Recombinant Immunolabels for Nanoprecise Brain Mapping Across Scales

用于跨尺度纳米精确脑图谱的重组免疫标记

• 批准号: 10454277
• 负责人: James S Trimmer
• 金额: $ 140.12万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454277
• 关键词: Affinity; Antibodies; Archives; Array tomography; BRAIN initiative; Binding; Biological Assay; Brain; Brain Mapping; Cells; Collection; Cryopreservation; DNA Sequence; Effectiveness; Engineering; Ensure; Freezing; Functional disorder; Future; Goals; High-Throughput Nucleotide Sequencing; Human; Hybridomas; Image; Image Enhancement; Imaging Techniques; Individual; Label; Lead; Maps; Methods; Microscopy; Molecular; Monoclonal Antibodies; Mus; Neurosciences; Neurosciences Research; Penetration; Plasmids; Proteins; Proxy; Reagent; Recombinant Antibody; Recombinants; Reproducibility; Research; Research Personnel; Resolution; Resources; Sampling; Structure; Subcellular structure; Synapses; System; Techniques; Tissues; Validation; Work; affinity labeling; anatomical tracing; antibody engineering; base; brain circuitry; brain tissue; cell type; cost; immunoreactivity; improved; interest; miniaturize; nanobodies; nanometer; nanoscale; nonhuman primate; novel; open source; plasmid DNA; preservation; repository; screening; single molecule

Recombinant Immunolabels for Nanoprecise Brain Mapping Across Scales Understanding brain function and dysfunction requires an understanding of the circuitry of the brain from molecules to cells to circuits. While no single technique can achieve this, a strategic combination of techniques applied across scales can provide information that when integrated can lead to a more complete picture. These techniques can range from analyses of single molecules, including their localization in nanodomains, to subcellular compartments such as synapses and synaptic networks to entire brains. One common theme of these techniques is that they require affinity probes that specifically label subcellular structures, cell types, and circuits within the brain. We propose to enhance an existing resource of renewable affinity probes in the form of an extensive collection of highly-validated monoclonal antibodies. We will enhance our ongoing dissemination of low cost, high quality antibodies to neuroscience researchers by converting these to recombinant form. This will also ensure permanence of this valuable collection of renewable reagents for future researchers. We will use established methods to miniaturize these conventional antibodies into a nanoscale form. These miniaturized antibodies will allow for labeling of brain targets with single nanometer precision, which will provide more spatially precise labeling and overcome the limits to imaging resolution that conventional Abs represent. Moreover, the miniaturized Abs will have much better sample penetration, allowing for more efficient labeling of larger samples of the type used for defining intact circuits. We have formed a consortium of experts who developed and/or are experts in a powerful set of advanced techniques that together allow for brain mapping across scales. This consortium will validate nanoscale antibodies in their respective techniques, a key component of rigor and reproducibility of any antibody-based research. We will also generate and validate novel nanoscale antibodies against targets of great interest to brain investigators for which no suitable reagents exist. Together, these efforts will further enhance the dissemination of this valuable resource, and fundamentally accelerate the pace of brain circuit mapping across scales.

重组免疫标记物用于跨尺度的纳米级脑成像 要了解大脑的功能和功能障碍，需要了解大脑的回路 从分子到细胞再到电路。虽然没有一种单一的技术可以实现这一点，但技术的战略组合 跨规模应用可以提供信息，当这些信息整合在一起时，可以产生更完整的画面。这些 技术的范围可以从单分子的分析，包括它们在纳米域中的定位，到 亚细胞室，如突触和突触网络，通向整个大脑。 这些技术的一个共同主题是，它们需要亲和探针来专门标记亚细胞 大脑内部的结构、细胞类型和回路。我们建议加强现有的可再生能源资源 亲和力探针的形式广泛收集高度验证的单抗。我们将加强 我们正在向神经科学研究人员传播低成本、高质量的抗体，通过将这些 变成重组的形式。这也将确保这一宝贵的可再生试剂收藏的永久性 未来的研究人员。我们将使用现有的方法将这些常规抗体微型化为 纳米级的形状。这些微型化的抗体将允许用单个纳米标记大脑靶标 精度，这将提供更精确的空间标记，并克服成像分辨率的限制 常规抗体代表了。此外，小型化的抗体将具有更好的样品渗透性，从而 以便更有效地标记用于定义完整电路的类型的较大样本。 我们已经组成了一个专家联盟，他们开发和/或是一套强大的先进的 这些技术结合在一起，可以在不同的范围内绘制大脑图谱。该联盟将验证纳米级 抗体在各自的技术中，是任何基于抗体的严密性和重复性的关键组成部分 研究。我们还将产生和验证针对大脑非常感兴趣的目标的新型纳米级抗体。 没有合适试剂的调查人员。这些努力一起，将进一步加强传播。 这一宝贵的资源，并从根本上加快了跨尺度的大脑回路映射的步伐。