Cell lineage-based investigation of chemosensory neuron development

基于细胞谱系的化学感应神经元发育研究

基本信息

批准号：
10373822
负责人：
Pavak Kirit Shah
金额：
$ 23.86万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10373822
关键词：
3-Dimensional Ablation Anatomy Animals Behavior Behavior Control Behavioral Biological Assay Caenorhabditis elegans Cell Lineage Cell Nucleus Cell membrane Cells Chemotaxis Computer software Cues Development Developmental Biology Embryo Embryonic Development Evolution Fluorescence Microscopy Future Gene Expression Gene Transfer Techniques Genetic Genetic Screening Genetic Transcription Health Hookworms Human Image Individual Invertebrates Investigation Label Lasers Lead Light Link Maps Modeling Morphology Nematoda Neuroanatomy Neurons Nuclear Odors Order Coleoptera Organism Parasites Parasitic nematode Pathway interactions Pattern Persons Pheromone Physiology Prophylactic treatment Regulation Resolution Resource-limited setting Sensory Signal Transduction Skin Specific qualifier value Strongyloides stercoralis Systems Development Taste Perception Time Training Work base blastomere structure body position cellular imaging contrast imaging convolutional neural network design developmental genetics fluorescence imaging genetic manipulation insight light microscopy microscopic imaging neglected tropical diseases neural circuit neural network neuron development novel programs reconstruction relating to nervous system response social tool treatment strategy vertebrate embryos

项目摘要

Project Summary Signaling from chemosensory neurons regulates changes in animal physiology and behavior in response to environmental and social cues. Sensory neuroanatomy is so broadly conserved in nematodes that, based on morphology and cell body position, functionally homologous chemosensory neurons have been identified across widely divergent nematode genera, including the well-studied free living nematode Caenorhabditis elegans, the skin-penetrating human parasite Strongyloides stercoralis, and the predatory nematode Pristionchus pacificus. Despite this homology, little is known about the conservation of the developmental and genetic programs that produce individual chemosensory neurons and maintain or differentiate their function. To what extent do anatomically homologous neurons share conserved chemosensory function? And to what extent does anatomical homology reflect a common developmental program? We will answer these questions by mapping the cell lineages that give rise to chemosensory neurons, determining the extent to which positionally homologous chemosensory neurons are specified by conserved transcriptional regulators, and identifying conserved chemosensory function. We will achieve this by developing a novel 3D style transfer convolutional neural network (stCNN) to automate the identification of major cellular features such as the nucleus and cell membrane in transmitted light imaging with differential interference contrast (DIC). We will then use this tool to reconstruct the embryonic lineages of S. stercoralis and P. pacificus, map the expression of known regulators of chemosensory neural identity to these lineages, and assess the conservation of function between homologous chemosensory neurons by performing laser cell ablations and single-worm chemotaxis assays. This work has direct relevance to human health, since chemosensation regulates many aspects of development, physiology, and behavior in S. stercoralis and other human-parasitic nematodes. Parasitic nematodes infect over a billion people worldwide and cause some of the most common and devastating neglected tropical diseases, particularly in low-resource settings. Our multi-species approach will allow us to determine which aspects of nematode chemosensory system development and function are broadly conserved, and which contain species- specific adaptations that drive species-specific behaviors, including parasitic behaviors. Furthermore, the automated reconstruction of cell lineages from DIC images will be an enabling tool of broad value. The ability to map new developmental lineages without transgenesis will be especially transformative in the study of human-parasitic nematodes such as hookworms that are not amenable to genetic manipulation, and can be extended to non-nematode species, including early-stage vertebrate embryos.

项目概要来自化学感应神经元的信号调节动物生理和行为的变化以响应环境和社会线索。感觉神经解剖学在线虫中如此广泛保守，基于已鉴定出形态和细胞体位置、功能同源的化学感觉神经元跨越广泛不同的线虫属，包括经过充分研究的自由生活线虫 Caenorhabditis 线虫、穿透皮肤的人体寄生虫粪类圆线虫和掠食性线虫 Pristionchus 太平洋。尽管存在这种同源性，但人们对发育和遗传程序的保护知之甚少。产生单个化学感应神经元并维持或分化其功能。做到什么程度解剖上同源的神经元共享保守的化学感应功能？以及到什么程度解剖同源性反映了共同的发育程序？我们将通过映射来回答这些问题产生化学感应神经元的细胞谱系，决定了位置上的程度同源化学感应神经元由保守的转录调节因子指定，并识别保守的化学感应功能。我们将通过开发一种新颖的 3D 风格传输卷积来实现这一目标神经网络 (stCNN) 自动识别主要细胞特征，例如细胞核和细胞具有微分干涉衬度 (DIC) 的透射光成像中的膜。然后我们将使用这个工具重建粪圆线虫和太平洋圆线虫的胚胎谱系，绘制已知调节因子的表达图这些谱系的化学感应神经同一性，并评估同源之间的功能保守性通过进行激光细胞消融和单蠕虫趋化测定来观察化学感应神经元。这项工作与人类健康直接相关，因为化学感觉调节发育的许多方面，粪圆线虫和其他人类寄生线虫的生理学和行为。寄生线虫感染超过全世界有十亿人，导致了一些最常见和最具破坏性的被忽视的热带疾病，特别是在资源匮乏的环境中。我们的多物种方法将使我们能够确定哪些方面线虫化学感应系统的发育和功能广泛保守，并且包含物种- 驱动物种特定行为的特定适应，包括寄生行为。此外，根据 DIC 图像自动重建细胞谱系将成为具有广泛价值的工具。能力在没有转基因的情况下绘制新的发育谱系将在研究中特别具有变革性人类寄生线虫，例如钩虫，不适合基因操作，可以通过扩展到非线虫物种，包括早期脊椎动物胚胎。