首页> 美国卫生研究院文献>Journal of Histochemistry and Cytochemistry >Distribution of MT1 Melatonin Receptor Promoter-Driven RFP Expression in the Brains of BAC C3H/HeN Transgenic Mice
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Distribution of MT1 Melatonin Receptor Promoter-Driven RFP Expression in the Brains of BAC C3H/HeN Transgenic Mice

机译:BAC C3H / HeN转基因小鼠脑中MT1褪黑素受体启动子驱动的RFP表达的分布

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摘要

The pineal hormone melatonin activates two G-protein coupled receptors (MT1 and MT2) to regulate in part biological functions. The MT1 and MT2 melatonin receptors are heterogeneously distributed in the mammalian brain including humans. In the mouse, only a few reports have assessed the expression of the MT1 melatonin receptor expression using 2-iodomelatonin binding, in situ hybridization and/or polymerase chain reaction (PCR). Here, we described a transgenic mouse in which red fluorescence protein (RFP) is expressed under the control of the endogenous MT1 promoter, by inserting RFP cDNA at the start codon of MTNR1a gene within a bacterial artificial chromosome (BAC) and expressing this construct as a transgene. The expression of RFP in the brain of this mouse was examined either directly under a fluorescent microscope or immunohistochemically using an antibody against RFP (RFP-MT1). RFP-MT1 expression was observed in many brain regions including the subcommissural organ, parts of the ependyma lining the lateral and third ventricles, the aqueduct, the hippocampus, the cerebellum, the pars tuberalis, the habenula and the habenula commissure. This RFP-MT1 transgenic model provides a unique tool for studying the distribution of the MT1 receptor in the brain of mice, its cell-specific expression and its function in vivo.
机译:松果体褪黑激素激活两个G蛋白偶联受体(MT1和MT2),以部分调节生物学功能。 MT1和MT2褪黑激素受体异质分布在包括人在内的哺乳动物大脑中。在小鼠中,只有很少的报道使用2-碘降钙素结合,原位杂交和/或聚合酶链反应(PCR)评估MT1褪黑素受体表达的表达。在这里,我们描述了一种转基因小鼠,其中在内源性MT1启动子的控制下表达红色荧光蛋白(RFP),方法是将RFP cDNA插入细菌人工染色体(BAC)内MTNR1a基因的起始密码子,并以此形式表达转基因。直接在荧光显微镜下或使用针对RFP的抗体(RFP-MT1)免疫组化检查该小鼠脑中RFP的表达。 RFP-MT1表达在许多脑区中观察到,包括连合下器官,侧脑室和第三脑室的室管膜部分,输水管,海马体,小脑,肺结节,the管和the管连合。此RFP-MT1转基因模型为研究MT1受体在小鼠脑中的分布,其细胞特异性表达及其体内功能提供了独特的工具。

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