Nausch Norman, Manteuffel Gerhard, Vanselow Jens
Division of Innate Immunity, German Cancer Research Center, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
J Steroid Biochem Mol Biol. 2007 Feb;103(2):119-28. doi: 10.1016/j.jsbmb.2006.08.007. Epub 2006 Oct 31.
Cyp19 encodes the key enzyme of estrogen biosynthesis, aromatase cytochrome P450. In mice it is mainly expressed in the ovary and brain, where transcription is directed by a distal, brain-specific promoter (P(br)). In order to map functional sequence elements of P(br), portions of various length (0.2, 1.0, and 1.7[kb]) were fused to a lacZ reporter gene and analyzed in transgenic mice. Numbers of integrated reporter genes varied from 1 to 23 copies in different transgenic lines. These copy numbers however did not show any correlation to the levels of transgene expression. All of the constructs were found being expressed in the olfactory bulb, limbic cortex, amygdala, and hypothalamus. Additional expression in thalamic nuclei, bed nucleus of stria terminalis, and dorsal mesencephalon was found in transgenic lines with constructs 1.0 and 1.7, and expression in septal and preoptic nuclei was only found with construct 1.7. The data demonstrate that 0.2kb of P(br) target reporter gene expression to specific brain areas. The data also strongly suggest that the sequence between 0.2 and 1.7kb upstream, is necessary for expression in additional areas. However even 1.7kb of P(br) are not sufficient to consistently mimic the accurate expression pattern of Cyp19.
Cyp19编码雌激素生物合成的关键酶——芳香化酶细胞色素P450。在小鼠中,它主要在卵巢和大脑中表达,其转录由一个远端的、大脑特异性启动子(P(br))指导。为了绘制P(br)的功能序列元件图谱,将不同长度(0.2、1.0和1.7[kb])的片段与lacZ报告基因融合,并在转基因小鼠中进行分析。在不同的转基因品系中,整合的报告基因数量从1到23个不等。然而,这些拷贝数与转基因表达水平没有任何相关性。所有构建体均在嗅球、边缘皮质、杏仁核和下丘脑表达。在含有1.0和1.7构建体的转基因品系中,在丘脑核、终纹床核和中脑背侧发现了额外的表达,而仅在含有1.7构建体的品系中,在隔核和视前核发现了表达。数据表明,0.2kb的P(br)将报告基因的表达靶向特定脑区。数据还强烈表明,上游0.2至1.7kb之间的序列对于在其他区域的表达是必需的。然而,即使1.7kb的P(br)也不足以始终如一地模拟Cyp19的准确表达模式。