Development of optogenetically controlled gene expression tools for the characterization of neuronal circuits involved in insect reproduction

开发光遗传学控制的基因表达工具，用于表征昆虫繁殖中涉及的神经元回路

基本信息

批准号：
BB/N021827/1
负责人：
Matthias Soller
金额：
$ 19.2万
依托单位：
University of Birmingham
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2017
资助国家：
英国
起止时间：
2017 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FN021827%2F1
关键词：
Development optogenetically controlled gene expression

项目摘要

Reproductive behaviors and their regulation are most fundamental to all animals, but have been exploited little for population control in insects. Since they are hard-wired into the brain we can learn how this behavioral program is encoded in the brain and shaped by perception and decision-making processes. Understanding how behavior is encoded in the brain is one of the big challenges in biology and requires a behaviorally and genetically tractable model organism, but also tools to manipulate localized neurons. One of the key tools to achieve this aim are light-manipulateable molecules, such ion channels, where light can be used to control neuronal activity in space and time. Due to the small size of insects, however, this technology has its limitation. Here, we want to adapt light-inducible transcription factors derived from bacteria and plants already used in mammalian cell culture to Drosophila to characterize the neuronal circuits involved in reproduction. For this analysis we will capitalize on gene expression regulatory sequences known to characterize neuronal populations involved in reproduction, but these gene expression patterns are complex. To be able to assign functions to localized neurons therefore requires spatial dissection of these gene expression patterns, which can be achieved by light-controlled transcription factors.To develop such light-controlable tools to manipulate gene expression, we will capitalize on the robust post-mating responses (PMRs) of the fruit fly Drosophila melanogaster. Here, male-derived sex-peptide (SP) transferred during mating is the key molecule leading to refusal to remate and induction of egg laying. The very robust behavioral response of Drosophila females to sex-peptide provides the essential prerequisites to map SP responsive neurons and eventually learn how complex behaviors such as mating choice and control of egg laying are encoded in the brain.Our recent studies showed that there are several distinct neuronal populations that can via exposure to SP induce refusal to remate and egg laying. We currently do not know where in the fly these neurons are located, however, we could show that these two post-mating responses can be separated. Candidate neurons for SP induced post-mating responses include sensory neurons in the genital tract and in the legs, but also neurons in the abdominal ganglion and the central brain. To identify the neuronal circuitry underlying the sex-peptide response, we will use light induced gene expression directed to neurons in specific parts of the female fly body to express membrane-tethered SP. Such optogentic manipulation of gene expression has the advantage to be fully controllable in space and time. With these experiments we will test the hypothesis that the response to SP is comprised of a modular assembly of individual elements, e.g. refusal to remate or induction of egg laying. Compared to the previous model arguing for central induction of all PMRs, a modular assembly of individual PMRs holds evolutionary flexibility during speciation and adaptation to diverse habitats, but can maintain basic regulatory principles such as the control of egg laying. We therefore anticipate that the knowledge obtained from our studies will be applicable to a wide range of pest insects pinpointing towards novel strategies for pest management to protect crop and control insect born diseases by interfering with egg laying. In particular, our findings are directly transferable to the close relative Drosophila suzukii, one of the few species able to lay eggs into fruits, which is currently invading Europe including the UK and causing damage worth billions of pounds to fruit production.

生殖行为及其调节对所有动物都是最基本的，但对昆虫的种群控制很少被利用。由于它们是在大脑中刻连接的，因此我们可以了解该行为程序如何在大脑中编码并受感知和决策过程的影响。了解在大脑中如何编码行为是生物学中最大的挑战之一，需要在行为和遗传上可触犯的模型生物，也需要操纵局部神经元的工具。实现此目标的关键工具之一是可轻应化的分子，例如离子通道，可以使用光来控制时空中的神经元活动。但是，由于昆虫的尺寸很小，因此该技术具有其限制。在这里，我们要适应源自哺乳动物细胞培养的细菌和植物衍生出的光诱导转录因子，以表征涉及生殖的神经元回路。在此分析中，我们将利用已知的基因表达调节序列，以表征与生殖有关的神经元种群，但这些基因表达模式很复杂。因此，要能够将功能分配给局部神经元，需要对这些基因表达模式进行空间解剖，这可以通过光控制的转录因子来实现。要开发这种可控制的工具来操纵基因表达，我们将资本利用鲁棒的post术后反应（PMR）的果蝇蝇蝇Drosophila Melanogaster。在这里，在交配过程中转移的男性性肽（SP）是导致拒绝和诱导产卵的关键分子。果蝇女性对性肽的行为反应非常强大，为绘制SP响应性神经元的映射提供了必要的先决条件，并最终了解了在大脑中编码诸如配合选择和卵子铺设的复杂行为，例如在大脑中编码。我们目前不知道这些神经元在哪里，但是，我们可以证明可以分开这两种论到这两个后的响应。 SP诱导后反应的候选神经元包括生殖道和腿部的感觉神经元，以及腹部神经节和中央大脑中的神经元。为了识别性肽反应的基础的神经元电路，我们将使用针对雌性蝇体特定部位的神经元的光诱导基因表达来表达膜螺旋状的SP。这种对基因表达的选择性操纵具有在空间和时间上完全可控制的优势。通过这些实验，我们将检验以下假设：SP的响应由单个元素的模块化组成，例如拒绝解除或诱导产卵。与先前争论所有PMR的中心诱导的模型相比，单个PMR的模块化组件在物种形成和适应各种栖息地的过程中具有进化柔韧性，但可以维持基本的调节原理，例如控制卵的控制。因此，我们预计从我们的研究中获得的知识将适用于广泛的害虫昆虫，这些昆虫指向有害生物管理的新型策略，以通过干扰产卵来保护作物和控制昆虫的疾病。特别是，我们的发现可直接转移到近亲相对果蝇铃木，这是少数能够将鸡蛋产卵的物种之一，目前正在入侵包括英国在内的欧洲，并造成价值数十亿英镑的水果生产的损失。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Channel nuclear pore complex subunits are required for transposon silencing in Drosophila.

DOI：
10.7554/elife.66321
发表时间：
2021-04-15
期刊：
eLife
影响因子：
7.7
作者：
Munafò M;Lawless VR;Passera A;MacMillan S;Bornelöv S;Haussmann IU;Soller M;Hannon GJ;Czech B
通讯作者：
Czech B

Channel nuclear pore protein 54 directs sexual differentiation and neuronal wiring of female reproductive behaviors in Drosophila.

DOI：
10.1186/s12915-021-01154-6
发表时间：
2021-10-20
期刊：
BMC biology
影响因子：
5.4
作者：
Nallasivan MP;Haussmann IU;Civetta A;Soller M
通讯作者：
Soller M

Indel driven rapid evolution of core nuclear pore protein gene promoters.

DOI：
10.1038/s41598-023-34985-0
发表时间：
2023-05-17
期刊：
Scientific reports
影响因子：
4.6
作者：
通讯作者：

Thiamethoxam exposure deregulates short ORF gene expression in the honey bee and compromises immune response to bacteria.

DOI：
10.1038/s41598-020-80620-7
发表时间：
2021-01-15
期刊：
Scientific reports
影响因子：
4.6
作者：
Decio P;Ustaoglu P;Derecka K;Hardy ICW;Roat TC;Malaspina O;Mongan N;Stöger R;Soller M
通讯作者：
Soller M