Expression patterns of promoters for NPYY1 and Y5 receptors in Y5RitTA and Y1RVenus BAC-transgenic mice
Expression patterns of promoters for NPYY1 and Y5 receptors in Y5RitTA and Y1RVenus BAC-transgenic mice
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DOI：
10.1111/j.1460-9568.2007.05631.x
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发表时间：
2007-07-01
影响因子：
3.4
通讯作者：
Eva, Carola
中科院分区：
文献类型：
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作者：
Oberto, Alessandra;Acquadro, Elena;Eva, Carola
在大鼠脑中，神经肽Y（NPY）Y-1和Y-5受体在不同的前脑区域共表达，在那里它们介导几种NPY激活的功能，包括进食行为、焦虑、神经元兴奋性和激素分泌。我们研究了Y-1和Y-5受体基因表达在小鼠脑中的分布模式和细胞共定位，通过使用基因组整合BAC克隆的转基因小鼠，其中Y-1和Y-5受体基因的编码区分别被Venus和合成转录因子itTA报告基因取代（Tg（Y 5 RitTA/Y1 RVenus）小鼠）。金星荧光和ITTA介导的激活Cre重组酶的分析揭示了拷贝数依赖的表达水平，线之间，但相似的表达模式。在三个转基因株系中，BAC编码的Y-5受体启动子诱导Cre在嗅觉系统、大脑皮层、海马和基底神经节中强烈表达。在表达最高的转基因品系25的大多数下丘脑核中发现较弱的表达。Cre的激活是itTA依赖性的，并且可以被多西环素调节。Y-1受体启动子诱导的金星荧光是强烈的，广泛存在于整个大脑和共定位与Cre免疫染色在不同的大脑区域，包括大脑皮层，基底外侧杏仁核，齿状回和室旁核的神经元。这些数据提供了小鼠脑细胞内Y-1和Y-5受体启动子活性的详细和比较图谱。这里产生的Tg（Y 5 RitTA/Y1 RVenus）转基因小鼠也代表了通过Cre lox系统进行条件诱变的遗传工具，特别是涉及摄食行为、神经元兴奋性和激素分泌的基因。
In the rat brain, neuropeptide Y (NPY) Y-1 and Y-5 receptors are coexpressed in various forebrain regions where they mediate several NPY-activated functions, including feeding behaviour, anxiety, neuronal excitability and hormone secretion. We studied the distribution pattern and cellular colocalization of the Y-1 and the Y-5 receptor gene expression in the mouse brain by using transgenic mice with genomically integrated BAC clones, where the coding regions of the Y-1 and Y-5 receptor genes were replaced by Venus and the synthetic transcription factor itTA reporter genes, respectively (Tg(Y5RitTA/Y1RVenus) mice). Analysis of Venus fluorescence and itTA-mediated activation of Cre recombinase revealed copy number-dependent expression levels, between the lines, but similar expression patterns. In three transgenic lines the BAC encoded Y-5 receptor promoter induced strong Cre expression in the olfactory system, cerebral cortex, hippocampus and basal ganglia. Weaker expression was found in most of the hypothalamic nuclei of line 25, the highest-expressing transgenic line. Activation of Cre was itTA-dependent and could be regulated by doxycycline. The Y-1 receptor promoter-induced Venus fluorescence was intense, widely present through the brain and colocalized with Cre immunostaining in neurons of distinct brain regions, including the cerebral cortex, basolateral amygdala, dentate gyrus and paraventricular nucleus. These data provide a detailed and comparative mapping of Y-1 and Y-5 receptor promoter activity within cells of the mouse brain. The Tg(Y5RitTA/Y1RVenus)-transgenic mice generated here also represent a genetic tool for conditional mutagenesis via the Cre lox system, particularly of genes involved in feeding behaviour, neuronal excitability and hormone secretion.