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.

在人类中，就像在果蝇果蝇中一样，几种磷酸二酯酶（PDES）出口。他们水解营地以放大器并终止营地信号传导。 Houslay在2010年已经提出了与磷酸二酯酶功能相关的最重要问题之一：“为什么当所有这些都水解时会产生这么多不同的同工型？”虽然有新的证据表明，由于针对性的PDE活性和cAMP效应蛋白对细胞中不同领域而导致的cAMP信号特异性涉及cAMP的亚细胞微域，但我们的知识远非全面。我们建议使用果蝇Melanogaster在神经元和行为水平上使用PDE4D矫形器对亚细胞的功能。我们想研究是否需要在神经元中建立不同的cAMP微型域，是否需要使用PDE4D直系同源邓切的不同同工型。由于服务原因，将我们的分析重点放在邓斯上。第一个邓肯（First Dunce）是果蝇中唯一的cAMP特异性PDE，其次我们先前已经表明，不同的邓ce同工型在不同的亚细胞隔室中表达。第三个邓ce突变体在神经元可塑性方面的缺陷有所不同。为了衡量神经元可塑性和功能的变化，我们将分析Dunce在关联厌恶嗅觉学习和记忆和神经元形态中的功能。特别是我们要使用行为性，细胞下降水平的差异来解决不同亚细胞域中同一神经元中不同的邓ce同工型是否起作用以调节cAMP信号传导。此外，我们要确定在不同神经元中相同形式的神经元可塑性中相同的同工型是否调节。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，或者这些变化是否在整个神经元上延伸。为此，我们将使用遗传，分子遗传学，想象力技术和行为分析。获得的结果将阐明果蝇细胞cAMP结构域的建立以及不同的邓氏同工型如何促进神经元可塑性的变化。