A Caenorhabditis elegans model to study dopamine transporter deficiency syndrome
A Caenorhabditis elegans model to study dopamine transporter deficiency syndrome
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DOI：
10.1111/ejn.13366
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发表时间：
2017-01-01
影响因子：
3.4
通讯作者：
Di Schiavi, Elia
中科院分区：
文献类型：
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作者：
Illiano, Placido;Lanzo, Ambra;Di Schiavi, Elia
多巴胺转运蛋白缺乏综合征（DTDS）是一种由多巴胺转运蛋白（DAT）基因突变引起的常染色体隐性遗传疾病。DTDS是一种耐药性综合征，其神经生物学知之甚少，部分原因是缺乏相关的动物模型。本研究的目的是建立第一个DTDS的动物模型，具有较强的结构效度，使用秀丽隐杆线虫，并调查人体DAT（hDAT）中发现的DTDS相关突变所发挥的体内作用。我们利用秀丽隐杆线虫敲除hDAT直向同源物cedat-1的优势，在空背景下获得携带野生型和两种突变形式（399 delG和941 C>T）的hDAT的遗传人源化动物。在表达人野生型形式的秀丽隐杆线虫转基因动物中，我们观察到敲除表型的拯救，如使用两种公认的范例所评估的，已知通过DAT对多巴胺或6-羟基多巴胺（6-OHDA）的内源性摄取来调节。不太严重的突变（941 C>T）能够部分拯救仅一种敲除表型，而399 delG突变在两种表型范例中都损害DAT功能。我们的体内表型研究结果表明，人类和线虫DAT之间的功能保守性，并验证了以前的体外指标的DTDS相关突变的载体中的hDAT的功能丧失。总而言之，这些观察结果使秀丽隐杆线虫成为一种新型动物模型，可在功能和体内测试中快速且廉价地筛选hDAT突变。
Dopamine transporter deficiency syndrome (DTDS) is a novel autosomal recessive disorder caused by mutations in the dopamine transporter (DAT), which leads to the partial or total loss of function of the protein. DTDS is a pharmacoresistant syndrome and very little is known about its neurobiology, in part due to the lack of relevant animal models. The objective of this study was to establish the first animal model for DTDS with strong construct validity, using Caenorhabditis elegans, and to investigate the invivo role played by DTDS-related mutations found in human DAT (hDAT). We took advantage of a C.elegans knockout for the hDAT orthologue, cedat-1, to obtain genetically humanized animals bearing hDAT, in the wild type and in two mutated forms (399delG and 941C>T), in a null background. In C.elegans transgenic animals expressing the human wild-type form, we observed a rescue of the knockout phenotype, as assessed using two well-established paradigms, known to be regulated by the endogenous uptake of dopamine or 6-hydroxydopamine (6-OHDA) by DAT. The less severe mutation (941C>T) was able to partially rescue only one of the knockout phenotypes, whereas the 399delG mutation impaired DAT function in both phenotypic paradigms. Our invivo phenotypic findings demonstrate a functional conservation between human and nematode DAT and validate previous invitro indications of the loss of function of hDAT in carriers of DTDS-related mutations. Taken together, these observations establish C.elegans as a novel animal model for fast and inexpensive screening of hDAT mutations in functional and invivo tests.