Gelman Diego M, Noaín Daniela, Avale M Elena, Otero Verónica, Low Malcolm J, Rubinstein Marcelo
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Consejo Nacional de Investigaciones Científicas y Técnicas and Departmento de Fisiología, Biología Molecular y Celular, Universidad de Buenos Aires, Argentina.
Genesis. 2003 Aug;36(4):196-202. doi: 10.1002/gene.10217.
To introduce restricted DNA recombination events into catecholaminergic neurons using the Cre/loxP technology, we generated transgenic mice carrying the Cre recombinase gene driven by a 9 kb rat tyrosine hydroxylase (TH) promoter. Immunohistochemistry performed on transgenic mouse brain sections revealed a high number of cells expressing Cre in areas where TH is normally expressed, including the olfactory bulb, hypothalamic and midbrain dopaminergic neurons, and the locus coeruleus. Double immunohistochemistry and immunofluorescence indicated that colocalization of TH and Cre is greater than 80%. Cre expression was also found in TH-positive amacrine neurons of the retina, chromaffin cells of the adrenal medulla, and sympathetic ganglia. We crossbred TH-Cre mice with the floxed reporter strain Z/AP and observed efficient Cre-mediated recombination in all areas expressing TH, indicating that transgenic Cre is functional. Therefore, we have generated a valuable transgenic mouse strain to induce specific mutations of "floxed" genes in catecholaminergic neurons.
为了利用Cre/loxP技术将受限的DNA重组事件引入儿茶酚胺能神经元，我们构建了携带由9 kb大鼠酪氨酸羟化酶（TH）启动子驱动的Cre重组酶基因的转基因小鼠。对转基因小鼠脑切片进行的免疫组织化学分析显示，在TH正常表达的区域有大量表达Cre的细胞，包括嗅球、下丘脑和中脑多巴胺能神经元以及蓝斑。双重免疫组织化学和免疫荧光表明，TH和Cre的共定位率大于80%。在视网膜的TH阳性无长突细胞、肾上腺髓质的嗜铬细胞和交感神经节中也发现了Cre表达。我们将TH-Cre小鼠与floxed报告菌株Z/AP进行杂交，发现在所有表达TH的区域均发生了高效的Cre介导的重组，表明转基因Cre具有功能。因此，我们构建了一种有价值的转基因小鼠品系，可用于在儿茶酚胺能神经元中诱导“floxed”基因的特异性突变。
