Development of a behavioral rat model to assess proteomic and metabolomic adaptations of laryngeal muscles in response to vocal exercise

项目总结/摘要 随着大规模生物化学蛋白质组学和代谢组学方法的发展， 对神经肌肉可塑性反应中涉及的途径和生物标志物的深入理解 肢体肌肉组织的使用、废弃和老化。然而，喉肌蛋白质组和代谢组， 相对来说还未被探索过。如果不直接研究喉肌，诊断和治疗 治疗由推定的喉肌功能障碍引起的沟通障碍的方法仍然存在 理论上的推测。我们的长期目标是了解喉神经肌肉反应， 增加声音使用，如声音训练和声音治疗，减少声音使用，如声音休息 和衰老。这项建议的总体目标是确定蛋白质组和代谢组如何 甲杓肌对发声训练和年龄的反应。我们的中心假设是 发声训练将增加线粒体功能和氧化应激反应的信号通路 并且，相对于年龄，衰老将在甲杓肌中产生独特的蛋白质组学和代谢组学效应。 四肢肌肉这项工作将通过两个目标来完成。在雄性和雌性大鼠中，我们将描述 并确定甲杓肌中的特异性生物标志物和生化途径， 在以下两种情况下的神经肌肉适应：（1）发声训练和（2）衰老 （老化）。具体来说，我们将广泛地描述肌肉收缩特性，生物能量学和氧化还原应激 甲杓肌的肌肉组织的蛋白质和代谢物内的反应。这项工作将 建立在先前建立的行为动物模型上，包括训练大鼠以增加它们的 超声波发声此外，使用相同的蛋白质组学和代谢组学方法，我们将 通过检查甲状腺杓状肌，阐明年龄相关的变化如何在整个生命周期中表现出来。 年轻的成年人，老年人和衰老的雄性和雌性大鼠，并比较该肌肉中的生物标志物， 腓肠肌后肢中的生物标志物。该研究创新地使用了行为动物模型， 研究喉肌使用和衰老对神经肌肉机制的影响， 实施新型高通量蛋白质组学和代谢组学方法来检查喉 神经肌肉结构和功能。这些多组学方法将补充以前的研究 在我们的实验室中进行了关于正常和衰老啮齿动物模型的神经肌肉接头和肌纤维可塑性的研究， 从而为理解参与的细胞和分子神经肌肉通路奠定了基础。 正常和老化的喉内肌结构和功能。这项翻译工作将提供证据 用于临床人群的未来治疗目标，如老年性幻听和功能亢进的声音障碍。 这项工作与NIDCD早期职业研究奖的使命是一致的，因为我们建议 高度转化的研究，同时开发新的研究方法和技术。

Project Summary/Abstract The development of large-scale, biochemical proteomics and metabolomics approaches has led to a more sophisticated understanding of the pathways and biomarkers involved in neuromuscular plasticity in response to use, disuse, and aging in the limb musculature. The laryngeal muscle proteome and metabolome, however, remain relatively unexplored. Without direct study of the laryngeal muscles, diagnostic and therapeutic approaches for communication disorders caused by presumed laryngeal muscle dysfunction remain theoretically speculative. Our long-term goal is to understand the laryngeal neuromuscular response to increased voice use, such as vocal training and voice therapy, and decreased voice use, such as voice rest and senescence. The overall objective of this proposal is to determine how the proteome and metabolome of the thyroarytenoid muscle respond to vocalization training and aging. Our central hypothesis is that vocalization training will increase signaling pathways for mitochondrial function and oxidative stress responses and that aging will have unique proteomic and metabolomic effects in the thyroarytenoid muscle relative to the limb muscles. This work will be accomplished through two aims. In male and female rats we will characterize and identify specific biomarkers and biochemical pathways in the thyroarytenoid muscle indicative of neuromuscular adaptations under the following two conditions: (1) vocalization training and (2) senescence (aging). Specifically, we will broadly characterize muscle contractile properties, bioenergetics, and redox stress responses within the proteins and metabolites of muscle tissue of the thyroarytenoid muscle. This work will build on a previously established behavioral animal model involving training rats to increase their production of ultrasonic vocalizations. Additionally, using the same proteomic and metabolomic approaches, we will elucidate how age-related changes manifest across the lifespan by examining the thyroarytenoid muscle of young adult, older adult, and senescent male and female rats, and comparing biomarkers in this muscle to biomarkers in the gastrocnemius hindlimb. The study is innovative in its use of a behavioral animal model to investigate the effects of laryngeal muscle use and senescence on neuromuscular mechanisms and in its implementation of novel high-throughput proteomic and metabolomic approaches to examine laryngeal neuromuscular structure and function. These multi-omics approaches will complement previous studies conducted in our lab on neuromuscular junction and muscle fiber plasticity in normal and aging rodent models, thereby laying the foundation for understanding the cellular and molecular neuromuscular pathways involved in normal and aging intrinsic laryngeal muscle structure and function. This translational work will provide evidence for future therapeutic targets for clinical populations such as presbyphonia and hyperfunctional voice disorders. This work is consistent with the NIDCD’s mission for the Early Career Research Award in that we propose highly translational research while developing both novel research methodology and technology.