Collaborative Research: The genetic basis of call diversity in acoustic insects

动物的行为在生物多样性中起着重要的作用。例如，许多昆虫有特定的叫声，这是物种之间交配的障碍。这种多样性在很大程度上是由基因控制的。发育过程中的基因表达和调控模式也会导致特定的行为。然而，目前还不清楚行为差异与遗传差异的关系：相对于基因调控的变化，实际基因的变化有什么贡献？这仍然是一个具有普遍重要性的基本开放问题，远远超出了昆虫的不同叫声。在这项研究中，这个问题是解决在一个Katydid物种，产生两个替代的呼吁，在发展过程中所经历的环境。在一个种群中，这些叫声之间的差异是由于基因调控的差异。种群水平的差异也发生在这种昆虫的地理范围内，这种变化主要是由个体基因的差异引起的。这使得基因和遗传调控的贡献，在这个物种的行为多样性的评估。该项目整合了遗传学，神经生物学，行为学和进化。其成果有助于了解生物多样性的遗传基础，并与动物行为以外的许多领域有关，包括医学研究。更广泛的影响包括为不同背景的学生提供培训机会，使他们为生命科学职业的挑战做好准备。外展活动在一个相关的研究系统中向公众介绍现代生物研究。该项目的目标是了解Neoconocephalus katydids的呼叫多样性的遗传基础。物种Neoconocephalus triops经历了发育可塑性（DP），导致两个替代的广告呼叫与两个时间特性（脉冲率和诗句模式）的物种水平差异。DP可以通过改变性成熟前的光周期来可靠地诱导。首先，将确定CNS中有助于产生呼叫模式的相关发育阶段和组织（中枢模式发生器，CPG）。目的1使用损伤实验、钙成像和选择性加热单个神经节来确定CPG的位置。在目标2中，研究人员通过改变短日照条件下的暴露时间和分析成虫的叫声表型来确定表型诱导的敏感期。目标3探索了呼叫模式多样性的遗传基础。将使用在目的2中确定的相关时间采集的含有CPG的组织进行RNAseq。 将确定表型之间的相关性和基因表达模式的差异以及种群间的遗传差异。比较包括两个呼叫表型的三个人口，显着不同的呼叫模式和程度的DP。在目标4中，研究人员将使用RNAi操纵目标3中确定的候选基因的表达水平，并检查呼叫表型的后果，从识别相关性转向测试因果关系。结果提供了可检验的假说的遗传控制的呼叫变化，这起着重要的作用，这组的多样化。 培训和外展活动的重点是MO，KY和WV农村地区的公众和学生，这是传统上服务不足的群体。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The behavior of animals plays an important role in biological diversity. For example, many insects have specific calls which are barriers to mating between species. This diversity is in large part under the control of genes. Patterns of gene expression and regulation during development can also result in specific behaviors. However, it is unclear how differences in behavior relate to genetic differences: what is the contribution of changes in the actual genes relative to that of changes in gene regulation? This remains a fundamental open question of general importance, far beyond the diverse calls of insects. In this research this question is addressed in a katydid species that produces two alternative calls in response to the environment experienced during development. The differences between these calls within a population are due to differences in gene regulation. Population-level differences also occur across the geographic range of this insect, and this variation is largely caused by differences in individual's genes. This allows the assessment of the contributions of both genes and genetic regulation to the diversity of behavior in this species. This project integrates genetics, neurobiology, behavior, and evolution. Its outcomes contribute to the understanding of the genetic foundation of biodiversity and are relevant in many areas outside of animal behavior, including medical research. Broader impacts include training opportunities for students of diverse backgrounds that prepare them for the challenges of careers in life sciences. Outreach activities introduce the public to modern biology research in a relatable study system. The goal of this project is to understand the genetic underpinnings of the call diversity in Neoconocephalus katydids. The species Neoconocephalus triops undergoes developmental plasticity (DP) that results in two alternative advertisement calls with species-level differences in two temporal properties (pulse rate and verse pattern). DP can be induced reliably by varying the photoperiod before sexual maturation. First, the relevant developmental stages and tissues (central pattern generators, CPG) in the CNS that contribute to the call pattern generation will be identified. Aim 1 identifies the location of the CPGs using lesion experiments, calcium imaging, and selectively heating of individual ganglia. In aim 2, the investigators identify the sensitive phase of phenotype induction by varying the exposure time of short day conditions and analyzing adult call phenotypes. Aim 3 explores the genetic underpinnings of the diversity of call patterns. RNAseq will be conducted using tissues containing the CPGs collected at the relevant times identified in aim 2. Correlations between phenotypes and differences in gene expression patterns as well as genetic differences among populations will be identified. Comparisons include two call phenotypes of three populations that differ significantly in call pattern and extent of DP. In aim 4, the investigators will manipulate the expression levels of candidate genes identified in aim 3, using RNAi and examine the consequences for call phenotypes, moving from identifying correlations to testing causation. The outcomes provide testable hypotheses about the genetic control of call variation, which plays a significant role in the diversification of this group. Training and outreach activities are focusing on the public and students of rural areas in MO, KY, and WV, which are traditionally underserved groups.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.