Mechanosensation, the transduction of mechanical force into electrochemical signals, allows organisms to detect touch and sound, to register movement and gravity, and to sense changes in cell volume and shape. The hair cells of the mammalian inner ear are the mechanosensors for the detection of sound and head movement. The analysis of gene function in hair cells has been hampered by the lack of an efficient gene transfer method. Here we describe a method termed injectoporation that combines tissue microinjection with electroporation to express cDNAs and shRNAs in mouse cochlear hair cells. Injectoporation allows for gene transfer into dozens of hair cells, and it is compatible with the analysis of hair cell function using imaging approaches and electrophysiology. Tissue dissection and injectoporation can be carried out within a few hours, and the tissue can be cultured for days for subsequent functional analyses.
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Improved calcium sensor GCaMP-X overcomes the calcium channel perturbations induced by the calmodulin in GCaMP
Nature Communications Open Access 17 April 2018
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We thank J. Bartles (Northwestern University) for espin-EGFP. This research was supported by funding from the US National Institutes of Health (NIH) (to U.M., DC005965 and DC007704), the Dorris Neuroscience Center (to U.M.), the Deutsche Forschungsgemeinschaft (to T.W.) and the Bundy foundation (to W.X.).
The authors declare no competing financial interests.
Integrated supplementary information
Supplementary Figure 1 Gene transfer into supporting cells by injectoporation.
A diagram of a cross section through the organ of Corti is shown as well as the EGFP fluorescence signal in the indicated cell types. Scale bars: 10 μm.
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Xiong, W., Wagner, T., Yan, L. et al. Using injectoporation to deliver genes to mechanosensory hair cells. Nat Protoc 9, 2438–2449 (2014). https://doi.org/10.1038/nprot.2014.168
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