Functional study of the role of SAPAP3 postsynaptic density protein on dorsolateral striatal cholinergic interneurons

SAPAP3突触后密度蛋白对背外侧纹状体胆碱能中间神经元作用的功能研究

• 批准号: 10606018
• 负责人: Alexander Theodor Baez
• 金额: $ 3.91万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-02 至 2026-09-01
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10606018
• 关键词: Acetylcholine; Acute; Age; Anxiety; Basal Ganglia; Behavior; Behavioral; Brain; Brain region; Cells; Cholinergic Receptors; Chronic; Compulsive Behavior; Corpus striatum structure; Data; Diagnosis; Disease; Dopamine; Dorsal; Electrophysiology (science); Excitatory Synapse; Face; Functional disorder; Gene Expression; Glutamate Receptor; Goals; Grooming; Home; Human; Immunohistochemistry; Interneuron function; Interneurons; Investigation; Knock-in; Knockout Mice; Laser Scanning Microscopy; Lead; Light; Link; Measures; Mediating; Membrane; Modeling; Motor; Movement; Mus; Mutation; N-Methyl-D-Aspartate Receptors; Neuromodulator; Neurons; Neurotransmitters; Obsessive-Compulsive Disorder; Output; Pathologic; Pathology; Pattern; Phase; Phenotype; Physiological; Physiology; Play; Population; Prevalence; Process; Property; Proteins; Role; Scaffolding Protein; Selective Serotonin Reuptake Inhibitor; Sensory; Shapes; Signal Transduction; Site; Slice; Source; Specificity; Substantia nigra structure; Synapses; Testing; Thalamic structure; Transferase; Trichotillomania; United States; Viral; Woman; base; behavioral phenotyping; cholinergic; density; early onset; genome wide association study; men; motor behavior; mouse model; neuroimaging; neuronal cell body; neuropsychiatric disorder; neuropsychiatry; neurotransmitter release; optogenetics; patch clamp; postnatal; postsynaptic; postsynaptic density protein; response; scaffold; sensor; targeted treatment; two photon microscopy; two-photon

PROJECT SUMMARY/ABSTRACT Obsessive Compulsive Disorder (OCD) is a neuropsychiatric disease listed in top 10 and top 15 most disabling illnesses in women and men, respectively, according to the Lancet in 2017 and has a predicted prevalence of 2.3% in the United States. OCD is characterized symptomatically in humans by obsessive thought patterns and compulsive motor behaviors. Synaptic deficits have been linked to psychiatric and neurodevelopmental diseases, including OCD, but the influence of disease-linked synaptic proteins on cellular, circuit, and behavioral outputs are incompletely understood. SAPAP3 is a synaptic protein, whose mutation is correlated with OCD diagnosis in humans. Constitutive SAPAP3 deletion in mice produces compulsive motor grooming behaviors, which are rescued by chronic administration of selective serotonin reuptake inhibitors (SSRIs) and striatum-localized SAPAP3 re-expression. The SAPAP3-deletion model of OCD, therefore, has symptomatic as well as treatment validity and can be used to investigate cellular- and circuit-level striatal dysfunctions underlying this compulsive motor phenotype. Preliminary evidence highlights striatal cholinergic interneurons (ChIs) as a likely contributor of widespread striatal dysregulation in this model. These cells display increased evoked release of Acetylcholine (ACh) in the SAPAP3-lacking striatum, which can modulate striatal circuits through myriad subtypes of widely expressed ACh receptors. This proposal combines electrophysiology, optogenetics, and 2-photon scanning laser microscopy (2PLSM) to test the overarching hypothesis, that functional dysregulation of ChIs is tied to maladaptive striatal ACh release and compulsive motor behavior in the SAPAP3-lacking model. In (Aim 1), patch-clamp electrophysiology in ex vivo brain slices will be used to compare intrinsic ChI function, and brain slice immunohistochemistry will be used to probe synaptic ACh release machinery. This will uncover SAPAP3 deletion’s impacts on intrinsic functional properties related to ACh release by ChIs. (Aim 2) will leverage viral optogenetic and ACh sensor constructs, electrophysiology, and 2PLSM to test for input-specific changes to synaptically evoked striatal ACh release. Finally, (Aim 3) will selectively rescue SAPAP3 expression in ChIs to test if this model’s disrupted striatal ACh release and OCD-like phenotype are intrinsically driven by SAPAP3-deletion in these cells. This will inform whether ChI-targeted therapies may be sufficient to modify this OCD-like circuit and behavioral phenotype.

项目摘要/摘要 强迫症（OCD）是一种神经精神疾病，在十大和最残疾的前15名中列出 根据2017年柳叶刀的说法，男女的疾病分别是 美国2.3％。强迫症通过强迫性思维模式在人类中的症状表征 和强迫运动行为。突触定义与精神病和神经发育有关 包括强迫症在内的疾病，但与疾病相关的合成蛋白对细胞，电路和 行为输出尚不完全理解。 SAPAP3是一种突触蛋白，其突变相关 与人类的强迫症诊断。小鼠的组成型SAPAP3删除会产生强迫性运动修饰 行为，通过长期给予选择性5-羟色胺再摄取抑制剂（SSRI）和 纹状体定位的SAPAP3重新表达。因此，强迫症的SAPAP3损坏模型具有症状 以及治疗有效性，可用于研究细胞和电路级纹状体功能障碍 这种强迫运动表型的基础。初步证据突出纹状体胆碱能中间神经元 （CHI）作为该模型中宽度纹状体失调的可能因素。这些细胞显示的增加 诱发乙酰胆碱（ACH）在SAPAP3占用纹状体中的释放，该纹状体可以调节纹状体电路 通过广泛表达的ACH受体的无数亚型。该建议结合了电生理学， 光遗传学和2光子扫描激光显微镜（2PLSM）以测试总体假设，即 CHI的功能失调与适应不良的纹状体ACH释放和强迫性运动行为有关 SAPAP3磁性模型。 在（AIM 1）中，将使用体内脑切片中的斑块夹电生理学来比较内在的CHI功能， 和脑切片免疫组织化学将用于探测突触释放机制。这将发现 SAPAP3删除对与CHIS释放有关的固有功能性能的影响。 （目标2）将 利用病毒光学遗传学和ACH传感器构建体，电生理学和2PLSM测试输入特异性 更改突触诱发的纹状体ACH释放。最后，（AIM 3）将有选择地拯救SAPAP3 在CHI中的表达以测试该模型的纹状体ACH释放和OCD样表型是否本质上是 由这些细胞中的SAPAP3删除驱动。这将告知卡靶疗法是否足以 修改此类似OCD的电路和行为表型。