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Targeted expression of step-function opsins in transgenic rats for optogenetic studies

Rats are excellent animal models for experimental neuroscience. However, the application of optogenetics in rats has been hindered because of the limited number of established transgenic rat strains. To accomplish cell-type specific targeting of an optimized optogenetic molecular tool, we generated...

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Detalles Bibliográficos
Autores principales: Igarashi, Hiroyuki, Ikeda, Keiko, Onimaru, Hiroshi, Kaneko, Ryosuke, Koizumi, Kyo, Beppu, Kaoru, Nishizawa, Kayo, Takahashi, Yukari, Kato, Fusao, Matsui, Ko, Kobayashi, Kazuto, Yanagawa, Yuchio, Muramatsu, Shin-Ichi, Ishizuka, Toru, Yawo, Hiromu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5882906/
https://www.ncbi.nlm.nih.gov/pubmed/29615713
http://dx.doi.org/10.1038/s41598-018-23810-8
Descripción
Sumario:Rats are excellent animal models for experimental neuroscience. However, the application of optogenetics in rats has been hindered because of the limited number of established transgenic rat strains. To accomplish cell-type specific targeting of an optimized optogenetic molecular tool, we generated ROSA26/CAG-floxed STOP-ChRFR(C167A)-Venus BAC rats that conditionally express the step-function mutant channelrhodopsin ChRFR(C167A) under the control of extrinsic Cre recombinase. In primary cultured cortical neurons derived from this reporter rat, only Cre-positive cells expressing ChRFR(C167A) became bi-stable, that is, their excitability was enhanced by blue light and returned to the baseline by yellow~red light. In bigenic pups carrying the Phox2B-Cre driver, ChRFR(C167A) was specifically expressed in the rostral parafacial respiratory group (pFRG) in the medulla, where endogenous Phox2b immunoreactivity was detected. These neurons were sensitive to blue light with an increase in the firing frequency. Thus, this transgenic rat actuator/reporter system should facilitate optogenetic studies involving the effective in vivo manipulation of the activities of specific cell fractions using light of minimal intensity.