O-GlcNac Modulation of GABAergic Transmission

O-GlcNac 对 GABA 能传输的调节

• 批准号: 10754746
• 负责人: Shekinah Phillips
• 金额: $ 4万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10754746
• 关键词: Acute; Address; Affect; Allopregnanolone; Alzheimer' s Disease; Anti-Anxiety Agents; Anxiety; Award; Barbiturates; Behavior; Benzodiazepines; Binding; Biological Assay; Biology; Bipolar Disorder; Brain; Cells; Chloride Channels; Chronic; Communication; Cyclic AMP-Dependent Protein Kinases; Data; Depressed mood; Diabetes Mellitus; Diabetic mouse; Disease; Disease model; Distant; Electrophysiology (science); Endocytosis; Endocytosis Induction; Environment; Epigenetic Process; Epilepsy; Equilibrium; Exhibits; Frequencies; Functional disorder; GABA Modulators; Gene Expression; Genes; Genetic Polymorphism; Genome; Hippocampus; Histone Code; Impairment; Individual; Lead; Ligands; Link; Location; Long-Term Depression; Major Depressive Disorder; Mass Spectrum Analysis; Mediating; Mental Depression; Metabolic Diseases; Modeling; Modification; Mutation; Neurodegenerative Disorders; Neurons; Phase; Phosphorylation; Post-Translational Protein Processing; Postdoctoral Fellow; Postpartum Depression; Proteins; Publishing; Pyramidal Cells; Regulation; Reporting; Research; Research Project Grants; Role; Schizophrenia; Serine; Shapes; Site; Slice; Steroids; Synapses; Syndrome; Techniques; Testing; Therapeutic; Threonine; Training; Transfection; Transferase; Western Blotting; Work; X-linked intellectual disability; alcohol use disorder; clinically relevant; db/db mouse; diabetic; drug of abuse; epigenetic regulation; epigenome; gamma-Aminobutyric Acid; granule cell; hypnotic; in vivo; induced pluripotent stem cell; insight; mouse model; nervous system disorder; neurodevelopment; neuropsychiatric disorder; neurosteroids; novel; patch clamp; peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase; pharmacologic; positive allosteric modulator; receptor; receptor function; restoration; sedative; synaptic function; synaptic inhibition; therapeutic development; transcriptome; transmission process; Acute; Address; Affect; Allopregnanolone; Alzheimer' s Disease; Anti-Anxiety Agents; Anxiety; Award; Barbiturates; Behavior; Benzodiazepines; Binding; Biological Assay; Biology; Bipolar Disorder; Brain; Cells; Chloride Channels; Chronic; Communication; Cyclic AMP-Dependent Protein Kinases; Data; Depressed mood; Diabetes Mellitus; Diabetic mouse; Disease; Disease model; Distant; Electrophysiology (science); Endocytosis; Endocytosis Induction; Environment; Epigenetic Process; Epilepsy; Equilibrium; Exhibits; Frequencies; Functional disorder; GABA Modulators; Gene Expression; Genes; Genetic Polymorphism; Genome; Hippocampus; Histone Code; Impairment; Individual; Lead; Ligands; Link; Location; Long-Term Depression; Major Depressive Disorder; Mass Spectrum Analysis; Mediating; Mental Depression; Metabolic Diseases; Modeling; Modification; Mutation; Neurodegenerative Disorders; Neurons; Phase; Phosphorylation; Post-Translational Protein Processing; Postdoctoral Fellow; Postpartum Depression; Proteins; Publishing; Pyramidal Cells; Regulation; Reporting; Research; Research Project Grants; Role; Schizophrenia; Serine; Shapes; Site; Slice; Steroids; Synapses; Syndrome; Techniques; Testing; Therapeutic; Threonine; Training; Transfection; Transferase; Western Blotting; Work; X-linked intellectual disability; alcohol use disorder; clinically relevant; db/db mouse; diabetic; drug of abuse; epigenetic regulation; epigenome; gamma-Aminobutyric Acid; granule cell; hypnotic; in vivo; induced pluripotent stem cell; insight; mouse model; nervous system disorder; neurodevelopment; neuropsychiatric disorder; neurosteroids; novel; patch clamp; peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase; pharmacologic; positive allosteric modulator; receptor; receptor function; restoration; sedative; synaptic function; synaptic inhibition; therapeutic development; transcriptome; transmission process

