Cocaine Regulation of Norepinephrine Transporter

可卡因对去甲肾上腺素转运蛋白的调节

• 批准号: 6928419
• 负责人: LANKUPALLE D JAYANTHI
• 金额: $ 7.3万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6928419
• 关键词: biological signal transduction; cocaine; enzyme induction /repression; laboratory rat; membrane proteins; messenger RNA; neurotransmitter metabolism; neurotransmitter receptor; neurotransmitter transport; norepinephrine; northern blottings; polymerase chain reaction; posttranslational modifications; protein kinase; receptor expression; tissue /cell culture; trophoblast; western blottings

DESCRIPTION (provided by applicant): The catecholamine, norepinephrine (NE) governs various physiologic processes from vasoconstriction and heart rate to attention and motivation. NE signaling is tightly controlled by a diverse set of macromolecules including biosynthetic enzymes, secretory proteins, ion channels, pre- and post synaptic receptors and NE transporters (NETs). NE signaling is terminated primarily by active reuptake of the catecholamine via cocaine- and amphetamine-sensitive norepinephrine transporters (NETs). Various biologic stimuli are known to regulate NE signaling, and alterations in NE signaling including NE clearance and NET density are observed in cardiovascular diseases and brain disorders. The triggers and molecular mechanisms of transporter regulation are important in the control of extracellular NE concentrations and hence NE signaling. NETs are also expressed in the placenta. This raises the possibility that the established medical complications associated with the maternal use of psychostimulant drugs may arise in part from blocking placental NET. Primary cell cultures offer the facility of in vitro experimentation combined with the verisimilitude of a native cell system to study the molecular mechanisms of transporter regulation a step closer to in vivo animal models. To explore the effect of psychostimulants on NET regulatory pathways, we have developed primary cultures of placental trophoblasts that express endogenous NETs. Using trophoblast cell cultures, we show that NET function and expression are regulated by 1) cocaine treatment, 2) receptor modulation, and 3) kinase(s) activation. Together, these preliminary findings support the proposed experiments to test a specific hypothesis that cocaine regulates NET function and expression via altered cellular mechanisms that result from cocaine effects on NET and/or other cellular targets. The studies proposed in this application will identify the mechanisms of cocaine's action in regulating the function and expression of native NET. Results from these studies will provide valuable scientific insights to our understanding of the role of drugs of abuse in regulating NET function and expression.

描述（由申请人提供）：儿茶酚胺、去甲肾上腺素（NE）控制着从血管收缩和心率到注意力和动机的各种生理过程。 NE 信号传导受到多种大分子的严格控制，包括生物合成酶、分泌蛋白、离子通道、突触前和突触后受体以及 NE 转运蛋白 (NET)。 NE 信号传导主要是通过可卡因和安非他明敏感的去甲肾上腺素转运蛋白 (NET) 主动再摄取儿茶酚胺来终止的。已知多种生物刺激可调节 NE 信号传导，并且在心血管疾病和脑部疾病中观察到 NE 信号传导的改变，包括 NE 清除率和 NET 密度。转运蛋白调节的触发因素和分子机制对于细胞外 NE 浓度以及 NE 信号传导的控制非常重要。 NETs 也在胎盘中表达。这提出了一种可能性，即与母亲使用精神兴奋药物相关的已确定的医疗并发症可能部分是由于阻断胎盘 NET 引起的。原代细胞培养提供了体外实验的便利性以及与天然细胞系统的逼真性相结合，以研究转运蛋白调节的分子机制，更接近于体内动物模型。为了探索精神兴奋剂对 NET 调节途径的影响，我们开发了表达内源性 NET 的胎盘滋养层原代培养物。使用滋养层细胞培养物，我们发现 NET 功能和表达受 1) 可卡因治疗、2) 受体调节和 3) 激酶激活的调节。总之，这些初步研究结果支持了所提出的实验，以测试可卡因通过改变细胞机制来调节 NET 功能和表达的特定假设，而细胞机制是可卡因对 NET 和/或其他细胞靶标的影响所导致的。本申请中提出的研究将确定可卡因调节天然 NET 功能和表达的作用机制。这些研究的结果将为我们理解滥用药物在调节 NET 功能和表达中的作用提供有价值的科学见解。