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.

摘要 GABA能神经元是成人神经系统的主要抑制性神经元。有不同的皮质 皮质下GABA能中间神经元和投射神经元具有不同的生理特性， 连接性和功能。然而，很少有研究能够调查特定的功能 通过干扰GABA选择性释放的亚类。我们提出了一种靶向GABA合成的方法 GABA能神经元的亚类。GAD活性需要辅因子吡哆醛-5-磷酸(PLP)。PLP 缺乏可以通过敲除PLP合成中的限速酶：吡哆醇-5-磷酸来实现 氧化酶(PNPO)。此外，PLP缺乏症可以通过全身应用PLP来挽救。因此，我们 建议建立细胞类型特异性PNPO缺乏症小鼠模型。我们将首先使用以下内容生成模型 在研究充分但高度不同的细胞类型中存在GABA缺陷，以测试我们方法的可行性：1) Pv阳性的GABA能神经元；2)VIP阳性的GABA能神经元；3)纹状体 多巴胺D2受体阳性神经元；纹状体多巴胺D2受体阳性神经元。老鼠 将对一般健康、进食、饮水、睡眠和潜在的癫痫发作进行评估。此外，工作 将检查记忆、参考记忆、反转学习、运动协调和运动学习。脑区 将对形态学和电生理学进行评估。GABA和其他PNPO依赖的水平 神经递质及其代谢物将被测量。这个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 社区和可能产生广泛的影响，因为不同的GABA能的不同的分布和功能 投射神经元和中间神经元。