Characterization of Lphn3 function using a mutant murine model

使用突变小鼠模型表征 Lphn3 功能

基本信息

批准号：
8460516
负责人：
Deeann Wallis-Schultz
金额：
$ 15.82万
依托单位：
TEXAS A&M UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-20 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8460516
关键词：
Adult Affect Anxiety Attention Attention deficit hyperactivity disorder Behavior Behavior assessment Behavioral Behavioral Assay Biological Assay Brain Brain region Candidate Disease Gene Cell Differentiation process Clinical Consumption Data Development Diagnosis Disease Dopamine Receptor Etiology G Protein-Coupled Receptor Genes Gene Expression Genes Genetic Polymorphism Genotype Glutamates Growth Image Impulsivity In Vitro Knockout Mice Learning Measures Memory Mental Depression Modeling Molecular Analysis Motivation Motor Motor Activity Mus Mutant Strains Mice Neurites Neuronal Differentiation Neurons Orphan Pathway interactions Pattern Pharmaceutical Preparations Pharmacogenetics Phenotype Play Reaction Time Research Reverse Transcriptase Polymerase Chain Reaction Role Seminal Series Serotonin Serotonin Receptor 5-HT2A Signal Transduction Sucrose Swimming System Testing Time Tissue-Specific Gene Expression addiction base design dopaminergic neuron gene function genome-wide linkage grasp human GPRC5C protein inattention male monoamine mutant neuron development neurotransmitter release novel receptor response serotonin receptor serotonin transporter

项目摘要

DESCRIPTION (provided by applicant): Though relatively unknown and uncharacterized, the orphan GPCR LPHN3 has already been correlated in genome-wide linkage and association studies to both addictive phenotypes and Attention Deficit Hyperactivity Disorder (ADHD) independently. Vulnerability to addiction and ADHD are frequently comorbid, suggesting a common etiology. Further, initial data indicate that LPHN3 genotype may be a pharmacogenetic marker predicting response to stimulant medication. Unfortunately, as no disease-associated functional polymorphisms have been identified in LPHN3, its mechanism of action is completely unknown, and further characterization of its function is essential as there are far reaching implications for such an important gene. The central hypothesis to be tested in this proposal is that Lphn3 signalling affects neurotransmitter release and concentrations of monoamines in the brain that may affect neural cell differentiation, development, and function. These alterations ultimately play out in an effect on basic behavior that may influence phenotypes such as ADHD and vulnerability to addiction. We propose to utilize the novel Lphn3 null mutant mouse to investigate gene function. Preliminary data indicate that the Lphn3 null mouse is hyperactive, has altered DA and 5-HT levels in the brain, and demonstrates altered expression levels of genes involved in neuronal differentiation and development as well as other ADHD candidate genes involving the dopaminergic and serotonergic pathways including dopamine and serotonin receptors. These scant clues have been used by us to construct our research plan. We will begin with defining the Lphn3 developmental expression pattern in the brain and look at co-expression with other well-characterized neuronal markers. We will also utilize TaqMan Gene Expression Assays to look for changes in expression of genes known to play a role in neuronal differentiation, development, survival, and function as a function of Lphn3 genotype. Assays will be preformed across developmental time points and in specific brain regions. Next we will isolate primary neurons from WT and mutant mice and evaluate their viability, growth and function based on Lphn3 genotype. Finally, we have designed a battery of behavioral assays to begin to evaluate the basic behavioral deficits in the Lphn3 mice. This battery includes preliminary assessment of: activity, motor function, anxiety, depression, motivation, attention, impulsivity, learning and memory. Data derived from this project will be a first step toward characterizing the basic functions of Lphn3, which could enable better understanding, diagnosis, and treatment of ADHD and addiction.

描述（由申请人提供）：尽管相对未知和未表征，但孤儿GPCR LPHN3已经在全基因组联系和关联研究中与成瘾性表型和注意力缺陷多功能障碍（ADHD）相关。对成瘾和多动症的脆弱性通常是合并的，这表明是一种常见的病因。此外，初始数据表明LPHN3基因型可能是预测刺激药物反应的药物遗传学标记。不幸的是，由于在LPHN3中没有发现与疾病相关的功能多态性，因此其作用机理是完全未知的，并且其功能的进一步表征至关重要，因为对这种重要基因的影响很大。该提案中要检验的中心假设是LPHN3信号会影响神经递质释放和大脑中可能影响神经细胞分化，发育和功能的单胺浓度。这些改变最终会影响基本行为，可能会影响ADHD和成瘾脆弱性等表型。我们建议利用新型的LPHN3空突变小鼠研究基因功能。初步数据表明，LPHN3无效小鼠是多动，已经改变了大脑的DA和5-HT水平，并证明了参与神经元分化和发育的基因的表达水平改变了，以及其他ADHD候选基因，涉及多巴胺能和血清素途径，包括多巴胺和血清素受体。这些很少的线索已被我们构建我们的研究计划。我们将首先定义大脑中的LPHN3发育表达模式，并与其他特征良好的神经元标记共表达。我们还将利用Taqman基因表达分析来寻找已知在神经元分化，发育，生存和功能中发挥作用的基因表达变化，这是LPHN3基因型的函数。测定将在发育时间点和特定的大脑区域中预先形成。接下来，我们将从WT和突变小鼠中隔离原发性神经元，并根据LPHN3基因型评估其生存力，生长和功能。最后，我们设计了一系列的行为测定，以开始评估LPHN3小鼠的基本行为缺陷。该电池包括：活动，运动功能，焦虑，抑郁，动机，注意力，冲动，学习和记忆的初步评估。从该项目得出的数据将是表征LPHN3的基本功能的第一步，这可以使您可以更好地理解，诊断和治疗多动症和成瘾。