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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

基本信息

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
项目类别：
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

项目摘要

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.

用于跨尺度纳米精确脑标测的重组免疫标记了解大脑功能和功能障碍需要了解大脑的电路，分子到细胞到电路虽然没有单一的技术可以做到这一点，一个战略性的技术组合，跨尺度应用可提供信息，综合这些信息后可得出更全面的情况。这些技术的范围可以从单个分子的分析，包括它们在纳米结构域中的定位，亚细胞区室，如突触和突触网络到整个大脑。这些技术的一个共同主题是，它们需要特异性标记亚细胞的亲和探针，结构，细胞类型和大脑内的电路。我们建议加强现有的可再生能源，亲和探针以高度验证的单克隆抗体的广泛集合的形式。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，重组形式。这也将确保这些宝贵的可再生试剂的永久性，未来的研究者我们将使用已建立的方法将这些常规抗体纯化成 nanoscale形式。这些微型化的抗体将允许用单个纳米标记大脑靶点。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，传统的ABS代表。此外，小型化的Ab将具有更好的样品渗透，用于更有效地标记用于定义完整电路的类型的较大样本。我们已经成立了一个专家联盟，他们开发了一套强大的先进的这些技术一起允许跨尺度的大脑映射。该联盟将验证纳米级抗体在其各自的技术，一个关键组成部分的严谨性和再现性的任何抗体为基础的 research.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抗体没有合适试剂的研究者。这些努力将进一步加强传播这一宝贵的资源，并从根本上加快跨尺度脑回路映射的步伐。