Neuronal basis of pheromone perception in mosquitoes

蚊子信息素感知的神经元基础

• 批准号: 2713771
• 金额: --
• 依托单位: Durham University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2713771
• 关键词: Neuronal; basis; pheromone; perception; mosquitoes

Motivation for the project. Anopheles mosquitoes are the vectors of malaria that affects half of the world population (WHO 2019). Mosquitoes heavily rely on their sense of smell to find a human for a blood meal (Konopka et al, 2021). Surprisingly, reports of chemical communication in Anopheles are very scarce and recent, and the use of pheromones is not universally established for any mosquitoes. In contrast to mosquitoes, most other insects are well known to use volatile and non-volatile pheromones (Blomquist and Bagnerés, 2010). We were intrigued by the obvious lack of information about mosquito pheromones, and thus the implicit assumption that mosquitoes, in contrast to other insects, don't produce and use chemical cues to communicate intraspecifically. We employed novel methods and techniques for our pilot studies, and identified 17 potential male and female pheromone candidates in Anopheles gambiae. This project will verify and strengthen our preliminary findings, and will explore how Anopheles gambiae respond to pheromone candidates at the levels of cells, organs and the entire organism. We will employ cutting edge techniques of insect neuroscience and genetic engineering to achieve our aims. Aims. 1) To verify pheromone candidates and identify olfactory receptors that detect pheromone candidates in male and female mosquitoes; 2) To create a transgenic mosquito line targeting neurons that specifically respond to the pheromones; 3) To genetically manipulate the pheromone neurons and test their effect on mosquito mating behaviours. Methodology. Pheromone candidates will be verified by GC/MS coupled with thermodesorption. Olfactory receptors will be identified via live calcium imaging from mosquito and Drosophila olfactory neurons. New transgenic lines will be generated by using CRISPR/Cas9 to replace the stop codon of the candidate OR genes with a T2A-QF2 construct. Panel of olfactory preference, flight activity and mating and behavioural assays will be developed and conducted on male and female An.gambiae. Timetable. Year1: Aim 1 in Durham and Wuerzburg; conference ECRO/ESITO in Europe; Year2: Aims 1 and 2 in Durham; SfN conference/Neuroethology Congress; Optical Imaging and Electrophysiological Recordings in Neuroscience School, Paris; Year3: Aim 3 in Newcastle and Durham; PIPS placement; ESA conference; Year4: MBL course "Neural System and Behavior", Woods Hole, USA; finishing up experiments and writing up. Novelty. This project will involve cutting edge techniques of insect neuroethology, neuroscience and genetics that the supervisors are world leaders in. In addition, understanding mosquito sexual communication has profound implications for mosquito and malaria control.

项目的动机。按蚊是影响世界一半人口的疟疾病媒（WHO 2019）。蚊子在很大程度上依赖于它们的嗅觉来寻找人类的血餐（Konopka等人，2021）。令人惊讶的是，有关按蚊化学通讯的报道非常罕见，而且是最近才有的，而且信息素的使用并没有普遍建立在任何蚊子身上。与蚊子相反，众所周知，大多数其他昆虫使用挥发性和非挥发性信息素（Blomquist和Bagnerés，2010）。我们对明显缺乏关于蚊子信息素的信息很感兴趣，因此隐含的假设是，与其他昆虫相比，蚊子不会产生和使用化学线索来进行种内交流。我们采用新的方法和技术，我们的试点研究，并确定了17个潜在的男性和女性信息素候选人冈比亚按蚊。该项目将验证和加强我们的初步研究结果，并将探讨冈比亚按蚊如何在细胞，器官和整个生物体的水平上对信息素候选物作出反应。我们将采用昆虫神经科学和基因工程的尖端技术来实现我们的目标。 目标。1)验证信息素候选物并鉴定检测雄性和雌性蚊子中的信息素候选物的嗅觉受体; 2）创建靶向对信息素特异性响应的神经元的转基因蚊子系; 3）遗传操纵信息素神经元并测试它们对蚊子交配行为的影响。 方法论信息素候选物将通过GC/MS结合热解吸进行验证。嗅觉受体将通过蚊子和果蝇嗅觉神经元的活体钙成像来鉴定。将通过使用CRISPR/Cas9用T2 A-QF 2构建体替换候选OR基因的终止密码子来产生新的转基因系。将对雄性和雌性冈比亚按蚊进行嗅觉偏好、飞行活动、交配和行为测定。 时间表第一年：目标1在达勒姆和维尔茨堡;欧洲ECRO/ESITO会议;第2年：目标1和2在达勒姆; SfN会议/神经行为学大会;巴黎神经科学学院的光学成像和电生理记录;第3年：目标3在纽卡斯尔和达勒姆; PIPS安置; ESA会议;第4年：MBL课程“神经系统和行为”，伍兹霍尔，美国;完成实验和写作。 新奇。该项目将涉及昆虫神经行为学，神经科学和遗传学的尖端技术，这些技术的主管是世界领先的。此外，了解蚊子的性交流对蚊子和疟疾的控制具有深远的意义。