Project 3 Signaling Networks Sustaining Serotonin Transport

突触前5-羟色胺转运体(SERT)控制释放后5-羟色胺的可用性，并回收5-羟色胺 在随后的释放事件中重复使用，从而有助于突触前5HT的动态平衡。Sserts是 SERT中最常用的抗抑郁药物和基因变异的靶点 与自闭症、焦虑症、严重抑郁障碍(MOD)和抗抑郁药物反应有关。镜像 依赖有效的5-羟色胺清除来决定5-羟色胺反应的突触后基因网络 相互作用的蛋白质和细胞信号通路的突触前网络决定适当的SERT表面 丰度和催化活性。布莱克利实验室一直是SERT分子领域的领先者 自首次在小鼠和小鼠中发现SERT基因以来十多年的生物学和调控 天哪。在项目3：支持5-羟色胺运输的信号网络中，布莱克利的团队提出了三个 目的是更全面地阐明SERT调节蛋白质组在体内的识别和调控，提供 新的靶点和模型，以丰富我们对5HT信号如何建立和调制的理解。在……里面 具体目标I，Blakely将使用候选和蛋白质组方法来阐明SERT调控 在血小板中建立的网络，是SERT在外围的丰富来源，其次是生化和 对神经元中共表达的解剖学验证，以及对网络稳定性的评估 SERT调节激酶和磷酸酶的激活。在《特定目标2》中，布莱克利提出了创造 以及限制PKG介导的SERT调节的转基因小鼠模型的评估，或者通过 SERT中PKG1的结构性和RAPH特异性丢失或关键的PKG磷酸化位点的消除。 其次，布莱克利评估了5-羟色胺和抗抑郁剂对SERT运输和蛋白质的活性 通过研究携带降低SSRI和可卡因识别能力的lle172Met等位基因的小鼠的相关性 在SERT时不丧失5-羟色胺摄取功能。最终目标是将在SERT活动中观察到的变化联系起来 在这些模型中对Aim I中确定的SERT蛋白质组的稳定性和组织进行了研究 BLJIlakely的团队将研究工程突变的更广泛的生理影响，与 ONTE调查人员探讨其对体内5-羟色胺动态平衡和SERT活性的影响 5-羟色胺受体的IF‘处理(编辑)和信号传递，以及生理和行为效应的监测 通过透析和计时安培研究以及既定的行为技术。一起， 这些努力将更广泛地阐明SERT调控和SERT调控蛋白质组是如何 在体内建立适当的5-羟色胺清除能力以及遗传变异如何影响SERT调节。

Presynaptic 5HT transporters (SERT) control the availability of 5HT following release and recycle 5HT for reuse in subsequent release events, thereby contributing to presynaptic 5HT homeostasis. SERTs are targets for the most commonly prescribed antidepressant medications and genetic variation in SERT has been linked to autism, anxiety, major depressive disorder (MOD) and antidepressant response. Mirroring the network of postsynaptic genes that depend on efficient 5HT clearance to dictate 5HT response, a presynaptic network of interacting proteins and cell signaling pathways dictates appropriate SERT surface abundance and catalytic activity. The Blakely laboratory has been a leader in the field of SERT molecular biology and regulation for over a decade originating with the first identification of SERT genes in mouse and man. In Project 3: Signaling Networks Supporting Serotonin Transport, Blakely's team proposes three Aims to more fully elucidate the identity and regulation of the SERT regulatory proteome in vivo, providing new targets and models to enrich our understanding of how 5HT signaling is established and modulated. In Specific Aim I, Blakely will use both candidate and proteomic approaches to illuminate the SERT regulatory network established in platelets, a rich source for SERT in the periphery, followed by biochemical and anatomical validation of co-expression in neurons, and assessment of the stability of the network to activation of SERT regulatory kinases and phosphatases. In Specific Aim II, Blakely proposes the creation and evaluation of transgenic mouse models that limit PKG mediated regulation of SERT, either through constitutive and raph-specific loss of PKG1 or elimination of a key PKG phosphorylation site in SERT. Secondly, Blakely evaluates the activity of 5HT and antidepressants on SERT trafficking and protein associations via studies of mice harboring the lle172Met allele which reduces SSRI and cocaine recognition at SERT without loss of 5HT uptake function. The ultimate goal is to link changes observed in SERT activity in these models to the stability and organization of the SERT proteome identified in Aim I. In Specific Aim I BLJIlakely's team will examine the broader physiological impact of engineered mutations, collaborating with onte Investigators to explore their impact on 5HT homeostasis and SERT activity in vivo, the abundance If'recessing (editing) and signaling of 5HT receptors, and the physiological and behavioral effects, monitored through dialysis and chronoamperometry studies as well as established behavioral techniques. Together, these efforts will more broadly elucidate how SERT regulation and the SERT regulatory proteome establishes proper 5HT clearance capacity and how genetic variation can influence SERT regulation in vivo.