Regulating the cocaine-sensitive serotonin transporter

调节可卡因敏感的血清素转运蛋白

• 批准号: 7426957
• 负责人: BARRY G CONDRON
• 金额: $ 20.88万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7426957
• 关键词: Attenuated; Cocaine; Complex; Data; Development; Eagles; Embryo; Embryonic Development; Equilibrium; Fibroblast Growth Factor; Fibroblast Growth Factor Receptors; Genes; Goals; Human; Larva; Mediating; Mental disorders; Methods; Modeling; Mutation; Neuraxis; Numbers; Pathway interactions; Phenotype; Protein Tyrosine Kinase; Regulation; Research; Role; Serotonin; Signal Pathway; Signal Transduction; Subcellular Anatomy; Synapses; Testing; Varicosity; Work; axon guidance; clinically relevant; early onset; extracellular; fly; member; mutant; research study; serotonin transporter; synaptogenesis; time use; transcription factor

DESCRIPTION (provided by applicant): The expression of the cocaine-sensitive serotonin transporter (SerT) is regulated in a complex manner in the developing central nervous system (CNS). Mutations that alter expression of SerT are associated with a number of human mental illnesses. Here we describe a set of experiments in the relatively simple fly CNS that test the role of certain genes in regulating SerT. In addition, we also propose to examine the effects of altering SerT expression on serotonergic cellular anatomy. Our previous experiments showed that the axon guidance molecule robo2 regulates SerT expression via the transcription factor eagle (eg). Here we test whether the cytoplasmic tyrosine kinase abl mediates robo2 signaling to eg. Our preliminary data indicate that members of the FGF signaling pathway function to inhibit SerT activity. In FGF, FGF-receptor, pointed (pnt) andros mutants, precocious SerT activity appears during embryonic development. Here we propose to test where and when these molecules function to attenuate SerT activity. Our working model is that the activation of embryonic SerT expression is a balance between robo2/eg induction and FGF/pnt inhibition. This balance is further revealed by the fact that the phenotype of each single mutant is suppressed in a pnt/eg double mutant. We therefore propose to examine how the balance of robo2 and FGF signaling regulates SerT. While the robo2/eg and FGF/pnt pathways seem to function during initial SerT onset early in development we will also test whether these same molecules continue to function as regulators in the CNS of the behaving larva. For this we will use timed expression of components of these pathways to test for later function. Extracellular serotonin levels, as regulated by SerT, have been hypothesized to regulate serotonergic synapse formation. We have developed a method to quantify serotonergic synaptic varicosities in the CNS. We will use this approach to first test the direct effects of altering SerT and serotonin levels on varicosities. We will then test the effects of altering genes that regulate SerT and serotonin. The long-term goal of this research is to further understand the regulation and function of this clinically relevant molecule.

描述（由申请人提供）：可卡因敏感的5-羟色胺转运蛋白（SERT）的表达在发育中的中枢神经系统（CNS）中以复杂的方式调节。改变SERT表达的突变与许多人类精神疾病有关。在这里，我们描述了一组相对简单的蝇中枢神经系统中测试某些基因在调节SERT中的作用的实验。此外，我们还建议研究改变SERT表达对5-羟色胺能细胞解剖结构的影响。我们以前的实验表明，轴突引导分子ROBO2通过转录因子鹰（EG）调节SERT表达。在这里，我们测试细胞质酪氨酸激酶ABL是否将Robo2信号介导为Eg。我们的初步数据表明，FGF信号通路途径功能的成员可以抑制SERT活性。在FGF中，FGF受体，尖（PNT）Andros突变体，在胚胎发育过程中出现早熟的SERT活性。在这里，我们建议测试这些分子在何处以及何时衰减SERT活性。我们的工作模型是，胚胎SERT表达的激活是ROBO2/EG诱导与FGF/PNT抑制之间的平衡。通过以下事实，每个单个突变体的表型在PNT/EG双重突变体中被抑制。因此，我们建议研究Robo2和FGF信号的平衡如何调节SERT。尽管ROBO2/EG和FGF/PNT途径在开发初期的初始SERT发作过程中似乎在起作用，但我们还将测试这些相同的分子是否继续充当行为幼虫中枢神经系统的调节剂。为此，我们将使用这些途径的组件的定时表达来测试以后的功能。已假设细胞外5-羟色胺水平受SERT的调节以调节5-羟色氨酸突触的形成。我们开发了一种量化CNS中血清素能突触静脉曲张的方法。我们将使用这种方法首先测试改变SERT和5-羟色胺水平对静脉曲张的直接影响。然后，我们将测试改变调节SERT和5-羟色胺的基因的影响。这项研究的长期目标是进一步了解该临床相关分子的调节和功能。