MicroRNA101 and the Termination of Early Phase Neural Development

MicroRNA101 与早期神经发育的终止

• 批准号: 8914061
• 负责人: Darwin K BERG
• 金额: $ 19.38万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8914061
• 关键词: AMPA Receptors; Accounting; Acute; Address; Adult; Amyloid; Automobile Driving; Biological Neural Networks; Brain; Calcium; Chloride Ion; Chlorides; Complex; Development; Epilepsy; Event; Failure; Frequencies; Future; Goals; Health; Image; Individual; Intervention; Investigation; Life; Mediating; Messenger RNA; MicroRNAs; N-Methyl-D-Aspartate Receptors; Nervous system structure; Neuronal Differentiation; Neurons; Outcome; Pathway interactions; Phase; Play; Population; Porifera; Process; Proteins; Regulation; Regulatory Pathway; Reporting; Role; Schedule; Shapes; Signal Transduction; Site; Slice; Synapses; Synaptic plasticity; System; Testing; Therapeutic; Time; Work; cohort; gamma-Aminobutyric Acid; in vivo; inhibitor/antagonist; insight; interest; nervous system development; nervous system disorder; neurodevelopment; neuronal growth; patch clamp; postnatal; prevent; regenerative; synaptogenesis; transmission process

DESCRIPTION (provided by applicant): The developing nervous system undergoes a major transition in early postnatal life when it shifts from a period of exuberant synapse formation to a subsequent period of synaptic pruning and network consolidation. Synaptic numbers approach a maximum during this time, the rules governing synaptic plasticity change, AMPA and NMDA receptor compositions are altered, and GABAergic signaling shifts from being depolarizing/excitatory to hyperpolarizing/inhibitory, in addition to other changes. Mechanisms driving this profound transition are poorly understood at best. Interesting candidates for doing so are microRNAs (miRs) because they can target many different mRNAs at the same time for blockade or destruction, thereby producing a "regulatory hub" to coordinate complex changes across large systems. Our preliminary evidence indicates that miR-101 is an attractive candidate for executing this transition. It is abundant, increases substantially at the relevant time, and remains high in the adult. Using antagonists to block miR-101 function in vivo reveals significant aberrations: substantial increases are seen in spontaneous synchronized activity across neuronal populations and is accompanied by increases in synaptic number and excitatory synaptic input to neurons. Blocking miR-101 also appears to delay maturation of GABAergic signaling, enabling it to be depolarizing at later times. Importantly, our preliminary results using target site blockers to protect specific mRNAs suggest that miR-101 achieves its effects by acting on multiple targets. One is likely to be NKCC1, the chloride transporter responsible for GABA being depolarizing. But other miR-101 targets appear to be important as well, suggesting that terminating the depolarizing phase of GABAergic signaling is not by itself sufficient to account for all the major changes comprising the developmental transition. We will test the role of miR-101 in mediating the transition in nervous system development by using antagonists, sponges, and target site blockers to prevent its action in vivo while assessing the consequences for synaptic activity and network function. Calcium fluors in acute slices will report the frequency and extent of spontaneous coordinated activity across neuronal populations, as well as total activity and numbers of participating neurons. Patch-clamp recording will reveal type and amount of synaptic activity while immunostaining will quantify synapses. By selecting individual miR-101 targets for protection, it will be possible to dissect th contributions of different pathways to the transition and to evaluate the consequences of defective regulation. This will provide new insight into mechanisms driving the transition, reveal the role of miR-101 in particular, and likely have biomedical relevance by revealing the vulnerability of the system to individual regulatory pathways being compromised, producing, for example, greater propensity for epileptic seizure-like events.

描述（由申请人提供）：发育中的神经系统在出生后的早期经历了一个主要的转变，当时它从旺盛的突触形成期转变为 随后突触修剪和网络巩固时期。在此期间，突触数量接近最大值，控制突触可塑性的规则发生变化，AMPA和NMDA受体组成发生变化，GABA能信号从去极化/兴奋转变为超极化/抑制，以及其他变化。推动这一深刻转变的机制充其量也只是知之甚少。有趣的候选人这样做 是微小RNA（miR），因为它们可以同时靶向许多不同的mRNA进行阻断或破坏，从而产生一个“调节中心”来协调大型系统中的复杂变化。我们的初步证据表明，miR-101是执行这种转变的有吸引力的候选者。它是丰富的，在相关的时间大幅增加，并保持在成人高。使用拮抗剂阻断体内miR-101功能揭示了显著的畸变：在神经元群体中观察到自发同步活动的显著增加，并伴随着突触数量和神经元兴奋性突触输入的增加。阻断miR-101似乎也延迟了GABA能信号传导的成熟，使其能够在稍后时间去极化。重要的是，我们使用靶点阻断剂保护特定mRNA的初步结果表明，miR-101通过作用于多个靶点来实现其作用。一个可能是NKCC 1，负责GABA去极化的氯转运蛋白。但其他miR-101靶点似乎也很重要，这表明终止GABA能信号的去极化阶段本身不足以解释发育过渡中的所有主要变化。我们将测试miR-101在神经系统发育中介导的过渡中的作用，通过使用拮抗剂，海绵和靶点阻断剂来防止其在体内的作用，同时评估突触活性和网络功能的后果。急性切片中的钙荧光剂将报告神经元群体之间自发协调活动的频率和程度，以及参与神经元的总活动和数量。膜片钳记录将揭示突触活动的类型和数量，而免疫染色将量化突触。通过选择单个miR-101保护靶点，将有可能分析不同途径对过渡的贡献，并评估缺陷调控的后果。这将为驱动这种转变的机制提供新的见解，特别是揭示miR-101的作用，并可能通过揭示系统对个体调控途径的脆弱性而具有生物医学相关性，例如，产生更大的癫痫样事件倾向。

项目成果

MicroRNA-101 Regulates Multiple Developmental Programs to Constrain Excitation in Adult Neural Networks.

• DOI: 10.1016/j.neuron.2016.11.017
• 发表时间: 2016-12-21
• 期刊: Neuron
• 影响因子: 16.2
• 作者: Lippi G;Fernandes CC;Ewell LA;John D;Romoli B;Curia G;Taylor SR;Frady EP;Jensen AB;Liu JC;Chaabane MM;Belal C;Nathanson JL;Zoli M;Leutgeb JK;Biagini G;Yeo GW;Berg DK
• 通讯作者: Berg DK