Proteomic analysis of maturing adult-born hippocampal mossy fiber boutons

成年海马区新神经元的诞生(称为神经发生)对学习和记忆至关重要 这一过程的中断与阿尔茨海默病等人类神经疾病有关。费率 成人海马区神经发生与生理活动的变化密切相关。活动，如 随着运动或学习的加强以及癫痫等病理生理变化，安氏将深刻改变。 因此，了解ANH神经发生是如何调节神经元回路的，对于全面了解其 对大脑生理的影响。这个问题的核心是了解成年大脑中新生的神经元如何实现 长期融合。了解AHN中调节突触整合的分子和细胞机制 可能导致选择性的药理靶点，在病理条件下或在 成体神经发生水平显著降低的衰老。成人的神经前体细胞 海马齿状回产生新生颗粒(GCs)细胞，当完全分化时，接收 来自内嗅皮层的突触输入，并沿着苔藓纤维通路发送轴突形成突触输出 有CA3锥体神经元。我们和其他人已经证明，新生儿GC大约需要8周的时间 以充分区分和形成成熟的突触输入和输出。这项提案的主要重点是确定 GC新生苔藓纤维环形成突触时突触输出的分子变化 有成熟的CA3锥体细胞。我们建议使用超分辨率免疫荧光阵列断层扫描和 共轭阵列断层扫描结合电子显微镜研究前、后细胞的蛋白质组变化 成熟突触建立过程中的突触后成分。我们发现，要建立一个成熟的 苔藓纤维可以1)形成新生突触，或者2)取代已有的苔藓 纤维连接蛋白控制已有的突触后CA3树突。调控的分子机制 这些不同的细胞过程是未知的。在这里，我们将使用阵列断层扫描分析来分析 这些突触的蛋白质组学变化，以检验突触分子组成 在整个成熟过程中，成年海马区新生神经元的整合是高度动态的。我们 主要关注两个方面：(1)建立发育和成熟的突触前苔藓纤维的蛋白质组学图谱 在成年海马神经发生过程中的终末和(2)建立发育和 成年海马神经发生过程中成熟的突触后苔藓纤维终末。这些实验将是 第一个解决在成人期间建立新的突触输出所必需的复杂的蛋白质组变化 神经发生，并可能确定一个治疗策略的药理靶点，以改善 成人脑的功能。

The birth of new neurons (called neurogenesis) in the adult hippocampus is critical for learning and memory and disruption of this process is associated with human neurological disorders such as Alzheimer’s disease. Rates of adult hippocampal neurogenesis (AHN) are tightly linked with changes in physiological activity. Activities such as enhanced exercise or learning as well as pathophysiological changes such as epilepsy, profoundly alter AHN. Knowing how ANH neurogenesis regulates neuronal circuitry is therefore important for understanding its overall impact on brain physiology. Central to this issue is understanding how newborn neurons in adult brain achieve long-term integration. Understand the molecular and cellular mechanisms regulating synaptic integration in AHN could lead to selective pharmacological targets for functional improvement during pathological conditions or in aging where levels of adult neurogenesis are dramatically decreased. Neuronal progenitors in the adult hippocampal dentate gyrus give rise to newborn granule (GCs) cells that, when fully differentiated, receive synaptic inputs from entorhinal cortex and send axons along the mossy fiber pathway to form synaptic outputs with CA3 pyramidal neurons. We and others have shown that it takes about eight weeks for the newborn GCs to fully differentiate and form mature synaptic inputs and outputs. The major focus of this proposal is to determine the molecular changes in the synaptic outputs when newborn mossy fiber boutons from GC are forming synapses with mature CA3 pyramidal cells. We propose to use superresolution immunofluorescent array tomography and conjugate array tomography coupled with electron microscopy to profile the proteomic changes of the pre- and post-synaptic elements during the establishment of mature synapses. We have found that to establish a mature synaptic contact the mossy fiber can either 1) form a de novo nascent synapse or 2) replace an existing mossy fiber bouton assuming control of the existing postsynaptic CA3 dendrite. The molecular mechanisms regulating these disparate cellular processes are unknown. Here we will use array tomography analysis to profile the proteomic changes in these synapses to test the hypothesis that the synaptic molecular composition of integrating newborn neurons in adult hippocampus is highly dynamic during the entire maturation process. We have two main focuses: (1) to establish a proteomic profile of the developing and mature presynaptic mossy fiber terminal during adult hippocampal neurogenesis and (2) to establish a proteomic profile of the developing and mature postsynaptic mossy fiber terminal during adult hippocampal neurogenesis. These experiments will be the first to address the intricate proteomic changes essential for establishing new synaptic outputs during adult neurogenesis and will potentially identify a pharmacological target for therapeutic strategies to improve the function of adult brain.