Investigating the function of localized BMP signaling at neuromuscular synapses

项目摘要/摘要 阐明神经元突触和膜电压变化的分子机制 将这些变化与适当的细胞反应联系起来，对生理学来说至关重要，因为神经电路必须 在适应新条件的同时保持动态平衡，以及与神经学和 神经退行性疾病，其中许多最终是由突触传递缺陷引起的。果蝇 黑素胃素已成为剖析这类机制的强大遗传模型。虽然意义重大 在识别突触的结构成分以及所使用的分子方面取得了进展 通过突触前和突触后细胞通过突触裂隙进行交流，这些细胞 细胞过程与膜电压的瞬时变化结合在一起仍然知之甚少。这个 目前的提案将通过关注一种非规范的局部化形式的骨如何解决这一知识缺口 突触上的形态发生蛋白(BMP)信号是膜电压的传感器，而 这种形式的信号有可能在瞬时膜电压变化和 更稳定的细胞变化。我们将测试三个中心假设：1)局部BMP信号响应 突触的电压变化；2)突触BMP信号的动态升降 膜电压的变化；以及3)突触BMP信号复合体与一套不同的 调节性和效应性分子。这些假说将通过使用体内遗传学、 基因编辑、免疫荧光显微镜、电子显微镜、分子生物学、生物化学和 电生理学方法。这些研究的短期影响将是对分子的新见解 果蝇细胞膜电压变化与细胞和分子变化联系的机制 神经肌肉接头突触。最终，这些研究可能确定进化保守的调控。 在人类突触生理学中具有重要作用的机制，并可能有助于我们的 了解这些细胞过程中的干扰如何构成神经和神经的病理生理学基础 神经退行性疾病状态。

Project Summary / Abstract Elucidating the molecular mechanisms that respond to membrane voltage changes at neuronal synapses and link these changes to the appropriate cellular responses is critical to physiology, as neural circuitry must maintain homeostasis while adapting to novel conditions, as well as in relation to neurological and neurodegenerative diseases, many of which ultimately result from defective synaptic transmission. Drosophila melanogaster has emerged as a powerful genetic model for dissecting such mechanisms. While significant progress has been made in identifying the structural components of synapses, as well as the molecules used by pre- and postsynaptic cells to communicate across the synaptic cleft, the mechanism by which these cellular processes integrate with transient changes in membrane voltage remain poorly understood. The current proposal will address this knowledge gap by focusing on how a noncanonical localized form bone morphogenetic protein (BMP) signaling at synapses serves as a sensor for membrane voltage, and the potential for this form of signaling to serve as an adapter between transient membrane voltage changes and more stable cellular alterations. We will test three central hypotheses: 1) that local BMP signaling responds to voltage changes at synapses; 2) that synaptic BMP signaling dynamically rises and falls in response to acute changes in membrane voltage; and 3) that synaptic BMP signaling complexes interact with a distinct suite of regulatory and effector molecules. These hypotheses will be tested by using a combination of in vivo genetics, gene editing, immunofluorescent microscopy, electron microscopy, molecular biology, biochemistry, and electrophysiology methods. The short-term impact of these studies will be novel insights into the molecular machinery linking membrane voltage changes with cellular and molecular alterations at Drosophila neuromuscular junction synapses. Ultimately, these studies may identify evolutionary conserved regulatory mechanisms with an important role in synaptic physiology in humans, and potentially contribute to our understanding of how disruptions in these cellular processes underlie the pathophysiology of neurological and neurodegenerative disease states.