Non-apoptotic functions of caspase-3 in neural development

Caspase-3在神经发育中的非凋亡功能

• 批准号: 10862033
• 负责人: Karina S Cramer
• 金额: $ 47.33万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10862033
• 关键词: Acceleration; Acoustic Nerve; Apoptosis; Apoptotic; Appearance; Auditory; Auditory Perception; Auditory system; Axon; Brain; Brain Stem; CASP3 gene; CASP6 gene; CASP8 gene; CASP9 gene; Caspase; Categories; Cell Death; Cell Death Signaling Process; Cell Membrane Permeability; Cell Nucleus; Cells; Cessation of life; Chick Embryo; Cochlea; Communication; Cytoprotection; Data; Defect; Dendrites; Development; Developmental Process; Electroporation; Embryo; Embryonic Development; Ganglia; Genes; Goals; Growth Associated Protein 43; Hearing Tests; Language; Mediating; Mitochondria; Modeling; Molecular; NCAM1 gene; Nerve; Neuron-Glia Cell Adhesion Molecules; Neurons; Neurosciences; Ontology; Outer Mitochondrial Membrane; Pathway interactions; Peptide Hydrolases; Permeability; Plasmids; Process; Proteins; Proteolysis; Proteome; Proteomics; Regulation; Research; Role; Sampling; Site; Source; Structural defect; Synapses; Systems Development; Testing; Time; Transfection; Transplantation; Work; auditory pathway; axon growth; axon guidance; cytotoxic; exosome; extracellular vesicles; neural circuit; neurodevelopment; receptor; sound; tandem mass spectrometry; vesicular release; x-linked inhibitor of apoptosis protein

PROJECT SUMMARY Auditory function relies on highly specialized and precise neuronal connectivity. A significant challenge for the field of auditory neuroscience is to understand how these neural circuits form during development. Our previous work suggests an important function for caspase-3, a protease best known for its role in apoptosis. Cleaved (active) caspase-3 is present in the developing auditory brainstem prior to the period of programmed cell death. During embryonic development, it is first seen in auditory nerve axons, then in the synaptic target of these axons in nucleus magnocellularis (NM), then in the synaptic target of NM, in nucleus laminaris (NL) dendrites. Caspase-3 inhibition during development results in substantial errors in NM axon targeting and in structural abnormalities in NL. We propose to investigate the regulation of caspase-3 activation during development. We will examine the basis for the progression of caspase-3 activation through the auditory pathway and test the hypothesis that cleaved caspase-3 is necessary in auditory axons for activation of caspase-3 in their synaptic targets. Caspase-3 activation during apoptosis is activated by cell death signals and mitochondrial permeabilization. We hypothesize that during auditory development, caspase-3 is activated through a non-canonical pathway that is protective for cells with cleaved caspase-3 in their axons. We will test the function of upstream molecules and determine whether they influence development of the NM-NL pathway. We have begun to investigate the molecules through which caspase-3 influences auditory development. Our proteomics study revealed hundreds of proteins that are cleaved by caspase-3 in the developing auditory brainstem. We have identified several substrates that mediate axon growth. We will use caspase-uncleavable forms of these proteins to test their caspase-dependent functions in development. Gene ontology analysis revealed that the most abundant cellular localization category for caspase-3 substrates was exosomes/extracellular vesicles (EVs). This finding suggests an overarching model in which caspase-3 influences the composition of EVs, which in turn provide an effective means of local communication between cells during development. We will examine enriched EV samples using tandem mass spectrometry to determine which caspase-3 substrates are present in EVs. We will use an EV grafting strategy to investigate whether EVs can rescue developmental deficits in caspase-3 inhibited host embryos. Together, these studies will advance our understanding of neural circuit assembly in the developing auditory brainstem.

项目摘要 听觉功能依赖于高度专业化和精确的神经元连接性。重要的 听觉神经科学领域的挑战是了解这些神经回路如何形成 发展。我们以前的工作暗示了caspase-3的重要功能，蛋白酶是最好的 以其在凋亡中的作用而闻名。开发听觉中存在裂解（活动）caspase-3 在编程细胞死亡期之前的脑干。在胚胎开发过程中，这是首先 在听觉神经轴突中可见，然后在这些轴突的突触靶标中看到。 （nm），然后在NM的突触靶中，在laminaris核（NL）树突中。 caspase-3抑制 在开发过程中会导致NM轴突靶向和结构异常的实质性误差 在NL。我们建议研究开发过程中caspase-3激活的调节。我们将 检查caspase-3激活通过听觉途径的基础并进行测试 在听觉轴突中需要裂解的caspase-3是激活caspase-3中必须的假设 他们的突触目标。细胞死亡信号激活凋亡过程中的caspase-3激活 线粒体通透性。我们假设在听觉开发过程中，caspase-3是 通过非典型途径激活，该途径可保护其在其中裂解caspase-3的细胞 轴突。我们将测试上游分子的功能，并确定它们是否影响 NM-NL途径的开发。我们已经开始研究分子 caspase-3影响听觉发展。我们的蛋白质组学研究揭示了数百种蛋白质 在发育的听觉脑干中被caspase-3裂解。我们已经确定了几个基板 介导轴突生长。我们将使用这些蛋白质的caspase毫不可见量的形式来测试 caspase依赖性功能在发育中。基因本体分析表明，最多 caspase-3底物的大量细胞定位类别是外泌体/细胞外囊泡 （EV）。这一发现表明了一个总体模型，其中caspase-3影响 电动汽车，这反过 发展。我们将使用串联质谱法检查富集的EV样品，以确定 EV中存在哪些caspase-3底物。我们将使用EV嫁接策略进行调查 电动汽车是否可以挽救caspase-3抑制宿主胚胎中的发育缺陷。在一起，这些 研究将提高我们对发展听觉中神经电路组件的理解 脑干。