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

被引：14

作者：

Igarashi, Hiroyuki ^{[1
,12
]}

Ikeda, Keiko ^{[2
,3
]}

Onimaru, Hiroshi ^{[4
]}

Kaneko, Ryosuke ^{[5
]}

Koizumi, Kyo ^{[6
]}

Beppu, Kaoru ^{[7
]}

Nishizawa, Kayo ^{[8
]}

Takahashi, Yukari ^{[9
]}

Kato, Fusao ^{[9
]}

Matsui, Ko ^{[7
]}

Kobayashi, Kazuto ^{[8
]}

Yanagawa, Yuchio ^{[5
]}

Muramatsu, Shin-Ichi ^{[10
,11
]}

Ishizuka, Toru ^{[6
]}

Yawo, Hiromu ^{[1
,6
]}

机构：

[1] Tohoku Univ, Grad Sch Med, Dept Physiol & Pharmacol, Sendai, Miyagi, Japan

[2] Int Univ Hlth & Welf, Dept Physiol, Narita, Chiba, Japan

[3] Jichi Med Univ, Ctr Mol Med, Div Biol, Shimotsuke, Tochigi, Japan

[4] Showa Univ, Sch Med, Dept Physiol, Shinagawa Ku, Tokyo, Japan

[5] Gunma Univ, Grad Sch Med, Dept Genet & Behav Neurosci, Maebashi, Gunma, Japan

[6] Tohoku Univ, Grad Sch Life Sci, Dept Dev Biol & Neurosci, Sendai, Miyagi, Japan

[7] Tohoku Univ, Grad Sch Life Sci, Dept Super Network Brain Physiol, Sendai, Miyagi, Japan

[8] Fukushima Med Univ, Sch Med, Inst Biomed Sci, Dept Mol Genet, Fukushima, Japan

[9] Jikei Univ, Sch Med, Dept Neurosci, Tokyo, Japan

[10] Jichi Med Sch, Div Neurol, Shimotsuke, Tochigi, Japan

[11] Univ Tokyo, Inst Med Sci, Ctr Gene & Cell Therapy, Tokyo, Japan

[12] Japan Soc Promot Sci, Tokyo, Japan

来源：

SCIENTIFIC REPORTS | 2018年 / 8卷

关键词：

RESPIRATORY RHYTHM GENERATION; FLUORESCENT PROTEIN; NEURONS; SYSTEM; PHOX2B; MICE; CHANNELRHODOPSIN-1; MANIPULATION; ACTIVATION; MEDULLA;

D O I：

10.1038/s41598-018-23810-8

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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 similar to 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.

引用

页数：12

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