Dissecting the PDE4d orthologue Dunce isoform functions in Drosophila melanogaster

剖析果蝇中 PDE4d 直系同源 Dunce 亚型功能

• 批准号: 407177148
• 负责人: Professorin Dr. Henrike Scholz
• 金额: --
• 依托单位: AG Molecular Mechanisms of Addcitive Behaviors
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/407177148?language=en
• 关键词: Dissecting; PDE4d; orthologue; Dunce; isoform

In humans as in the fruit fly Drosophila melanogaster several phosphodiesterases (PDEs) exit. They hydrolyze cAMP to AMP and terminate cAMP signaling. One of the most important questions related to phosphodiesterase function was already raised by Houslay in 2010: “Why are there so many different isoforms generated when all of them hydrolyze cAMP?” While there is emerging evidence that sub-cellular micro-domains of cAMP are involved in the regulation of cAMP signaling specificity due to targeted PDE activity and cAMP effector proteins to different domains in the cell, our knowledge is far from comprehensive. We propose to use Drosophila melanogaster to diessect the function of the Pde4d orthologue Dunce on subcellular, on neuronal and behavioral level. We want to investigate whether different isoforms of the Pde4D orthologue Dunce are required to set up distinct cAMP micro-domains in neurons. We focus our analysis on Dunce for serval reasons. First Dunce is the only cAMP specific PDE in Drosophila, second we have previosly shown that different Dunce isoforms are expressed in different sub-cellular compartments. Third dunce mutants differ in their defects in neuronal plasticity. To measure changes in neuronal plasticity and function we will analyze the function of Dunce in associative aversive olfactory learning and memory and neuron morphology. In particular we want to use the differences on behavioral, cellular und subcellular level to address whether different Dunce isoforms act in the same neurons in different subcellular domains to regulate cAMP signaling. Further we want to determine whether the same isoform regulates in different neurons similar forms of neuronal plasticity. Finally we will investigate whether subcellular changes of PDE activity are restricted to specific cellular sites or whether these changes extend over the whole neuron. Towards this end, we will use genetic, molecular-genetic, imagine techniques and behavioral analysis. The obtained results will shed light on how in Drosophila cellular cAMP domains are set up and how different Dunce isoforms contribute to changes of neuronal plasticity.

人类体内和果蝇果蝇中都存在多种磷酸二酯酶 (PDE)。它们将 cAMP 水解为 AMP 并终止 cAMP 信号传导。 Houslay 在 2010 年就已经提出了与磷酸二酯酶功能相关的最重要的问题之一：“为什么当它们全部水解 cAMP 时会产生如此多不同的亚型？”虽然有新的证据表明，由于 PDE 活性和 cAMP 效应蛋白针对细胞中不同的结构域，cAMP 的亚细胞微结构域参与 cAMP 信号传导特异性的调节，但我们的知识还远远不够全面。我们建议使用果蝇来剖析 Pde4d 直系同源物 Dunce 在亚细胞、神经元和行为水平上的功能。我们想要研究 Pde4D 直向同源基因 Dunce 的不同亚型是否需要在神经元中建立不同的 cAMP 微结构域。出于以下几个原因，我们将分析重点放在 Dunce 身上。首先Dunce是果蝇中唯一的cAMP特异性PDE，其次我们之前已经证明不同的Dunce亚型在不同的亚细胞区室中表达。第三个笨蛋突变体的不同之处在于神经元可塑性的缺陷。为了测量神经元可塑性和功能的变化，我们将分析 Dunce 在联想厌恶嗅觉学习和记忆以及神经元形态中的功能。特别是，我们希望利用行为、细胞和亚细胞水平上的差异来解决不同 Dunce 亚型是否作用于不同亚细胞域的相同神经元来调节 cAMP 信号传导。此外，我们想确定相同的亚型是否在不同神经元中调节相似形式的神经元可塑性。最后，我们将研究 PDE 活性的亚细胞变化是否仅限于特定的细胞位点，或者这些变化是否扩展到整个神经元。为此，我们将使用遗传学、分子遗传学、想象技术和行为分析。获得的结果将揭示果蝇细胞 cAMP 结构域的设置方式以及不同 Dunce 亚型如何影响神经元可塑性的变化。