Changes in the strength of GABAergic transmission is heavily influenced by posttranslational modifications and allosteric modulators like benzodiazepines and neurosteroids. O-GlcNAcylation (O- GlcNAc) is a post- translational modification that is tightly regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), which add or remove the O-GlcNAc moiety of β–N-acetylglucosamine to Ser/Thr residues on proteins, respectively. Various neurodegenerative diseases like Alzheimer's disease (AD), and metabolic disorders like, diabetes exhibit dysregulated O-GlcNAc levels. Published reports from our lab have demonstrated that an acute increase in O-GlcNAcylation induces a long-term depression of evoked GABAAR mediated IPSCs (eIPSCs) and reduces the amplitude and frequency of spontaneous IPSC (sIPSC) in hippocampal principal cells, however the mechanism in which this occurs is unknown. Numerous studies have shown that serine phosphorylation of GABAAR can increase or decrease GABAAR currents depending on the neuron type and specific subunit. While O-GlcNAcylation modulates inhibitory GABA-gated currents, no studies have examined its interplay with serine phosphorylation on GABAergic transmission. Because the crosstalk between O-GlcNAcylation and phosphorylation affects the regulation of various proteins, the potential exists that O-OglcNAcylation and phosphorylation will interaction in the modulation of GABAAR function and the strength of inhibitory transmission. Furthermore, a potential interaction could impact how allosteric modulators effect GABAARs, since serine phosphorylation can either increase or decrease efficacy of these modulators. Studies proposed will investigate these hypotheses and will also test whether disease conditions where O- GlcNAc levels are chronically elevated lead to depressed GABAAR function that can be rescued via pharmacological inhibition of OGT. To determine O-GlcNAcylation's effect on GABAergic transmission in the presence of phosphorylation and allosteric modulators, I will use electrophysiological techniques, immunoblotting assays and mass spectrometry. Results will show whether the modulation of GABAAR function by PKA and allosteric modulators are shaped by the presence or absence of a co-occurring O- GlcNAc modification and if the restoration of O-GlcNAcylation levels in a disease models can restore the E/I balance.

GABA能传播强度的变化受翻译后修饰和变构调节剂（如苯二氮卓类药物和神经类固醇）的严重影响。 O-GlcNAcylation (O-GlcNAc) is a post-translational modification that is tightly regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), which add or remove the O-GlcNAc moiety of β-N-acetylglucosamine to Ser/Thrresidues on proteins, Various neurodegenerative diseases like Alzheimer's disease （AD）和代谢性疾病，例如糖尿病暴露失调的O-GLCNAC水平。我们实验室的发表报告表明，O-Glcnacylation的急性增加会影响唤起的Gabaar介导的IPSC（EIPSCS）的长期抑郁症，并减少了发言人IPSC（SIPSC）（SIPSC）（SIPSC）的放大器和频率，但是在这一机制中，这是事实的。大量研究表明，Gabaar的丝氨酸磷酸化可以根据神经元类型和特定亚基增加或减少Gabaar电流。尽管O-Glcnacylation调节了抑制性GABA门控电流，但尚无研究检查其与GABA能传播中丝氨酸磷酸化的相互作用。由于O-Glcnacylation和磷酸化之间的串扰会影响各种蛋白质的调节，因此存在O-OGLCNACYLATION和磷酸化的潜力将在调节Gabaar功能和抑制性透射强度时相互作用。此外，由于丝氨酸磷酸化可以提高或降低这些调节剂的有效性，因此潜在的相互作用可能会影响变构调节剂的影响Gabaar。提出的研究将研究这些假设，还将测试O-GLCNAC水平长期升高的疾病状况是否会导致Gabaar功能降低，这可以通过OGT的药物抑制来挽救。为了确定O-Glcnacylation在磷酸化和变构调节剂存在下对GABA能传播的影响，我将使用电生理技术，免疫印迹测定和质谱法。结果将显示PKA对GABAAR功能的调节和变构调节剂的调节是否是由于存在或不存在共发生的O-GLCNAC修饰而塑造的，以及疾病模型中O-Glcnacylation水平的恢复是否可以恢复E/I平衡。