Gene regulatory mechanisms required for neuronal chemosensitivity

神经元化学敏感性所需的基因调控机制

• 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
• 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

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.

项目总结/摘要： 哺乳动物脑干的化学敏感区调节呼吸，并受到呼吸的小幅度增加的刺激。 动脉血中的二氧化碳（CO2）水平。这种化学敏感性的遗传和分子基础很差 明白C.线虫是探索这个问题的一个很好的遗传模型，因为它具有对CO2敏感的 神经元-BAG神经元-介导刻板行为，其细胞生理学可以在体内研究。 BAG细胞命运由ETS-5（一种ETS结构域转录因子）决定。引人注目的是，哺乳动物同源物 ETS-5的Pet 1是CO2化学敏感脑区发育所必需的，Pet 1突变体具有 调节呼吸以应对二氧化碳挑战的缺陷。寻找在C. elegans和可能在脊椎动物中，我使用ChIP-seq分析了ETS-5的直接转录靶点。那么 为了阐明这些靶点中哪些在BAG中直接受ETS-5调节，我对野生型进行了mRNA-seq 与ets-5突变的BAG神经元相比。为了确定这些目标中的哪些对CO2具有重要的功能， 我对二氧化碳回避缺陷进行了行为筛查。RGS-6是一种G蛋白激活蛋白， 直接ETS-5转录靶，其似乎在ETS-5突变体背景下下调，和rgs-6 突变也会导致回避缺陷。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置于GPCR中， （3）确定RGS-6在CO2传感中的生理作用。拟议的研究将结合 分子遗传学，基因组学和体内功能成像，以阐明一种新的RGS蛋白在CO2中的作用 感应由于Pet 1样因子在C.我们的研究可能会阐明 脑中化学敏感神经元的功能所需的机制，其在调节 呼吸功能障碍与致命的呼吸暂停有关。