Generation of a MOR‐CreER knock‐in mouse line to study cells and neural circuits involved in mu opioid receptor signaling
Generation of a MOR‐CreER knock‐in mouse line to study cells and neural circuits involved in mu opioid receptor signaling
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DOI：
10.1002/dvg.23341
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发表时间：
2020-01
期刊：
影响因子：
1.5
通讯作者：
T. Okunomiya;H. Hioki;Chika Nishimura;Satoshi Yawata;I. Imayoshi;R. Kageyama;R. Takahashi;Dai Watanabe
中科院分区：
文献类型：
--
作者：
T. Okunomiya;H. Hioki;Chika Nishimura;Satoshi Yawata;I. Imayoshi;R. Kageyama;R. Takahashi;Dai Watanabe
Mu阿片受体（Mu opioid receptor， MOR）参与多种脑功能，如疼痛调节、奖励加工和成瘾行为，并介导吗啡和其他阿片化合物的主要药理作用。为了获得MOR‐表达细胞的遗传途径，并研究MOR‐信号传导的生理和病理作用，我们构建了MOR‐CreER敲入小鼠系，其中Oprm1基因的停止密码子被编码T2A肽和他莫昔芬（Tm）诱导的Cre重组酶的DNA片段所取代。我们发现，MOR - CreER等位基因在成人神经系统中表达MOR的神经元的一个离散亚型中经历了Tm依赖性重组，这些神经元包括嗅球、大脑皮层、纹状体、海马、杏仁核、丘脑、下丘脑、脚间核、上丘和下丘、导水管周围灰质、臂旁核、耳蜗核、中缝核、脑桥和延髓网状结构、歧见核、孤立核、脊髓和背根神经节。MOR‐CreER小鼠系与Cre依赖性腺相关病毒载体结合，可以在MOR‐富集的纹状体中进行稳健的基因操作。此外，在产前发育期间，Tm治疗可有效诱导Cre介导的重组。因此，MOR‐CreER小鼠是研究发育和成熟神经组织中MOR‐表达细胞条件基因操作的有力工具。
Mu opioid receptor (MOR) is involved in various brain functions, such as pain modulation, reward processing, and addictive behaviors, and mediates the main pharmacologic effects of morphine and other opioid compounds. To gain genetic access to MOR‐expressing cells, and to study physiological and pathological roles of MOR signaling, we generated a MOR‐CreER knock‐in mouse line, in which the stop codon of the Oprm1 gene was replaced by a DNA fragment encoding a T2A peptide and tamoxifen (Tm)‐inducible Cre recombinase. We show that the MOR‐CreER allele undergoes Tm‐dependent recombination in a discrete subtype of neurons that express MOR in the adult nervous system, including the olfactory bulb, cerebral cortex, striosome compartments in the striatum, hippocampus, amygdala, thalamus, hypothalamus, interpeduncular nucleus, superior and inferior colliculi, periaqueductal gray, parabrachial nuclei, cochlear nucleus, raphe nuclei, pontine and medullary reticular formation, ambiguus nucleus, solitary nucleus, spinal cord, and dorsal root ganglia. The MOR‐CreER mouse line combined with a Cre‐dependent adeno‐associated virus vector enables robust gene manipulation in the MOR‐enriched striosomes. Furthermore, Tm treatment during prenatal development effectively induces Cre‐mediated recombination. Thus, the MOR‐CreER mouse is a powerful tool to study MOR‐expressing cells with conditional gene manipulation in developing and mature neural tissues.