Histological and Physiological Investigation of Channelrhodopsin-2 and Halorhodopsin in the Dorsal Cochlear Nucleus  

作　　者：T. Shimano B. Fyk-Kolodziej M. Asako K. Tomoda S. Bledsoe Z.H. Pan S. Molitor A.G. Holt 

机构地区：[1]Anatomy and Cell Biology Department, Wayne State University School of Medicine, Detroit, MI, 48201, USA [2]Department of Otolaryngology, Kansai Medical University, Hirakata-city, Osaka, 573-1191, Japan [3]Kresge Hearing Research Institute, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA [4]Department of Bioengineering, University of Toledo, Toledo, OH, 43606, USA

出　　处：《Journal of Otology》2011年第1期10-19,共10页中华耳科学杂志（英文版）

基　　金：supported by Ralph Wilson Foundation(to A.G.H);Capita Foundation(to A.G.H)

摘　　要：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, 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.

关 键 词：cochlear nucleus channelrhodopsin-2 HALORHODOPSIN TRACER optical control 

分 类 号：R339.1[医药卫生—人体生理学]