Gene expression in lungs of mice lacking the 5-hydroxytryptamine transporter gene.
Gene expression in lungs of mice lacking the 5-hydroxytryptamine transporter gene.
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DOI：
10.1186/1471-2466-9-19
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发表时间：
2009-05-10
影响因子：
3.1
通讯作者：
West, James
中科院分区：
文献类型：
--
作者：
Crona, Daniel;Harral, Julie;West, James
背景：近40年来，血清素转运体（5HTT）的调节已被证明是肺动脉高压的一个危险因素，但缺乏关于5HTT肺抑制的广泛分子效应的体内数据。先前的研究表明对炎症、增殖和血管收缩有影响。这项研究的目的是确定其中哪些是由血清素转运蛋白敲除小鼠基因表达的改变所支持的。方法：对5-HTT基因敲除（5HTT-/-）和杂合子（5HTT+/-）或野生型（5HTT+/+）的8周龄正常小鼠及其窝伴进行右心室收缩压（RVSP）评估，收集肺部RNA，汇总，并在Affymetrix阵列分析中重复使用。采用定量RT-PCR和western blot对代表性基因进行鉴定。结果：各组RVSP均正常。在5HTT-/-和5HTT+/+小鼠之间，只有124个基因发生了可靠的变化。其中一半以上与炎症反应或肌肉功能和组织有关;此外，一些基质、血红素加氧酶、发育和能量代谢基因的表达也发生了改变。每个主要组样本的定量RT-PCR证实了通过阵列观察到的变化，在5HTT +/-小鼠中具有中等水平。结论：这些结果首次显示了5HTT基因敲除在肺中的体内效应，并表明细胞培养和离体实验提出的许多下游机制在体内也可运作。这表明5HTT对肺血管功能的影响源于其对几个系统的影响，包括血管反应性、增殖和免疫功能。
BACKGROUND: While modulation of the serotonin transporter (5HTT) has shown to be a risk factor for pulmonary arterial hypertension for almost 40 years, there is a lack of in vivo data about the broad molecular effects of pulmonary inhibition of 5HTT. Previous studies have suggested effects on inflammation, proliferation, and vasoconstriction. The goal of this study was to determine which of these were supported by alterations in gene expression in serotonin transporter knockout mice.METHODS: Eight week old normoxic mice with a 5-HTT knock-out (5HTT-/-) and their heterozygote(5HTT+/-) or wild-type(5HTT+/+) littermates had right ventricular systolic pressure(RVSP) assessed, lungs collected for RNA, pooled, and used in duplicate in Affymetrix array analysis. Representative genes were confirmed by quantitative RT-PCR and western blot.RESULTS: RVSP was normal in all groups. Only 124 genes were reliably changed between 5HTT-/- and 5HTT+/+ mice. More than half of these were either involved in inflammatory response or muscle function and organization; in addition, some matrix, heme oxygenase, developmental, and energy metabolism genes showed altered expression. Quantitative RT-PCR for examples from each major group confirmed changes seen by array, with an intermediate level in 5HTT +/- mice.CONCLUSION: These results for the first time show the in vivo effects of 5HTT knockout in lungs, and show that many of the downstream mechanisms suggested by cell culture and ex vivo experiments are also operational in vivo. This suggests that the effect of 5HTT on pulmonary vascular function arises from its impact on several systems, including vasoreactivity, proliferation, and immune function.