Analytical Methods for Investigating Peptide Transport

研究肽转运的分析方法

基本信息

批准号：
8092548
负责人：
Susan M Lunte
金额：
$ 34.23万
依托单位：
UNIVERSITY OF KANSAS LAWRENCE
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-03-18 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8092548
关键词：
Adverse effects Affect Agonist Alzheimer&apos s Disease Amides Amino Acid Neurotransmitters Amino Acid Sequence Analgesics Animals Area Astrocytes Attention Autopsy Back Behavior Behavior monitoring Binding Biological Blood - brain barrier anatomy Brain C-terminal Catecholamines Cattle Cells Clinical Cocaine Dependence Coculture Techniques Codeine Detection Development Disease Dopamine Drug Addiction Drug Industry Dynorphin A Dynorphins Electrophoresis Endothelial Cells Excitatory Amino Acids Exhibits Extracellular Space Glutamates Glutamine Goals Grant Health Hormones In Vitro Investigation Lead Ligands Limb structure Marketing Measurement Mediating Mental Depression Metabolism Methodology Microchip Electrophoresis Microdialysis Monitor Mood Disorders Morphine N-Methyl-D-Aspartate Receptors N2-tryptophyllysine Neuromodulator Neurons Neuropeptides Neuropharmacology Neurotransmitters Nitric Oxide Organism Pain Paralysed Parents Patients Peptide Fragments Peptide Metabolism Peptide Transport Peptides Peripheral Pharmaceutical Preparations Physiological Play Process Property Rattus Reporting Research Role Sampling Scientist Sheep Site Substance P Symptoms System Techniques Telemetry Therapeutic Therapeutic Effect Tissues addiction analog analytical method arginylarginine arginylproline awake base in vivo interest kappa opioid receptors leucylarginine leucylmethionine microchip mu opioid receptors neurochemistry neurotoxic neurotransmitter release painful neuropathy phenylalanyl-phenylalanyl-glycine phenylalanylleucine postin pre-clinical receptor research study rimorphin sensor sensory stimulus small molecule therapeutic target tyrosyl-glycyl-glycyl-phenylalanyl

项目摘要

DESCRIPTION (provided by applicant): Neuropeptides play multiple roles in vivo including that of a neurotransmitter, neuromodulator, and hormone. In contrast to small molecule neurotransmitters such as glutamate and dopamine, neuropeptides are not taken back up by neurons following their release but are metabolized in the extracellular space into smaller peptide fragments. These fragments can directly affect the cells in the extracellular space in the surrounding area, pass through the blood brain barrier (BBB), and exert effects at sites far removed from the site of their initial release. In this proposal, the transport, metabolism and neuropharmacology of dynorphin A (Dyn A) is investigated both in vivo and in vitro using LC-MS and on-line microdialysis-microchip electrophoresis. Dyn A is a natural ligand of the kappa opioid receptor. Analogs of dynorphin and its metabolites have been investigated for the treatment of peripheral pain, drug addiction, and affective disorders. Paradoxically, Dyn A has also been shown to be neurotoxic. An elevated level of this peptide in the CNS has been associated with neuropathic pain and Alzheimer's disease. There are three main goals of this proposal. The first is to investigate the transport and metabolism of Dyn A 1-17, its metabolites, and synthetic analogs across the BBB. Information acquired through these experiments will lead to a better understanding of the mechanism of transport and will make it possible to produce better therapeutics targeted at the kappa opioid receptor. The second is to develop separation-based sensors that can be used to monitor the effects of Dyn A and its metabolites on neurotransmitter release and neuropeptides in the CNS. The third objective is to develop a separation-based sensor that can be placed on-animal so that measurements of neurotransmitters and behavior can be made simultaneously, using awake, freely roaming animals. These techniques will be employed to study the pharmacological actions of Dyn A at the BBB and in the CNS. The methodology developed here will be generally applicable to all neurochemical studies in which the measurement of blood brain barrier transport or neurotransmitters is desired, especially those cases where monitoring behavior is also of interest. PUBLIC HEALTH RELEVANCE Dynorphin is an important neuropeptide that is involved in pain, cocaine addiction, depression, and Alzheimer's disease. A better understanding of the transport and metabolism of this peptide across the BBB will assist scientists in producing better treatments for these diseases. The analytical methods developed in this proposal will also be of general utility to scientists involved in CNS research.

描述（由申请人提供）：神经肽在体内扮演多个角色，包括神经递质，神经调节剂和激素。与小分子神经递质（如谷氨酸和多巴胺）相反，神经肽在释放后不会被神经元备份，而是在细胞外空间中代谢成较小的肽片段。这些碎片可以直接影响周围地区细胞外空间中的细胞，穿过血脑屏障（BBB），并在远离其最初释放位置的位置施加效果。在此提案中，使用LC-MS和在线微透析 - 微观芯片电泳中研究了Dynorphin A（Dyno A）的转运，代谢和神经药理。 Dyn A是Kappa阿片受体的天然配体。已经研究了测能力及其代谢产物的类似物，以治疗外周疼痛，药物成瘾和情感障碍。矛盾的是，Dyn A也已被证明是神经毒性的。中枢神经系统中这种肽的水平升高与神经性疼痛和阿尔茨海默氏病有关。该提案有三个主要目标。首先是研究Dyn A 1-17的运输和代谢，其代谢产物和整个BBB的合成类似物。通过这些实验获得的信息将使人们更好地理解运输机制，并将有可能产生针对Kappa阿片类药物受体的更好的治疗剂。第二个是开发基于分离的传感器，这些传感器可用于监测Dyn A及其代谢产物对CNS中神经递质释放和神经肽的影响。第三个目标是开发一个基于分离的传感器，该传感器可以放置在动物上，以便可以使用清醒的，自由漫游的动物同时进行神经递质和行为的测量。这些技术将用于研究BBB和中枢神经系统中Dyn A的药理作用。此处开发的方法通常适用于所有神经化学研究，其中需要测量血液屏障转运或神经递质，尤其是那些也引起监测行为的情况。公共卫生相关性驱虫酚是一种重要的神经肽，与疼痛，可卡因成瘾，抑郁症和阿尔茨海默氏病有关。更好地了解该肽在BBB的运输和代谢将有助于科学家为这些疾病提供更好的治疗方法。该提案中开发的分析方法也将对参与中枢神经系统研究的科学家提供一般效用。