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（由申请人提供）：攻击性和冲动性是行为的两个尚未充分研究的方面，是许多精神疾病的重要症状。使用药理学、本构性敲击，血清素 1B 受体 (5-HT1B R) 与攻击性和冲动行为有关。 -out 小鼠模型和人类基因多态性关联研究给出了大脑发育的血清素调节的证据，5-HT1B R 很可能是这样的。 5-HT1B R 是一种 GPCR，它在建立攻击性和冲动行为的基础电路中发挥着发育作用，可抑制广泛分布于基底神经节、海马、丘脑和中缝的血清素能和非血清素能神经元轴突末端的神经递质释放。，需要一种诱导性和组织特异性策略来描绘敏感期和神经回路，5-HT1B Rs 通过这些策略调节攻击性和因此，我们产生了允许 5-HT1B R 的时间和空间调节的小鼠品系。四环素操纵子 (tetO) 已插入到 5-HT1B R 的启动子和编码区 (tetO1B) 之间。原位杂交显示，与 129 只 WT 小鼠相比，这些 tetO1B 小鼠的大脑中具有正常的基础 5-HT1B R 表达。转基因小鼠在 ¿ 的控制下表达四环素依赖性转录沉默子 (tTS) -肌动蛋白（普遍存在）、CaMKII（前脑特异性）或 Pet（中缝特异性）启动子产生具有组织特异性和可诱导性敲低 5-HT1B R 的小鼠品系。放射自显影和原位杂交的初步表征揭示了启动子特异性 5 -HT1B R 表达模式可通过使用 tetO1B:: ¿ 的强力霉素进行调节。 -肌动蛋白-tTS+小鼠表现出与组成型基因敲除小鼠相似的行为表型，即冲动攻击性显着增加。因此，我们拥有在空间和时间上操纵 5-HT1B R 表达的工具，并检查由此产生的对攻击性的影响。使用光遗传学的后续研究将通过确定成年期的攻击性和冲动回路中的哪些大脑区域可能因关键时期的干预而改变来解决回路水平机制。在发育的关键时期缺乏 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，可以使用光敏抑制性 Cl-泵来挽救正常的表型，盐视紫红质，通过病毒载体注射到目标大脑区域，随后通过光纤植入物进行黄光激活。总体而言，了解 5-HT1B R 如何介导攻击性和冲动性的神经回路将具有重要意义。用于治疗一些神经精神疾病，包括成瘾、注意力缺陷多动障碍和双相情感障碍，以及自杀抑郁症。