Defining CAMK4 transcript isoforms for axonal plasticity

影响轴突生长作为促进神经性疾病恢复和减轻病理改变的手段 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。 大量的努力旨在诱导神经可塑性以促进轴突生长以建立功能性- 自适应连接。然而，这些努力也必须防止而不是诱导适应不良的可塑性， 平衡，这需要清楚地了解调节轴突生长的过程。我们最近做了 确定了成人神经系统中轴突生长的两种主要形式-生长的机制 损伤轴突(再生)和非损伤轴突(侧枝萌发-CS)的差异显著 并涉及不同的转录图谱。我们已经确定Camk4是一种侧枝必需的基因 萌芽的，但不参与再生的我们进一步确定在CS期间，Camk4的表达是 不是在编码序列上调节，而是在3‘非编码区调节。Camk4，特别是Camk4与这部小说 发芽相关的3‘非编码区，在已知具有高度结构可塑性的神经元中表达 感觉伤害性感受器、海马体、小脑和皮质。这些发现指出了其他 CamK4可能发挥作用的潜在机制，也开辟了更多潜在的治疗靶点。 然而，目前还缺乏研究这些机制的工具。我们的目标是确定范围和特征 神经元在基础状态和可塑性(再生和萌发)过程中表达的转录异构体。 这些数据本身将具有重要的价值，但将在这里用于指导新型鼠标的设计 在不破坏蛋白质编码的情况下有条件地删除新的3‘非编码区长异构体 序列。这将使我们能够确定这个重要的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蛋白质非依赖功能。虽然对 轴突可塑性的机制研究，这些数据和新的小鼠也将对任何 表达Camk4并使用新的长3‘非编码区(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技术的实验室建立了协作安排 老鼠在他们的研究中。

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.