Effect of neurotensin analogs on protein phosphorylation in the mouse brain

神经降压素类似物对小鼠脑内蛋白质磷酸化的影响

基本信息

批准号：
8053376
负责人：
MONA M BOULES
金额：
$ 7.65万
依托单位：
MAYO CLINIC JACKSONVILLE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8053376
关键词：
Acute Adverse effects Affinity Agonist Amino Acids Animal Model Animals Antipsychotic Agents Binding Blood - brain barrier anatomy Brain Brain region Cell physiology Clozapine Corpus striatum structure Data Detection Development Disease Dissection Dopamine Drug usage Effectiveness Event Exhibits Functional disorder Goals Haloperidol Hypotension Inbred C57BL Mice Injection of therapeutic agent Knockout Mice Knowledge Laboratories Mass Spectrum Analysis Mediating Methods Molecular Target Mus Neuropeptides Neurotensin Neurotensin Receptors Neurotransmitters Nucleus Accumbens Pain Peptide Hydrolases Phosphoproteins Phosphorylation Play Preparation Property Proteins Research Role Schizophrenia Signal Transduction Synapses System Testing Therapeutic Time Two-Dimensional Gel Electrophoresis Wild Type Mouse analog atypical antipsychotic design frontal lobe genetic regulatory protein insight natural hypothermia neuropsychiatry neurotensin 69L novel public health relevance receptor

项目摘要

DESCRIPTION (provided by applicant): The goals of the proposed research are to 1) compare the effects on signal transduction of specific neurotensin (NT) analogs, NT69L and NT79, to those of typical and atypical antipsychotics, and 2) to identify the NT receptors (NTRs) by which these effects are mediated. The long term objective of the proposed research is to better understand the mechanism(s) of action of NT analogs to further their development as potential novel antipsychotic drugs. With this knowledge, NT analogs can be modified to directly target proteins thought to be directly involved in the treatment of neuropsychiatric disease while avoiding those thought to mediate adverse side effects. Neurotensin (NT) is an endogenous thirteen amino acid neuropeptide that acts as a neurotransmitter in the brain. It has been implicated in the pathophysiology of neuropsychiatric diseases including schizophrenia, as well as in the mechanism of action of antipsychotic drugs. NT can elicit effects similar to those seen using antipsychotic drugs, even when administered acutely. However, in animal models NT must be administered directly into the brain to observe its effects. To circumvent this problem, the Richelson laboratory has developed several NT analogs that act as agonists to one or more of the three known NT receptors, NTS1, NTS2, and NTS3. The NT analogs have similar effects as NT, but their mechanisms of action are not fully understood at this time. These analogs, which can be administered peripherally, include NT69L, thought to bind both NTS1 and NTS2, and NT79, thought to preferentially bind NTS2. Comparing the effects of these compounds on protein phosphorylation in regions of the brain implicated in schizophrenia to the effects of established antipsychotic drugs will offer insight into the mechanism(s) of action mediating antipsychotic-like effects. This will further the development of NT analogs as potential antipsychotic drugs by identifying possible molecular targets which mediate the antipsychotic-like properties of the analogs as well as identifying potential adverse side-effects. Three focused research aims are proposed to elucidate the mechanism(s) of action of these analogs: (1) Identify proteins that are differentially phosphorylated in wild-type C57BL mice treated acutely with vehicle, NT69L, and NT79, and compare them to those treated acutely with haloperidol and clozapine, (2) Identify proteins that are differentially phosphorylated between NTS1 knock-out (KO) mice treated acutely with vehicle, NT69L and NT79, and (3) Identify proteins that are differentially phosphorylated between NTS2 KO mice treated acutely with vehicle, NT69L and NT79. Methods include animal injection, brain dissection, extraction of synaptoneurosomal-enriched preparations, and protein separation via two-dimensional gel electrophoresis, phosphoprotein detection and identification of differentially phosphorylated proteins via mass spectrometry. PUBLIC HEALTH RELEVANCE: NT69L and NT79 are compounds that may be therapeutic in the treatment of schizophrenia. Determining the effects of these compounds on proteins in the brain, and comparing them to those caused by established antipsychotic drugs (used to treat schizophrenia), can offer insight to what role each compound may play in treating disease. With this knowledge, these compounds may be designed to more specifically target those proteins involved in disease, increasing their effectiveness as antipsychotics and decreasing potential side effects.

描述（由申请人提供）：拟议的研究的目标是1）比较对特定神经素（NT）类似物（NT69L和NT79）信号转导的影响与典型和非典型抗精神病药的影响，以及2）这些作用介导的受体（NTR）。拟议的研究的长期目标是更好地了解NT类似物的作用机制，以进一步发展其作为潜在的新型抗精神病药。有了这些知识，可以对NT类似物进行修改，以直接靶向被认为直接参与神经精神疾病的蛋白质，同时避免了那些被认为介导不良副作用的蛋白质。神经素（NT）是一种内源性13个氨基酸神经肽，在大脑中充当神经递质。它与包括精神分裂症在内的神经精神疾病的病理生理以及抗精神病药作用机理有关。 NT可以引起类似于使用抗精神病药的效果，即使急性施用也是如此。但是，在动物模型中，必须直接将NT给药以观察其作用。为了解决这个问题，Richelson实验室已经开发了几种NT类似物，这些类似物是对三个已知NT受体NTS1，NTS2和NTS3中一个或多个激动剂的激动剂。 NT类似物具有与NT相似的效果，但目前尚未完全理解其作用机理。这些类似物可以在外围施用，包括NT69L，被认为既结合NTS1和NTS2和NT79），被认为优先结合NTS2。将这些化合物对与精神分裂症有关的大脑区域中蛋白质磷酸化的影响与已建立的抗精神病药的作用相比，将洞悉介导抗精神病药的作用机制。这将进一步发展NT类似物作为潜在的抗精神病药，通过鉴定可能介导类似物的抗精神病药样特性并确定潜在的不良副作用的可能分子靶标。提出了三个重点研究的目的，以阐明这些类似物的作用机制：（1）鉴定在野生型C57BL小鼠中差异化磷酸化的蛋白质，并用媒介物，NT69L和NT79急性磷酸（2）在NTS1敲除（KO）小鼠之间急性地鉴定蛋白质，用媒介物，NT69L和NT79急性磷酸化，（KO）小鼠，（3）蛋白质在NTS2 KO小鼠之间差异化的蛋白质，车辆，NT69L和NT79。方法包括动物注射，脑解剖，突触核素添加的制剂的提取以及通过二维凝胶电泳的蛋白质分离，磷酸蛋白检测以及通过质谱法鉴定差异磷酸化的蛋白质。公共卫生相关性：NT69L和NT79是在精神分裂症治疗中可能具有治疗性的化合物。确定这些化合物对大脑蛋白质的作用，并将其与由已建立的抗精神病药（用于治疗精神分裂症）引起的蛋白质进行比较，可以洞悉每种化合物在治疗疾病中的作用。有了这些知识，这些化合物可以设计为更具体地靶向参与疾病的蛋白质，从而提高其作为抗精神病药的有效性并降低潜在的副作用。