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We have delivered viral vectors containing either Chop2 fused with GFP, Channelrhodopsin-2 (ChR2), or Halorhodopsin (HaloR) fused with mCherry (to form light gated cation channels or chloride pumps, respectively), into the dorsal cochlear nucleus (DCN). One to eighteen months later we examined the CN and inferior colliculus (IC) for evidence of virally transfected cells and processes. Production of ChR2 and HaloR was observed throughout the DCN. Rhodopsin localization within neurons was determined, with elongate, fusiform and giant cells identified based on morphology and location within the DCN. Production of ChR2 and HaloR was found at both the injection site as well as in regions projecting to and from the DCN. Light driven neuronal activity in the DCN was dependent upon the wavelength and intensity of the light, with only the appropriate wavelength resulting in activation and higher intensity light resulting in more neuronal activity. Transfecting cells via viral delivery of rhodopsins can be useful as a tract tracer and as a neuronal marker to delineate pathways. In the future rhodopsin delivery and activation may be developed as an alternative to electrical stimulation of neurons.
We have delivered viral vectors containing either Chop2 fused with GFP, Channel rhodopsin-2 (ChR2), or Halorhodopsin (HaloR) fused with mCherry (to form light gated cation channels or chloride pumps, respectively) into the dorsal cochlear nucleus (DCN). One to eighteen months later we observed the CN and inferior colliculus (IC) for evidence of virally transfected cells and processes. Production of ChR2 and HaloR was observed throughout the DCN. Rhodopsin localization within neurons was determined, with elongate, fusiform and giant cells identified based on morphology and location within the DCN. Production of ChR2 and HaloR was found at both both injection site as well as in regions projecting to and from the DCN. Light driven neuronal activity in the DCN was dependent upon the wavelength and intensity of the light , with only the Appropriate wavelength resulting in activation and higher intensity light resulting in more neuronal activity. Transfecting cells via viral delivery of rhodop sins can be useful as a tracer and as a neuronal marker to delineate pathways. In the future rhodopsin delivery and activation may be developed as an alternative to electrical stimulation of neurons.