Defining CAMK4 transcript isoforms for axonal plasticity

定义轴突可塑性的 CAMK4 转录亚型

• 批准号: 10317841
• 负责人: Jeffrey C Petruska
• 金额: $ 42.95万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10317841
• 关键词: 3' Untranslated Regions; Address; Adipocytes; Adult; Affect; Axon; Bioinformatics; Biomedical Research; Cells; Cerebellum; Characteristics; Code; Communities; Data; Development; Disease; Dysautonomias; Enterobacteria phage P1 Cre recombinase; Equilibrium; Foundations; Gene Expression Profile; Generations; Genes; Genetic Recombination; Genetic Transcription; Goals; Growth; Health Care Research; Hippocampus (Brain); In Situ Hybridization; Injury; Intrinsic factor; Kidney; Knock-out; Knockout Mice; Laboratories; LoxP-flanked allele; Mammals; Mediating; Molecular; Mus; Natural regeneration; Nervous System Trauma; Nervous system structure; Neurology; Neuronal Plasticity; Neurons; Neurosciences; Nociceptors; Northern Blotting; Pain; Pathology; Pattern; Process; Protein Isoforms; Proteins; Rattus; Recovery; Recovery of Function; Regulation; Research; Role; Sensory; Services; Stroke; T-Lymphocyte; Techniques; Testing; Testis; Tissues; Transcript; Untranslated Regions; axon growth; axon injury; axon regeneration; base; cell type; design; experimental study; mouse model; novel; podocyte; prevent; programs; protein expression; therapeutic target; tool; transcription factor; transcriptome sequencing

Affecting axonal growth as a means to enhance recovery and alleviate pathology in conditions of nervous system injury, insult, or disease is a major goal for the healthcare and biomedical research endeavors. Significant effort is directed at inducing neural plasticity to enhance axonal growth to establish functionally- adaptive connections. However, these efforts must also prevent, and not induce, maladaptive plasticity, a balance which requires a clear understanding of the processes regulating axon growth. We have recently determined that the mechanisms of the two major forms of axon growth in the adult nervous system – growth of injured axons (Regeneration) and that of non-injured axons (Collateral Sprouting - CS) – differ significantly and involve distinct transcriptional profiles. We have identified Camk4 as a gene necessary for Collateral Sprouting but not involved in Regeneration. We have further determined that during CS, Camk4 expression is regulated not at the coding sequence, but at the 3’ UTR. Camk4, and particularly Camk4 with this novel sprouting-related 3’ UTR, is expressed in neurons known to have a high degree of constitutive plasticity capacity such as sensory nociceptors, hippocampus, cerebellum, and cortex. These findings point to additional potential mechanisms by which CamK4 may be acting, and also opens additional potential therapeutic targets. However, tools to examine those mechanisms are lacking. We aim to determine the range and character of transcript isoforms expressed by neurons at basal state and during plasticity (regeneration and sprouting). These data will have significant value on their own, but will be used here to direct the design of a novel mouse with conditional-deletion of the novel long-isoform of the 3’ UTR without disrupting the protein coding sequence. This will allow us to determine the role of this important transcript segment in protein expression and localization, and examine other protein-independent functions which may exist. Although vital for mechanistic studies of axonal plasticity, these data and the new mice will also be useful for any cells which express Camk4 and use the novel long 3’ UTR (e.g., testicular cells, kidney podocytes, adipocytes, T cells, etc.) and we have established collaborative arrangements with a range of labs that would like to use these new mice in their research.

影响轴突生长作为促进恢复和缓解紧张状态下病理的一种手段 系统损伤、侮辱或疾病是医疗保健和生物医学研究工作的主要目标。 大量努力旨在诱导神经可塑性以增强轴突生长以建立功能性 然而，这些努力还必须防止而不是诱发适应不良的可塑性。 平衡，需要清楚地了解调节轴突生长的过程。 确定了成人神经系统中轴突生长的两种主要形式的机制——生长 受伤的轴突（再生）和未受伤的轴突（侧枝发芽 - CS） – 差异显着 并涉及不同的转录谱，我们已确定 Camk4 是抵押品所必需的基因。 我们进一步确定，在CS过程中，Camk4表达是发芽但不参与再生。 不是在编码序列上进行调节，而是在 3' UTR 上进行调节，尤其是在该小说中的 Camk4 上。 与发芽相关的 3' UTR 在已知具有高度组成可塑性的神经元中表达 这些发现指出了其他功能，例如感觉伤害感受器、海马体、小脑和皮质。 CamK4 可能发挥作用的潜在机制，并且还开辟了其他潜在的治疗靶点。 然而，我们缺乏检查这些机制的工具，以确定其范围和特征。 神经元在基础状态和可塑性（再生和萌芽）期间表达的转录亚型。 这些数据本身就具有重要价值，但将在这里用于指导新型鼠标的设计 有条件地删除 3' UTR 的新型长亚型，而不破坏蛋白质编码 这将使我们能够确定这个重要转录片段在蛋白质表达中的作用。 和定位，并检查可能存在的其他与蛋白质无关的功能，尽管对于这些功能至关重要。 轴突可塑性的机制研究，这些数据和新小鼠也将适用于任何细胞 表达 Camk4 并使用新型长 3’ UTR（例如睾丸细胞、肾足细胞、脂肪细胞、T 细胞、 等），我们已经与一系列希望使用这些新的实验室建立了合作安排 老鼠在他们的研究中。