Metabolic coupling of neuronal ion transport

神经元离子转运的代谢耦合

• 批准号: 10155101
• 负责人: Dylan John Meyer
• 金额: $ 6.6万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10155101
• 关键词: Active Biological Transport; Active Ion Transport; Acute; Address; Affect; Bioinformatics; Biosensor; Brain; Buffers; CKB gene; CRISPR/Cas technology; Carbohydrates; Carrier Proteins; Cell membrane; Cells; Consumption; Coupled; Couples; Coupling; Creatine Kinase; Cytoplasmic Granules; Data; Data Analyses; Dependence; Development; Dyes; Electrophysiology (science); Environment; Enzymes; Epilepsy; Event; Feedback; Fellowship; Functional disorder; Genes; Glycolysis; High Fat Diet; Hippocampus (Brain); Homeostasis; Immersion; Individual; Ion Channel; Ion Transport; Ions; Isoenzymes; Journals; Knock-out; Knockout Mice; Knowledge; Leadership; Learning; Measures; Mediating; Membrane; Metabolic; Metabolic Pathway; Microscopy; Mitochondria; Na(+)-K(+)-Exchanging ATPase; Neurodegenerative Disorders; Neurons; Oxidative Phosphorylation; Pathology; Pharmacology; Process; Production; Pump; RNA; Reaction; Reducing diet; Research; Rest; Role; Signal Transduction; Slice; Source; Synapses; Therapeutic; Time; Tissues; Training; Transport Process; Work; Writing; adenylate kinase; brain cell; career; experience; experimental study; fluorescence lifetime imaging; inhibitor/antagonist; innovation; ion dynamics; ketogenic diet; medical schools; mitochondrial metabolism; neural circuit; neuronal excitability; neuronal metabolism; neurotransmission; postsynaptic; receptor; response; sensor; skills; two-photon; voltage gated channel

Proper energy utilization and management by the brain is essential for neurons to process information and communicate effectively. It is well known that active ion transport activities consume enormous quantities of ATP during neuronal signaling to restore plasma membrane ion gradients and maintain cellular excitability, but how active ion transport is fueled by specific metabolic pathways and ATP buffering mechanisms is still controversial. This research fellowship aims to study two aspects of energy management during neuronal signaling: 1) whether active ion transport preferentially couples to ATP produced from glycolysis or from oxidative phosphorylation, and 2) whether creatine kinase and adenylate kinase buffer ATP during periods of intense energy demand. The proposed experiments will utilize two-photon fluorescence lifetime imaging of genetically encoded fluorescent biosensors and dyes to accurately quantify real-time metabolite and ion dynamics in hippocampal dentate granule neurons of acute brain slices following synaptic stimulation. Pharmacological strategies will then be employed to tease apart the contributions of specific active transport activities, ion channels, and metabolic pathways to the metabolite and ion sensor lifetime signals. Also, CRISPR-Cas9:sg RNA gene editing will be used to determine how ATP buffering in dentate granule neurons is affected by knockout of creatine kinase or adenylate kinase isozymes. Knowing whether neuronal active ion transport during signaling is differentially regulated by glycolysis or oxidative phosphorylation, and whether it is supported by ATP-buffering enzymes, is important for detailing how neuronal excitability is regulated by different metabolic fuels, and will have implications for understanding how the metabolic alterations resulting from a ketogenic diet – a very-low- carbohydrate, high-fat diet – are therapeutic for epilepsy. This knowledge will also help to determine the underlying pathophysiological mechanisms of neurodegenerative disorders that are associated with energetic dysfunction. This fellowship training plan contains numerous outside-the-lab activities to aid the awardee's scientific development and allow for continued learning, including courses on neural circuits, microscopy, bioinformatics skills for data analysis, scientific writing, lab leadership, and many others. Weekly department seminars and journal clubs will allow the awardee to learn about related research and receive feedback about their data and hypotheses. The research environment of Harvard Medical School will provide the awardee with an immersive learning and training experience that will facilitate their transition into the next stage of their career.

大脑适当的能量利用和管理对于神经元进行处理并有效地交流。途径和ATP缓冲机制仍有争议。遗传编码的生物传感器的寿命成像，并准确地量化了急性bra刺刺激性刺激的实时MET-IND动力学颗粒神经元。 ATP Buffere神经元如何受到肌酸激酶或腺苷酸激酶同工酶的敲除。对生酮饮食的代谢改变的影响 - 一种非常低的碳水化合物，高脂饮食 - 癫痫的治疗方法将确定神经退行性疾病的基本病理学机制。在神经巡回赛上，显微镜，数据分析的生物信息学，科学写作，实验室领导和其他许多人都可以学习相对，并收到有关其数据和假设的反馈。培训经验他们过渡到职业生涯的下一阶段。