Identifying Novel Photic Inputs to the Drosophila Circadian/Arousal Neural Network for Behavioral Manipulation

项目摘要/摘要 蚊子病媒的控制主要依靠有毒杀虫剂。一种更环保的 另一种选择是利用基于光的行为操纵来吸引害虫到陷阱并击退害虫 从人类居住的地方。目前的技术是基于蚊子紫外线检测的假设 仅通过眼睛中基于视蛋白的光接收而发生。福尔摩斯实验室最近发现了更多 紫外光和短波长光感受元件在中枢神经元中的表达 昆虫对光的复杂行为反应。因此，有必要纳入这些额外的要素 在疾病媒介控制设计中提高效率。也就是说，隐色素(CRY)，这是经典的 与其在生物钟重置中的作用有关，在蓝光和紫外光下激活，并增加 昼夜节律/觉醒神经元的兴奋性。根据我的初步数据，我假设哭声与 其他光感受器介导光诱导的神经元电兴奋性，是复合体的基础 睡眠/清醒和昼夜节律调节了回避/吸引行为。 我的目标是确定哭声光传导的机制，我们如何操纵它 与其他光输入一起调节对光的复杂行为反应的机制，如果我们可以 将这些知识转化为蚊子系统，以进行基于光线的控制。我将从检查分子开始 大鼠昼夜节律/觉醒神经元电兴奋性的光传导机制 目标1。下一步，我将测量不同光感受器输入的相对贡献，它们介导了 Aim 2中昼夜节律/觉醒神经元的电生理光反应。最后，我将检查分子 表达昼夜叮咬的转基因果蝇的光信号转导机制及光反应 蚊子在AIM 3哭泣。我的研究将有助于开发针对特定物种的创新LED设备 在正在进行的防治媒介传播疾病的斗争中进行有害的昆虫控制。 我的赞助人托德·霍姆斯博士和我已经制定了一项培训计划，专注于发展我的 技术、写作和沟通能力，以及指导技能。我的技术技能的发展将重点放在 电生理学、行为分析和显微镜检查。我计划报名参加相关课程，参加 举办讲习班和培训活动，并与该领域的专家建立网络。我写作的发展和 沟通技能将通过申请助学金/奖学金、手稿开发和 在会议和座谈会上发表演讲。此外，我将通过培训来发展我的指导技能 帮助本科生进行实验和分析数据。我是一个高度合作的研究的一部分 环境，有许多电生理学、神经生物学和行为学、显微镜、遗传学和 我部门的生物信息学。我计划充分利用现有的资源和设施，以便 完成这个提议的目标，以及实现我成为一名独立研究科学家的目标。

PROJECT SUMMARY/ABSTRACT Mosquito disease vector control relies mostly on toxic insecticides. A more environmentally friendly alternative is to make use of light-based behavioral manipulation to attract pests to traps and repel pests away from human habitation. The present technology is based on the assumption that mosquito UV light detection occurs solely through opsin-based photoreception in the eyes. The Holmes Lab has recently found additional UV and short wavelength photoreceptive elements expressed in central brain neurons that strongly modulate complex insect behavioral responses to light. Therefore, there is a need to incorporate these additional elements in disease vector control designs for improved efficiency. Namely, CRYPTOCHROME (CRY), which is classically associated with its role in circadian clock resetting, activates with blue- and UV- light and increases the electrical excitability of circadian/arousal neurons. Based on my preliminary data, I hypothesize that CRY coordinates with other photoreceptors to mediate light-induced electrical excitability of neurons, which underlie complex sleep/wake and circadian modulated avoidance/attraction behaviors. My objectives are to determine the mechanism of CRY phototransduction, how we can manipulate that mechanism along with other light inputs to modulate complex behavioral responses to light, and if we can translate that knowledge to a mosquito system for light-based control. I will begin by examining the molecular phototransduction mechanism that underlie CRY-mediated electrical excitability of circadian/arousal neurons in Aim 1. Next, I will measure the relative contribution of different photoreceptor inputs that mediate the electrophysiological photoresponse in circadian/arousal neurons in Aim 2. Last, I will examine the molecular phototransduction mechanism and photic response of transgenic Drosophila expressing day-versus night-biting mosquito CRYs in Aim 3. My research will be useful for developing innovative LED devices for species-specific harmful insect control in the ongoing fight against vector-borne diseases. My Sponsor, Dr. Todd Holmes, and I have developed a training plan to focus on the development of my technical, writing and communication, and mentorship skills. Development of my technical skills will focus heavily on electrophysiology, behavioral assays, and microscopy. I plan to register for relevant courses, attend workshops and training events, and network with experts in the field. Development of my writing and communication skills will be accomplished by applying for grants/fellowship, manuscript development, and presenting at conferences and symposiums. Additionally, I will develop my mentorship skills by training undergraduate students to help run experiments and analyze data. I am a part of a highly collaborative research environment, with many experts in electrophysiology, neurobiology and behavior, microscopy, genetics, and bioinformatics within my department. I plan to fully utilize the resources and facilities available to me in order to accomplish the goals of this proposal, as well as achieve my goal of becoming an independent research scientist.