Transfection of mouse cochlear explants by electroporation

National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland, USA.
Current protocols in neuroscience / editorial board, Jacqueline N. Crawley ... [et al.] 04/2010; Chapter 4:Unit 4.34.1-10. DOI: 10.1002/0471142301.ns0434s51
Source: PubMed


The sensory epithelium of the mammalian inner ear, also referred to as the organ of Corti, is a remarkable structure comprised of highly ordered rows of mechanosensory hair cells and non-sensory supporting cells located within the coiled cochlea. This unit describes an in vitro explant culture technique that can be coupled with gene transfer via electroporation to study the effects of altering gene expression during development of the organ of Corti. While the protocol is largely focused on embryonic cochlea, the same basic protocol can be used on cochleae from mice as old as P5.

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    • "epithelial explants with plasmid encoding dsRed driven by a constitutive (CMV) promoter. With the electroporation method used, the majority of transfected cells observed in P1.5 cochlear epithelial samples were located in the GER, medial to the sensory cell region, as has been reported previously (Zheng and Gao, 2000; Jones et al., 2006; Driver and Kelley, 2010; Zhao et al., 2011). Cells lateral to the HCs or HCs themselves were occasionally transfected, but this was rare. "
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