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中主要的抑制性神经元。大脑皮层有不同的 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， 通过干扰GABA的选择性释放的子类。我们提出了一种靶向GABA合成的方法， GABA能神经元的亚类。GAD活性需要辅因子吡哆醛-5-磷酸（PLP）。PLP 可以通过敲除PLP合成中的限速酶：5-磷酸吡哆素来实现缺乏 氧化酶（PNPO）。此外，PLP缺乏症可以通过全身性PLP给药来挽救。因此我们 提出产生细胞类型特异性PNPO缺陷小鼠模型。我们将首先生成模型， GABA缺乏症在充分研究，但高度不同的细胞类型，以测试我们的方法的可行性：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 多巴胺D1受体表达神经元;和4）纹状体多巴胺D2受体表达神经元。小鼠 将评估一般健康状况、进食、饮水、睡眠和潜在癫痫发作。此外，工作 将检查记忆、参考记忆、反向学习、运动协调和运动学习。大脑 将评估形态学和电生理学。GABA水平，其他PNPO依赖性 将测量神经递质及其代谢物。此R21应用程序的主要目标是测试 这种做法的可行性。这些模型是潜在的重要的研究工具的研究 社区，并可能具有广泛的影响，因为不同的GABA能的分布和功能不同， 投射神经元和中间神经元。