Generating mouse models with cell type-specific and reversible GABA deficiency

生成具有细胞类型特异性和可逆性 GABA 缺陷的小鼠模型

• 批准号: 10679713
• 负责人: Xiaoxi Zhuang
• 金额: $ 23.73万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679713
• 关键词: Adult; Affect; Area; Basal Ganglia; Bypass; Communities; Corpus striatum structure; Cre driver; Disease; Disinhibition; Dopamine; Dopamine D1 Receptor; Dopamine D2 Receptor; Drosophila genus; Electrophysiology (science); Elements; Enzymes; Epilepsy; Gap Junctions; Genetic; Glutamate Decarboxylase; Glutamates; Goals; Health; Human; Injections; Internal Ribosome Entry Site; Interneurons; Knock-out; LoxP-flanked allele; Measures; Mediating; Memory; Modeling; Morphology; Motor; Motor Activity; Motor output; Mus; Mutation; National Institute of Mental Health; National Institute of Neurological Disorders and Stroke; Neonatal; Nervous System; Neurons; Neuropeptides; Neurosciences Research; Neurotransmitters; Parvalbumins; Pathway interactions; Physiological; Property; Pyridoxal Phosphate; Pyridoxine 5 Phosphate Oxidase; Research; Reversal Learning; Schizophrenia; Seizures; Short-Term Memory; Sleep; Synaptic Receptors; Testing; Tyrosine 3-Monooxygenase; Vitamin B6; brain morphology; cell type; cofactor; dietary; dopaminergic neuron; drinking; experimental study; feasibility testing; feeding; gamma-Aminobutyric Acid; inhibitory neuron; knockout gene; motor learning; mouse model; novel strategies; postsynaptic; presynaptic; response; tool; transmission process

ABSTRACT GABAergic neurons are the main inhibitory neurons in the adult nervous system. There are diverse cortical and subcortical GABAergic interneurons and projection neurons with different physiological properties, connectivity, and functions. However, few studies have been able to investigate the functions of specific subclasses by perturbing GABA release selectively. We propose an approach to target GABA synthesis in subclasses of GABAergic neurons. GAD activity requires the cofactor pyridoxal-5-phosphate (PLP). PLP deficiency can be achieved by knocking out the rate limiting enzyme in PLP synthesis: pyridoxine-5-phosphate oxidase (PNPO). Moreover, PLP deficiency can be rescued by systemic PLP administration. We therefore propose to generate cell type specific PNPO deficiency mouse models. We will first generate models with GABA deficiency in well-studied but highly distinct cell types to test the feasibility of our approach: 1) the parvalbumin (PV)-positive GABAergic neurons; 2) the VIP-positive GABAergic neurons; 3) the striatal dopamine D1 receptor expressing neurons; and 4) the striatal dopamine D2 receptor expressing neurons. Mice will be assessed for general health, feeding, drinking, sleep, and potential seizures. In addition, working memory, reference memory, reversal learning, motor coordination and motor learning will be examined. Brain morphology and electrophysiology will be assessed. Levels of GABA, other PNPO-dependent neurotransmitters, and their metabolites will be measured. The main goal of this R21 application is to test the feasibility of such an approach. These models are potentially significant research tools for the research community and may have a broad impact given the diverse distribution and functions of different GABAergic projection neurons and interneurons.

抽象的 GABA能神经元是成人神经系统中主要的抑制性神经元。皮质有多种 以及具有不同生理特性的皮层下 GABA 能中间神经元和投射神经元， 连接性和功能。然而，很少有研究能够调查特定的功能 通过选择性干扰 GABA 释放来形成子类。我们提出了一种靶向 GABA 合成的方法 GABA能神经元的亚类。 GAD 活性需要辅因子 5-磷酸吡哆醛 (PLP)。 PLP 可以通过敲除 PLP 合成中的限速酶：5-磷酸吡哆醇来解决缺乏症 氧化酶（PNPO）。此外，PLP 缺陷可以通过全身性 PLP 给药来挽救。我们因此 建议建立细胞类型特异性 PNPO 缺陷小鼠模型。我们将首先生成模型 经过充分研究但高度不同的细胞类型中 GABA 缺乏，以测试我们方法的可行性：1） 小清蛋白 (PV) 阳性 GABA 能神经元； 2）VIP阳性GABA能神经元； 3）纹状体 表达多巴胺 D1 受体的神经元； 4) 表达纹状体多巴胺 D2 受体的神经元。小鼠 将评估一般健康状况、进食、饮水、睡眠和潜在的癫痫发作。此外，工作 将检查记忆、参考记忆、逆向学习、运动协调和运动学习。脑 将评估形态学和电生理学。 GABA 水平，其他 PNPO 依赖性 神经递质及其代谢物将被测量。此 R21 应用程序的主要目标是测试 这种方法的可行性。这些模型可能是研究的重要研究工具 鉴于不同 GABAergic 的不同分布和功能，可能会产生广泛的影响 投射神经元和中间神经元。