Bäckman Cristina M, Malik Nasir, Zhang Yajun, Shan Lufei, Grinberg Alex, Hoffer Barry J, Westphal Heiner, Tomac Andreas C
Cellular Neurobiology Branch, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD 21224, USA.
Genesis. 2006 Aug;44(8):383-90. doi: 10.1002/dvg.20228.
Dopamine (DA) neurotransmission has been implicated in several neurological and psychiatric disorders. The dopamine transporter (DAT) is highly expressed in dopaminergic neurons of the ventral mesencephalon and regulates neurotransmission by transporting DA back into the presynaptic terminals. To mediate restricted DNA recombination events into DA neurons using the Cre/loxP technology, we have generated a knockin mouse expressing Cre recombinase under the transcriptional control of the endogenous DAT promoter. To minimize interference with DAT function by preservation of both DAT alleles, Cre recombinase expression was driven from the 3' untranslated region (3'UTR) of the endogenous DAT gene by means of an internal ribosomal entry sequence. Crossing this murine line with a LacZ reporter showed colocalization of DAT immunocytochemistry and beta-galactosidase staining in all regions analyzed. This knockin mouse can be used for generating tissue specific knockouts in mice carrying genes flanked by loxP sites, and will facilitate the analysis of gene function in dopaminergic neurons.
多巴胺（DA）神经传递与多种神经和精神疾病有关。多巴胺转运体（DAT）在腹侧中脑的多巴胺能神经元中高度表达，并通过将DA转运回突触前终末来调节神经传递。为了利用Cre/loxP技术介导限制的DNA重组事件进入DA神经元，我们构建了一种敲入小鼠，其在內源DAT启动子的转录控制下表达Cre重组酶。为了通过保留两个DAT等位基因来最小化对DAT功能的干扰，借助内部核糖体进入序列从內源DAT基因的3'非翻译区（3'UTR）驱动Cre重组酶表达。将该小鼠品系与LacZ报告基因杂交，结果显示在所有分析区域中DAT免疫细胞化学和β-半乳糖苷酶染色共定位。这种敲入小鼠可用于在携带loxP位点侧翼基因的小鼠中产生组织特异性敲除，并将有助于分析多巴胺能神经元中的基因功能。
