GENERATION OF MICE WITH CREB-DEFICIENT NEURONS

描述（改编自申请人摘要）：在神经系统中， 兴奋性神经传递显著影响基因的表达 可能对长期神经适应性反应至关重要， 包括LTP和LTD的维护阶段。因此， 神经递质敏感基因可能是复合物的重要底物 学习和记忆的神经生物学现象。 转录 CREB因子参与调节神经递质敏感性 基因和长期神经元适应性反应的控制 无脊椎动物和脊椎动物。 兴奋性神经递质刺激 CREB在单个转录调控位点丝氨酸磷酸化 133（Ser133），这种磷酸化事件对于CREB介导的 转录。 然而，可用于确定哺乳动物的免疫应答的哺乳动物模型系统是不理想的。 CREB在神经递质信号传导中的作用，或用于识别基因 其表达式是CREB依赖的。 一只老鼠， CREB基因内存在靶向突变，但这只小鼠表达高 脑和其他组织中功能性CREB蛋白的水平，以及 来自这些小鼠的神经元显示神经递质敏感的CREB介导的 转录。 本提案的总体目标是， 两个转基因小鼠系，其在CREB DNA结合中缺陷， 活性或表达突变CREB蛋白的细胞，所述突变CREB蛋白不能被 磷酸化 由于CREB依赖的转录可能是 神经系统发育的重要方面，我们将采用新的 表达我们先前描述的显性负性CREB的程序 转基因在有条件的情况下只在成年神经元中表达。 这些 老鼠将成为未来实验的基础， CREB Ser133磷酸化在神经递质调节中的作用 基因表达，突触可塑性，最终， 神经生物学现象，如学习和记忆。 此外，老鼠 仅在成年神经元中缺乏CREB活性将提供一种模型， 用于鉴定其表达依赖于 CREB。

DESCRIPTION (Adapted from applicant's abstract): In the nervous system, excitatory neurotransmission markedly influences the expression of genes that are likely to be critical for long-term neuronal adaptive responses, including the maintenance phases of LTP and LTD. Thus, neurotransmitter-sensitive genes may be important substrates of complex neurobiological phenomena underlying learning and memory. The transcription factor CREB has been implicated in regulation of neurotransmitter-sensitive genes and the control of long-term neuronal adaptive responses in both invertebrates and vertebrates. Excitatory neurotransmitters stimulate the phosphorylation of CREB on a single transcriptional regulatory site, serine 133 (Ser133), and this phosphorylation event is critical for CREB-mediated transcription. However, a mammalian model system useful for determining the role of CREB in neurotransmitter signaling, or for identification of genes whose expression is CREB-dependent does not exist. A mouse containing a targeted mutation within the CREB gene exists, but this mouse expresses high levels of a functional CREB protein in brain and other tissues, and the neurons from these mice display neurotransmitter-sensitive, CREB-mediated transcription. The overall objective of the present proposal is to generate two transgenic mouse lines that are either deficient in CREB DNA binding activity or that express a mutant CREB protein that cannot be activated by phosphorylation. Since CREB-dependent transcription is likely to be important for aspects of nervous system development, we will employ new procedures to express our previously characterized dominant negative CREB transgenes exclusively in adult neurons under conditional fashion. These mice will form the basis for future experiments that will address the role of CREB and phosphorylation of CREB Ser133 in neurotransmitter regulation of gene expression, synaptic plasticity and, ultimately, complex neurobiological phenomena such as learning and memory. Moreover, mice deficient in CREB activity exclusively in adult neurons will provide a model system useful for identification of gene whose expression is dependent upon CREB.