Defining CAMK4 transcript isoforms for axonal plasticity

影响轴突生长作为一种手段，以加强恢复和减轻病理条件下的神经 系统损伤、损伤或疾病是保健和生物医学研究努力的主要目标。 显著的努力是针对诱导神经可塑性，以增强轴突生长，以建立功能性神经元。 适应性连接然而，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 确定了成年神经系统中轴突生长的两种主要形式的机制-生长 损伤的轴突（再生）和未损伤的轴突（侧支发芽- CS）-差异显着 并涉及不同的转录谱。我们已经确定Camk 4是一个必要的基因， 发芽但不参与再生。我们已经进一步确定，在CS期间，Camk 4表达是 不是在编码序列上，而是在3' UTR上调节。Camk 4，尤其是Camk 4， 发芽相关的3' UTR，在已知具有高度组成性可塑性的神经元中表达 例如感觉伤害感受器、海马、小脑和皮层。这些发现表明， 这些研究揭示了CamK 4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，缺乏审查这些机制的工具。我们的目标是确定的范围和性质 在基础状态和可塑性（再生和发芽）期间由神经元表达的转录物同种型。 这些数据本身将具有重要的价值，但在此将用于指导新颖鼠标的设计 在不破坏蛋白质编码的情况下， 顺序这将使我们能够确定这一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独立功能。虽然重要的是 轴突可塑性的机制研究，这些数据和新的小鼠也将有助于任何细胞， 表达Camk 4并使用新的长3’UTR（例如，睾丸细胞，肾足细胞，脂肪细胞，T细胞， 等等）。我们已经与一系列希望使用这些新技术的实验室建立了合作协议， 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.