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In Vivo Brain Dialysis of Neuropeptides and Neuroimmune Signaling Proteins

神经肽和神经免疫信号蛋白的体内脑透析

基本信息

批准号：
8328931
负责人：
JULIE A STENKEN
金额：
$ 17.31万
依托单位：
UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-15 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8328931
关键词：
Acids Affect Affinity Alcoholism Amyloid Analytical Chemistry Animal Model Animals Antibodies Anxiety Biochemical Biochemistry Biological Assay Brain CCL2 gene CCL3 gene CCL4 gene Calibration Cells Centrifugation Chemicals Cholecystokinin Collection Communication Communities Complex Corticotropin-Releasing Hormone Coupled Cytokine Receptors Data Desire for food Dialysis procedure Disease Epilepsy Equilibrium Fever Hippocampus (Brain)Human Implant Infusion procedures Interleukin-10 Interleukin-6 Kainic Acid Letters Life Liquid substance Measurement Measures Medical Memory Mental Depression Mental disorders Messenger RNA Methods Microdialysis Microspheres Modeling Modification Molecular Weight Multiple Sclerosis Nervous system structure Neuraxis Neurons Neuropeptides Neuroregulator Neurosciences Neurosciences Research Neurotransmitters Pain Peptides Perfusion Procedures Process Proteins Pump RANTES Rattus Recovery Recycling Relative (related person)Research Risk Rodent Rodent Model Sampling Signal Transduction Signaling Protein Site Sleep Small Inducible Cytokine A3 System TNF gene Time Tissues Work analytical method angiotensin II, des-Asp(1)-des-Arg(2)-Ile(5)-base chemokine clinically significant cytokine extracellular fluid flow human disease improved in vivo innovation meetings nervous system disorder neuropeptide Y prevent receptor solute success tool uptake

项目摘要

DESCRIPTION (provided by applicant): Clinical Significance. Neuronal communication involves complex and coupled networks of different chemicals including neuropeptides, neurotransmitters, and signaling proteins called cytokines. Chemokines (chemoattractant cytokines) and cytokines act as immunoregulators and neuroregulators in the central nervous system (CNS) and are considered a third neurocommunication system. These important proteins affect many different disease states. A comprehensive understanding of the nervous system cannot be achieved without decoding this chemical signaling network. One of the primary barriers preventing decoding these chemical networks is the lack of appropriate chemical analysis tools that can be applied to study in vivo biochemistry in rodent models. In particular, microdialysis sampling of these important cytokines and peptides has been difficult to implement. Neuropeptide and cytokine ECF concentrations are generally unknown and their processing via receptor meditated uptake (cell-based receptors and for cytokines - soluble receptors) is poorly understood. The risks associated with performing this research are greatly offset by several high-impact benefits. 1) Significant biochemical information about the real protein concentrations and not just upregulated mRNA levels and their makeup (e.g., which cytokines) in the CNS will be gained. 2) True in vivo measurements will be made rather than post-mortem measurements. Experimental Approach. We hypothesize that modification to existing brain dialysis methods of either inclusion of a free antibody or antibodies immobilized to small microspheres will significantly improve protein relative recovery into dialysis probes that are implanted into the rat brain. Additionally, new flow regimes including recycled flow and flow reversal with new pumps are included as methods to increase relative recovery of the targeted proteins. Our primary focus will be on a select group of cytokines and neuropeptides related to epilepsy (cytokines {CCL2 [MCP-1], CCL3 [MIP-1a], CCL4 [MIP-12], CCL5 [RANTES], KC/GRO, IL-1b, IL-6, IL-10, and TNF-a} and neuropeptides {angiotensin IV, 2-amyloid, cholecystokinin, corticotropin releasing hormone, and neuropeptide Y). These multiplexed assays will allow for the simultaneous measurement of different peptides in volumes as low as 25 5L of dialysate collected from the rat brain.

描述（由申请人提供）：临床意义。神经元通讯涉及不同化学物质的复杂耦合网络，包括神经肽、神经递质和称为细胞因子的信号蛋白。趋化因子（化学引诱细胞因子）和细胞因子在中枢神经系统（CNS）中充当免疫调节剂和神经调节剂，被认为是第三神经通讯系统。这些重要的蛋白质影响许多不同的疾病状态。如果不解码这种化学信号网络，就无法全面了解神经系统。阻止解码这些化学网络的主要障碍之一是缺乏适当的化学分析工具，可以应用于研究啮齿动物模型中的体内生物化学。特别是，这些重要的细胞因子和肽的微透析取样一直难以实施。神经肽和细胞因子ECF浓度通常是未知的，并且它们通过受体介导的摄取（基于细胞的受体和对于细胞因子-可溶性受体）的加工知之甚少。与进行这项研究相关的风险被几个高影响力的好处大大抵消。 1)有关真实的蛋白质浓度的重要生化信息，而不仅仅是上调的mRNA水平及其组成（例如，将获得CNS中的细胞因子。 2)将进行真实的体内测量，而不是尸检测量。实验方法。我们假设，对现有的脑透析方法进行修改，包括游离抗体或固定在小微球上的抗体，将显著提高植入大鼠脑中的透析探针中的蛋白质相对回收率。此外，包括利用新泵的再循环流动和流动反转的新流动方案被包括作为增加靶向蛋白质的相对回收的方法。我们的主要重点将是与癫痫相关的一组选定的细胞因子和神经肽（细胞因子{CCL 2 [MCP-1]、CCL 3 [MIP-1a]、CCL 4 [MIP-12]、CCL 5 [RANTES]、KC/GRO、IL-1b、IL-6、IL-10和TNF-α}和神经肽{血管紧张素IV、2-淀粉样蛋白、胆囊收缩素、促肾上腺皮质激素释放激素和神经肽Y）。这些多重测定将允许同时测量不同的肽的体积低至25 - 5L的透析液从大鼠脑中收集。