Gene regulatory mechanisms required for neuronal chemosensitivity

神经元化学敏感性所需的基因调控机制

基本信息

批准号：
10379754
负责人：
Mary Grace Rossillo
金额：
$ 0.25万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
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：
2021-03-30 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10379754
关键词：
Acute Affect Animal Model Animals Apnea Area Auxins Behavior Behavioral Binding Biochemical Blood Brain Brain Stem Brain region Breathing Caenorhabditis elegans Calcium Carbon Dioxide Cell physiology Cells ChIP-seq Cyclic GMP Defect Development ETS Family Protein Environment Family Fluorescence Functional Imaging Functional disorder G-Protein-Coupled Receptors GTP-Binding Protein Regulators GTP-Binding Protein alpha Subunits, Gs GTP-Binding Proteins GTPase-Activating Proteins Generations Genetic Genetic Models Genetic Transcription Genomics Guanosine Triphosphate Heterotrimeric GTP-Binding Proteins Homologous Gene Human Hydrolysis Hypercapnia Invertebrates Link Location Mammals Measures Mediating Methods Modality Molecular Molecular Genetics Monitor Motor Mutate Mutation Nematoda Neonatal Mortality Neurons Pathway interactions Pattern Physiological Physiological Processes Physiology Play Proteins RGS Family Gene RGS Proteins Regulator Genes Role Second Messenger Systems Signal Transduction Signaling Protein Site Stereotyped Behavior Stimulus Structure Sudden infant death syndrome System Time Vertebrates base behavioral response chromatin immunoprecipitation experimental study genome editing in vivo mRNA sequencing member mutant nematode genetics nervous system disorder neurochemistry neurotransmitter release next generation sequencing novel programs raphe nuclei receptor receptor function relating to nervous system respiratory respiratory gas response transcription factor

项目摘要

PROJECT SUMMARY / ABSTRACT: Chemosensitive areas of the mammalian brainstem regulate breathing and are stimulated by small increases in carbon dioxide (CO2) levels in arterial blood. The genetic and molecular bases for this chemosensitivity are poorly understood. C. elegans is an excellent genetic model for probing this question because it possesses CO2-sensitive neurons - the BAG neurons - that mediate stereotyped behavior and whose cell physiology can be studied in vivo. BAG cell fate is determined by ETS-5, an ETS-domain transcription factor. Strikingly, the mammalian homolog of ETS-5, Pet1, is required for the development of CO2-chemosensitive brain regions, and Pet1 mutants have defects regulating breathing in response to CO2 challenges. To find molecules important in CO2 sensing in C. elegans and perhaps in vertebrates, I analyzed the direct transcriptional targets of ETS-5 using ChIP-seq. Then, to clarify which of these targets are being directly regulated by ETS-5 in BAG, I performed mRNA-seq on wild type versus ets-5 mutant BAG neurons. To determine which of these targets are functionally important for CO2 sensing, I performed a behavioral screen for CO2 avoidance defects. RGS-6 is a G-protein activating protein that is a direct ETS-5 transcriptional target that appears to be down-regulated in an ets-5 mutant background, and rgs-6 mutation also results in an avoidance defect. To better understand the role of RGS-6 in CO2 sensing, I will (1) determine the expression pattern, site, and time of action of rgs-6 in CO2 sensing, (2) place RGS-6 in a GPCR pathway, and (3) determine the physiological role of RGS-6 in CO2 sensing. The proposed studies will integrate molecular genetics, genomics, and in vivo functional imaging to elucidate the role of a novel RGS protein in CO2 sensing. Because Pet1-like factors are conserved between C. elegans and humans, our studies will likely elucidate mechanisms required for the function of chemosensitive neurons in the brain, which play critical roles in regulating breathing and whose dysfunction is linked to fatal apneas.

项目摘要 /摘要：哺乳动物脑干的化学敏感区域调节呼吸，并受到较小的增加的刺激动脉血的二氧化碳（CO2）水平。这种化学敏感性的遗传和分子碱基较差理解。秀丽隐杆线虫是探测这个问题的绝佳遗传模型，因为它具有二氧化碳敏感的神经元 - 袋神经元 - 介导刻板印象的行为，可以在体内研究其细胞生理学。袋细胞命运由ETS-5（ETS域转录因子）确定。令人惊讶的是，哺乳动物同源物 co2化学敏感大脑区域的发展需要ETS-5，PET1，PET1突变体具有响应二氧化碳挑战以调节呼吸的缺陷。在C中找到对CO2感应重要的分子。秀丽隐杆线，也许是在脊椎动物中，我使用CHIP-SEQ分析了ETS-5的直接转录靶标。然后，澄清这些目标中的哪一个是由袋子中的ETS-5直接调节的，我对野生型进行了mRNA-Seq 对ETS-5突变袋神经元。确定这些目标中的哪个对CO2在功能上很重要感知，我对二氧化碳避免缺陷进行了行为屏幕。 RGS-6是一种G蛋白激活蛋白，是一种直接ETS-5转录靶标在ETS-5突变体背景中似乎下调，RGS-6 突变还会导致避免缺陷。为了更好地了解RGS-6在CO2传感中的作用，我将（1）确定RGS-6在CO2传感中的表达模式，位点和作用时间，（2）将RGS-6放在GPCR中途径，（3）确定RGS-6在CO2传感中的生理作用。拟议的研究将整合分子遗传学，基因组学和体内功能成像，以阐明新型RGS蛋白在CO2中的作用感应。因为秀丽隐杆线虫和人类之间的pet1样因素是保守的，所以我们的研究可能会阐明大脑中化学敏感性神经元功能所需的机制，该神经元在调节中起关键作用呼吸，其功能障碍与致命的呼吸暂停有关。