Genetically engineered ants to label and study neurons involved in social behavior

基因工程蚂蚁可以标记和研究参与社会行为的神经元

• 批准号: 10370381
• 负责人: Roberto Bonasio
• 金额: $ 20.31万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370381
• 关键词: Adult; Affect; Alleles; Animals; Ants; Behavior; Behavioral; Biological Models; Brain; Brain imaging; CRISPR/Cas technology; Castes; Cell Count; Cells; Chemicals; Cognition Disorders; Data; Defect; Development; Dissection; Engineering; Environment; Epigenetic Process; Event; Future; Gene Expression Regulation; Gene Transfer Techniques; Generations; Genetic; Genetic Engineering; Genetic Recombination; Genome; Goals; Gonadotropin Hormone Releasing Hormone; Head; Homologous Gene; Human; Individual; Injections; Insecta; Knock-in; Label; Lead; Mammals; Mediator of activation protein; Membrane; Molecular; Morphology; Nerve Degeneration; Neuroglia; Neurons; Neuropeptides; Pathway interactions; Phenotype; Physiology; Play; Process; Property; Regulation; Reporter; Reporting; Reproduction; Role; Signal Transduction; Social Behavior; Social status; Specific qualifier value; Stimulus; Structure; Synapses; System; Transgenic Organisms; Vasopressin Receptor; Vasopressins; Work; behavioral plasticity; brain remodeling; differential expression; epigenetic regulation; experimental study; flexibility; genetic approach; genetic manipulation; mind control; neuronal circuitry; next generation; promoter; receptor; reproductive; response; single-cell RNA sequencing; social; tool

ABSTRACT The goal of this proposal is to generate genetic reporter lines for neuronal circuits that regulate social behavior in Harpegnathos saltator ants and to utilize these lines to visualize brain remodeling as adult workers reprogram themselves to behave as ant queens. Social insects are an emerging model system to study the epigenetic regulation of brain function and behavior because the same genome specifies alternative behavioral states in different castes. Whether and how brains of workers and queens are differently wired to implement distinct sets of caste-specific behaviors is not known. In Harpegnathos ants, adult workers can become queens via a caste transition that involves dramatic phenotypic changes in reproduction, physiology, and behavior. This feature of Harpegnathos ants allows for easy propagation of engineered alleles, as any individual can become reproductive. The ability of adult Harpegnathos workers to become queens offers unique opportunities for the mechanistic dissection of adult brain plasticity. Single-cell RNA-seq analyses revealed major changes in cellular composition affecting both neurons and glia, indicating that a structural remodeling of the brain accompanies the caste transition. The molecular signals that direct this brain remodeling remain unknown. We previously showed that the neuropeptide corazonin stimulates hunting, a worker-specific behavior, and is downregulated as workers become queens. We have since obtained new preliminary evidence that the ant homolog of human vasopressin is expressed in a caste-specific manner and likely also regulates a subset of social behaviors in Harpegnathos ants. We hypothesize that signaling by these neuropeptides directs brain remodeling events that underpin the switch in social behaviors during the caste transitions. In Aim 1, we will utilize a transgenic approach to label corazonin- and vasopressin-producing neurons and their projection with membrane-bound GFP and to observe their remodeling during the natural caste transition. In Aim 2, we will utilize CRISPR/Cas9 to generate knock-in driver and reporter lines to label neurons that respond to corazonin or vasopressin and visualize their plasticity at the cellular and synaptic level in response to the neuropeptides. These experiments will reveal crucial information on adult brain plasticity in Harpegnathos ants and provide sophisticated genetic tools for the further dissection of the epigenetic regulation of social behavior in these ants. Given that corazonin and vasopressin have mammalian counterparts, our results are expected to have a broad impact on our understanding of how neuropeptides regulate brain plasticity and social behavior.

抽象的 该提案的目标是为调节社会的神经元回路生成遗传报告系 Harpegnathos salator ant 的行为，并利用这些线条来可视化成年工蚁的大脑重塑 重新编程自己，使其表现得像蚁后。 社会性昆虫是研究大脑功能和行为的表观遗传调控的新兴模型系统 因为相同的基因组指定了不同种姓的不同行为状态。是否以及如何大脑 工蚁和蚁后执行不同种姓特定行为的方式不同，目前尚不清楚。 在 Harpegnathos 蚂蚁中，成年工蚁可以通过种姓转变成为蚁后，这种转变涉及戏剧性的过程 生殖、生理和行为的表型变化。 Harpegnathos 蚂蚁的这一特征使得 工程等位基因易于繁殖，因为任何个体都可以繁殖。 成年 Harpegnathos 工人成为女王的能力为机械师提供了独特的机会 成人大脑可塑性的解剖。单细胞 RNA-seq 分析揭示了细胞的重大变化 影响神经元和神经胶质细胞的成分，表明大脑的结构重塑伴随着 种姓过渡。指导这种大脑重塑的分子信号仍然未知。 我们之前表明，神经肽 corazonin 会刺激狩猎，这是一种工人特有的行为，并且 当工人成为女王时，其监管就会下调。此后我们获得了新的初步证据表明蚂蚁 人类加压素的同源物以种姓特异性方式表达，并且可能还调节一个子集 Harpegnathos 蚂蚁的社会行为。我们假设这些神经肽的信号传导引导大脑 在种姓转变期间支撑社会行为转变的重塑事件。 在目标 1 中，我们将利用转基因方法来标记产生 corazonin 和 vasopressin 的神经元，并 他们用膜结合的 GFP 进行投影，并观察他们在自然种姓转变过程中的重塑。 在目标 2 中，我们将利用 CRISPR/Cas9 生成敲入驱动程序和报告基因线来标记神经元 对 corazonin 或加压素做出反应，并在细胞和突触水平上可视化它们的可塑性 到神经肽。 这些实验将揭示关于 Harpegnathos 蚂蚁成年大脑可塑性的重要信息，并提供 复杂的遗传工具，用于进一步剖析这些群体中社会行为的表观遗传调控 蚂蚁。鉴于考拉佐宁和加压素在哺乳动物中具有对应物，我们的结果预计将具有 对我们对神经肽如何调节大脑可塑性和社会行为的理解产生广泛影响。