Functional coding variation in recombinant inbred mouse lines reveals multiple serotonin transporter-associated phenotypes
Functional coding variation in recombinant inbred mouse lines reveals multiple serotonin transporter-associated phenotypes
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DOI：
10.1073/pnas.0809449106
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发表时间：
2009-02-10
影响因子：
11.1
通讯作者：
Blakely, Randy D.
中科院分区：
文献类型：
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作者：
Carneiro, Ana M. D.;Airey, David C.;Blakely, Randy D.
人血清素（5-羟色胺，5-HT）转运体（hSERT，SLC 6A 4）在许多流行的神经行为障碍（包括焦虑、酒精中毒、抑郁、自闭症和强迫症（OCD））的病因学和治疗中占有突出地位。在这里，我们使用重组近交系（RI）中自然发生的多态性来鉴定与SERT功能改变相关的多种表型。广泛使用的小鼠品系C57 BL/6 J具有由2个非同义编码变体[Gly-39和Lys-152（GK）]定义的SERT单倍型。在这些位置，许多其他小鼠品系，包括DBA/2 J，分别编码Glu-39和Arg-152（ER单倍型），这些氨基酸也在hSERT中发现。离体突触体5- HT转运研究显示与GK变体相关的摄取减少，这一发现得到了体外异源表达研究的证实。使用RI系（C57 BL/6 J x DBA/2 J = BXD）的实验和计算机模拟方法鉴定了多种特别受GK/ER变异影响的解剖、生化和行为表型。在我们的研究结果中，有几个与饮酒相关的特征和多个与多巴胺信号相关的特征。BXD表型的进一步生物信息学分析，结合SERT基因敲除小鼠的生化评价，提名SERT依赖性5-HT信号传导作为中脑铁稳态的主要决定因素，这反过来又决定了铁调节的DA表型。我们的研究提供了一个例子，协调在体外，在体内，并在硅片方法使用小鼠RI线，以阐明和量化基因变异的系统水平的影响。
The human serotonin (5-hydroxytryptamine, 5-HT) transporter (hSERT, SLC6A4) figures prominently in the etiology and treatment of many prevalent neurobehavioral disorders including anxiety, alcoholism, depression, autism, and obsessive-compulsive disorder (OCD). Here, we use naturally occurring polymorphisms in recombinant inbred ( RI) lines to identify multiple phenotypes associated with altered SERT function. The widely used mouse strain C57BL/6J, harbors a SERT haplotype defined by 2 nonsynonymous coding variants [Gly-39 and Lys-152 (GK)]. At these positions, many other mouse lines, including DBA/2J, encode, respectively, Glu-39 and Arg-152 ( ER haplotype), amino acids found also in hSERT. Ex vivo synaptosomal 5- HT transport studies revealed reduced uptake associated with the GK variant, a finding confirmed by in vitro heterologous expression studies. Experimental and in silico approaches using RI lines (C57BL/6J x DBA/2J = BXD) identify multiple anatomical, biochemical, and behavioral phenotypes specifically impacted by GK/ER variation. Among our findings are several traits associated with alcohol consumption and multiple traits associated with dopamine signaling. Further bioinformatic analysis of BXD phenotypes, combined with biochemical evaluation of SERT knockout mice, nominates SERT-dependent 5-HT signaling as a major determinant of midbrain iron homeostasis that, in turn, dictates iron-regulated DA phenotypes. Our studies provide an example of the power of coordinated in vitro, in vivo, and in silico approaches using mouse RI lines to elucidate and quantify the system-level impact of gene variation.