Circuitry and Modulation of the Circardian Clock Network of D. melanogaster

在动物中，睡眠/活动节律所必需的计时发生在位于离散神经元中的神经元中。 中枢神经系统（CNS）的区域。值得注意的是，个体神经元生物钟可以维持 在没有来自环境或其他细胞的时间线索的情况下的分子和生理节律。尽管 这种细胞自主计时，日常行为节奏的协调依赖于时钟网络， 神经元我试图了解这些网络的电路特性，并检查确定的角色， 神经元在控制昼夜节律中发挥作用。在我的研究的ROO阶段！将使用解剖学， 在我的资助的K99阶段，我开发了基因和实时成像技术，以发现时间是如何保持的 以及它是如何用来协调果蝇的日常和季节性行为变化的 黑腹菌此外，新开发的活体成像技术、苍蝇遗传学和光线控制技术， 神经元信号将被开发来研究果蝇生物钟网络的电路特性 个脑袋了解神经元时钟网络的组织，以及 神经系统中的神经调节信号，我提出了以下具体目标：1）开发 实时成像方法，用于同时操纵和观察神经元信号， 细胞兴奋性的光遗传学控制与cAMP和Ca 2-f，H的遗传编码传感器结合 神经元时钟网络的肽/胺调节剂的鉴定。三）识别和 使用实时成像和靶向遗传标记表征时钟神经元输出的非时钟目标 IV）这些神经元内的GPCR/cAMP/Ca 2-t-信号传导的遗传解剖。的 我的K99资助支持的工作支持了拟议研究的各个方面的可行性， ROO提供的资源将使我能够在研究中使用先进的成像技术， 神经时钟网络此外，这里提出的工作将涉及以下基本方面： 神经生物学和创造新的方法来研究神经元回路和控制动物 行为

In animals the timekeeping necessary for sleep/activity rhythms takes place within neurons located in discrete regions of the central nervous system (CNS). Remarkably, individual neuronal circadian clocks can maintain molecular and physiological rhythms in the absence of time-cues from the environment or other cells. Despite such cell-autonomous timekeeping, the orchestration of daily behavioral rhytiims depends on networks of clock neurons. I seek to understand the circuit properties of these networks and to examine the roles that identified neurons play in the control of circadian rhythms. During the ROO phase of my research! will use the anatomical, genetic, and live-imaging techniques 1 developed during the K99 phase of my grant to discover how time is kept within the brain and how it is used to orchestrate daily and seasonal changes in behavior in the fly Drosophila melanogaster. Furthermore, newly developed technologies in live imaging, fly genetics, and light-control of neuronal signaling will be developed to investigate the circuit properties of the circadian clock network in the fly brain. To understand the organization of the neuronal clock network, and fundamental questions of neuromodulatory signaling in the nervous system, I propose the following speciflc aims: 1) The development of live Imaging methods for the simultaneous manipulation and observation of neuronal signaling using optogenetic control of cell excitability in conjunction with genetically encoded sensors for cAMP and Ca2-f, H) The identiflcation of peptide/amine modulators of the neuronal clock network. Ill) The identiflcation and characterization of non-clock targets of clock neuron output using live imaging and targeted genetic manipulation approaches, IV) The genetic dissection of GPCR/cAMP/Ca2-t- signaling within these neurons. The work supported by my K99 grant supports the feasibility of all aspects of the proposed research and the resources made possible by the ROO will allow me to employ advanced imaging technologies in my study of the neuronal clock network. Furthermore, the work proposed here will address fundamental aspects of neurobiology and create new methodologies for the investigation of neuronal circuitry and the control of animal behavior.