Genetic and Optogenetic Models to Dissect the Role of the Serotonin 1B Receptor

遗传和光遗传学模型剖析 5-羟色胺 1B 受体的作用

基本信息

批准号：
8685028
负责人：
Katherine M Nautiyal
金额：
$ 5.51万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8685028
关键词：
A Mouse Actins Address Adult Aggressive behavior Attention Attention deficit hyperactivity disorder Autoradiography Autoreceptors Basal Ganglia Behavior Behavioral Behavioral Assay Bipolar Disorder Brain Brain region Cells Cerebellum Code Coupled Development Disease Doxycycline Fiber Optics G Protein-Coupled Receptor Genes Generations Genes Genetic Genetic Polymorphism Goals Halorhodopsins Hippocampus (Brain)Human Hyperactive behavior Hypothalamic structure Implant Impulsive Behavior Impulsivity In Situ Hybridization Injection of therapeutic agent Intervention Knock-out Knockout Mice Light Measures Mediating Mediation Mental Depression Mental disorders Modeling Mus Nerve Neurons Pattern Pharmacology Phenotype Play Population Predisposition Presynaptic Terminals Prosencephalon Pump Regulation Research Role Schizophrenia Serotonin Serotonin Receptor 5-HT1B Signal Transduction Suicide Symptoms Synapses System Tetracyclines Thalamic structure Time Tissues Transgenes Transgenic Mice Violence Viral Vector addiction calmodulin-dependent protein kinase II critical period follow-up knock-down mouse model neural circuit neuropsychiatry neurotransmitter release neurotrophic factor novel optogenetics programs promoter public health relevance receptor receptor binding serotonin receptor suicidal tool

项目摘要

DESCRIPTION (provided by applicant): Aggression and impulsivity are two understudied aspects of behavior that are significant symptoms in a number of psychiatric disorders. The serotonin 1B receptor (5-HT1B R) has been implicated in aggressive and impulsive behavior using pharmacology, constitutive knock-out mouse models, and human gene polymorphism association studies. Given evidence of serotonergic modulation of brain development, it is likely that 5-HT1B R plays a developmental role in establishing circuits underlying aggressive and impulsive behavior. 5-HT1B R is a GPCR that inhibits neurotransmitter release in axon terminals of serotonergic and non-serotonergic neurons localized broadly in the basal ganglia, hippocampus, thalamus, cerebellum and raphe. Thus, an inducible and tissue specific strategy is necessary to delineate the sensitive period and neural circuits through which 5-HT1B Rs regulate aggression and impulsivity. We have therefore generated a mouse line that permits temporal and spatial regulation of 5-HT1B R. A tetracycline operator (tetO) has been inserted between the promoter and coding region of the 5-HT1B R (tetO1B). Receptor binding and in situ hybridization reveals that these tetO1B mice have normal basal 5-HT1B R expression in the brain compared to 129 WT mice. Crossing tetO1B mice to transgenic mice expressing the tetracycline-dependent transcriptional silencer (tTS) under the control of ¿-Actin (ubiquitous), CaMKII (forebrain-specific) or Pet (raphe-specific) promoters yields mouse lines with tissue specific and inducible knock-down of 5-HT1B R. Initial characterization with autoradiography and in situ hybridization reveals promoter-specific 5-HT1B R expression patterns that can be modulated with doxycycline administration. Preliminary behavioral assays using tetO1B:: ¿-Actin-tTS+ mice reveal a similar behavioral phenotype to the constitutive knock-out mouse-i.e., significantly increased impulsive aggression. Therefore, we have the tools to manipulate the expression of 5-HT1B R spatially and temporally and examine the resulting impact on aggressive and impulsive behaviors. Follow-up studies using optogenetics will address the circuit level mechanisms by determining which brain regions in the aggression and impulsivity circuitry in adulthood may have been altered by the intervention during the critical period. The hypothesis is that a lack of 5HT1B Rs during a critical period of development results in an overactive 'aggression circuit', and so by inhibiting these overactive brain regions, a normal phenotype can be rescued. This is possible using the light-sensitive inhibitory Cl- pump, halorhodopsin, introduced through viral vector injection into target brain regions, with subsequent activation by yellow light administered via fiber optic implant. Overall, understanding how 5-HT1B R mediates the neural circuits underlying aggression and impulsivity will have implications for the treatments of several neuropsychiatric disorders, including addiction, attention-deficit hyperactivity disorde and bipolar disorder, and suicidal depression.

描述（通过应用程序提供）：侵略性和冲动性是行为的两个知识，是许多精神疾病中的重要症状。羟色胺1B受体（5-HT1B R）使用药理学，构成敲除小鼠模型和人类基因多态性关联研究与攻击性和冲动行为有关。鉴于脑发育的血清素能调节的证据，5-HT1B R可能在建立侵略性和冲动行为的基础电路中起发育作用。 5-HT1B R是一种GPCR，可抑制在塞拉顿能和非杂质神经元的轴突末端释放神经递质的释放，该神经元在基底神经节，海马，thalamus，cerebellum和raphe中广泛定位。这是描绘敏感时期和神经元的诱导和组织特定策略，其中5-HT1B RS调节攻击性和冲动性。因此，我们生成了一条允许5-HT1B的临时和空间调节的小鼠线。四环素算子（TETO）已在5-HT1B R（TETO1B）的启动子和编码区域之间插入。受体结合和原位杂交表明，与129 wt小鼠相比，这些TETO1B小鼠的基础5-HT1B R表达正常。在表达四环素依赖性转录消音器（TTS）的转基因小鼠的跨基因小鼠（tts）下，在控制�-肌动蛋白（无处不在），caMKII（前脑特异性）或PET（Raphe特异性）启动子（Raphe特异性）启动子中产生的小鼠系和诱导的小鼠构图R.最初的5-htrbrization R.揭示了可以通过强力霉素给药调节的启动子特异性的5-HT1B R表达模式。使用TETO1B ::¿-Actin-TTS+小鼠的初步行为测定显示与本构敲除小鼠I.e。相似的行为表型，显着提高了脉冲性侵略性。因此，我们有工具可以在空间和临时操纵5-HT1B R的表达，并检查对侵略性和冲动行为的影响。使用光遗传学的后续研究将通过确定在关键时期的干预措施可能改变了成年后的侵略性和冲动电路中的哪些大脑区域来解决电路水平机制。假设是，在关键发展期间缺乏5HT1B RS会导致过度活跃的“攻击回路”，因此通过抑制这些过度活跃的大脑区域，可以挽救正常的表型。使用光敏感性抑制性克服，通过病毒载体注入到靶脑区域中，随后通过纤维植入物给予的黄光激活，这是可能的。总体而言，了解5-HT1B R如何介导侵略性和冲动性的神经元电路将对治疗产生影响在几种神经精神疾病中，包括加法，注意力缺陷多动障碍和躁郁症以及自杀抑郁症。