chicken inner ear. This method will be particularly useful to investigate the function of candidate genes regulating progenitor cell behavior and sensory cell differentiation in the inner ear.

Key words

Inner ear Otic placode Hair cell Chicken embryo Electroporation Transfection Tol2 transposon Development Regeneration