Impact of Single-Walled Carbon Nanotubes on Growth and Reproductive Parameters in Fish

摘要#1236029 Sabo-Attwood，TaraNanomaterials（NM）已经成为高性能实体，在许多领域都有应用，包括电子，光学，医疗和结构复合材料技术。由于它们在大规模工业生产中的预期作用，毫无疑问，选择NM最终将进入我们的水生环境。目前还没有基础研究调查单壁碳纳米管（SWNT）生物相互作用的影响和对水生生物的影响，特别是通过慢性饮食接触途径，更少的是解决了作用机制。因此，我们有有限的了解与环境相关的exposs. This建议的目的是从根本上了解单壁碳纳米管的亚致死效应，重点是阐明他们的作用，限制生长和繁殖大口黑鲈（LMB），一个很好的特点与环境相关的淡水模型的生殖毒理学测定。我们特别感兴趣的是调查原始的非表面功能化的单壁碳纳米管的能力，改变胃肠道（GI）环境，通过直接组织损伤和/或螯合蛋白质的消化和吸收的饮食成分。研究设计涉及一种系统的自上而下的方法，涵盖了体内鱼类暴露于体外试验的机制。我们将通过以下具体任务完成这些研究：（1）使用定制的近红外荧光（NIRF）成像系统跟踪SWNT分布，并评估饮食暴露于LMB后的生长;（2）评估SWNT和富含雌激素的饮食对生殖终点的影响，包括卵黄蛋白原和类固醇的测量（17& #946;（3）采用蛋白质组学方法，测定肠组织和血液中与单壁碳纳米管结合的蛋白质组。这项拟议的研究将填补科学文献中的一个关键空白，提供对摄入单壁碳纳米管的生长和生殖影响的系统了解。这也将是第一个通过采用一套分子测定和蛋白质组学策略来解决特定作用机制（MOA）的研究。 这些研究将是完全新颖的，因为没有基本的机制报告已经检查了SWNT通过破坏鱼类的胃肠道和内分泌系统对生长和生殖过程的影响。这项研究的结果可能会导致在水生环境中的单壁碳纳米管的生物学关系，允许其命运和影响的先验预测，因此符合美国国家科学基金会提出的定义为变革性研究。预期的研究成果和益处包括在整个动物中系统跟踪单壁碳纳米管;深入了解慢性饮食暴露于单壁碳纳米管对内分泌系统的影响;揭示单壁碳纳米管与生物分子和特定MOA的相互作用，包括受体激活和类固醇的细胞产生。基础科学知识的增加将最终导致了解相关的环境接触情景和可能影响种群一级水生生物的亚致死生物效应。更广泛的影响。拟议的活动将产生关键知识，以更好地了解一类具有重要商业意义的纳米材料的环境影响。大部分申请的资金都是针对一个新兴的跨学科研究课题的博士后助理的培训。这项研究将提供指导和交叉培训（分子信号和蛋白质组学）在一个新的和高度相关的领域，是直接和至关重要的，我们的社会。博士后培训计划整合了课堂和动手实验室培训，并有机会指导被招募到实验室工作的少数民族研究生和本科生。计划通过会议介绍、同行评议出版物和研究专题讨论会传播研究成果。

Abstract#1236029Sabo-Attwood, TaraNanomaterials (NM) have emerged as high-performance entities with application in numerous sectors, including electronic, optical, medical, and structural composite technologies. Because of their anticipated role in large-scale industrial production, there is little doubt that select NM will ultimately find their way into our aquatic environment. There is a lack of fundamental studies that have investigated the effects of single-walled carbon nanotube (SWNT) biological interactions and effects in aquatic organisms, particularly through chronic dietary exposure routes, and even fewer that have addressed mechanisms of action. Therefore we have limited understanding of the potential toxicity associated with environmentally relevant exposures.The purpose of this proposal is to fundamentally understand the sub-lethal effects of SWNTs with a focus on elucidating their role in limiting growth and reproduction in largemouth bass (LMB), a well characterized environmentally relevant freshwater model for reproductive toxicological assays. We are particularly interested in investigating the ability of pristine non-surface functionalized SWNTs to alter the gastrointestinal (GI) environment through direct tissue damage and/or sequestering proteins vital to digestion and absorption of dietary constituents. The study design involves a systematic top down approach spanning in vivo fish exposures to mechanistic in vitro assays. We will accomplish these studies through the following specific tasks; (1) Track SWNT distribution using a custom built near-infrared fluorescent (NIRF) imaging system and assess growth following dietary exposures to LMB; (2) Assess the impact of SWNTs and estrogen enriched diets on reproductive endpoints including measurements of vitellogenin and steroids (17β-estradiol and 11-keto-testosterone) in plasma by ELISA and determination of estrogen receptor binding and activation using fluorescence polarization and gene reporter assays, respectively; (3) Determine suites of proteins bound to SWNTs in intestinal tissues and blood using proteomics strategies. The proposed research will fill a critical gap in the scientific literature by providing a systematic understanding of the growth and reproductive effects of ingested SWNTs. This will also be the first study to address specific mechanisms of action (MOA) by employing a suite of molecular assays and proteomic strategies. These studies will be fully novel, as there are no fundamental mechanistic reports that have examined the impact of SWNTs on growth and reproductive processes through disruption of the GI tract and endocrine system in fish. Results of this study may lead to biological relations for SWNTs in aquatic environments, allowing a priori predictions of their fate and effects and therefore meets the definition set forth by the NSF for transformative research.Intellectual Merit. The expected research outcomes and benefits include systemic tracking of SWNTs in whole animals; advanced understanding of chronic dietary exposures to SWNT on the endocrine system; revealing the interactions of SWNTs with biological molecules and specific MOA including receptor activation and cell production of steroids . This increase in basic scientific knowledge will ultimately lead to understanding relevant environmental exposure scenarios and sub-lethal biologic effects that could impact aquatic organisms at the population level. Broader Impacts. The proposed activity will generate critical knowledge to better understand the environmental implication of a commercially important class of nanomaterials. The majority of requested funds are directed toward the training of a postdoctoral associate in an emerging and interdisciplinary research topic. This research will provides for mentoring and cross training (molecular signaling and proteomics) in a novel and highly relevant area that is of immediate and critical importance to our society. The postdoctoral training program integrates classroom and hands on laboratory training and the opportunity to mentor minority graduate and undergraduate students recruited to work in the lab. Dissemination of the research results is planned through conference presentations, peer-reviewed publications and research symposiums.