Nanoparticle Transport to Oocytes and Toxicological Consequences in Fathead Minnows

纳米颗粒向黑头呆鱼卵母细胞的运输及其毒理学后果

• 批准号: 0853707
• 负责人: Nancy Denslow
• 金额: $ 34万
• 依托单位: University of Florida
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2013-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0853707&HistoricalAwards=false
• 关键词: Nanoparticle; Transport; Oocytes; Toxicological; Consequences

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).0853707DenslowGrowing evidence suggests that many nanomaterials are taken up into fish species through the gills or digestive track. Once internalized, there is a potential for nanomaterials to cross into the blood stream, where they could pose a risk to internal organs including the ovary. Emerging literature and preliminary evidence in the current application suggest that nanomaterials may be capable of entering oocytes, potentially impacting embryonic development. There is a need to understand fundamental mechanisms that enable nanomaterial transport to oocytes, and the subsequent alterations of natural biological processes. It is currently understood that the physiological behavior of nanomaterials is largely impacted by adsorbed proteinaceous layers, which form upon seconds of exposure to biological media. Female fish that are actively reproducing have high serum levels (up to 20 mg/ml) of the egg yolk precursor protein, vitellogenin. This protein is a large phospholipoglycoprotein that has the capacity to bind to nanomaterials through various conduits, including areas of high negative charge and areas of high hydrophobicity. Vitellogenin is naturally targeted to developing oocytes and is imported into these oocytes via a vitellogenin-specific receptor. The overall hypothesis of this project is that vitellogenin-bound nanomaterials will be imported into yolk granules in developing oocytes and eventually merge with lysosomes. Furthermore, upon reaching this compartment certain nanomaterials could dissolve and leach into the cell causing toxicity for developing embryos. To address these hypotheses, the researchers have 3 specific aims: (1) Determine the effect of surface modification on uptake, cellular distribution and retention of nanomaterials in fish ovarian cultures. (2) Characterize the protein surface coating of nanomaterials exposed to male and female fathead minnow plasma. (3) Determine the distribution and reproductive toxicity of nanoparticles in vivo. To achieve these aims they will use model luminescent nanoparticulates, namely dye doped silica and quantum dots of carefully chosen size distributions and surface properties that will enable detailed analysis of the movement of these materials in vitro and in vivo. The proposed research will provide critical insights into the role of nanoparticulate properties on their translocation to oocytes. This information is anticipated to be highly important in the development of predictive models for nanotoxicity in fish species, and methods to mitigate potentially adverse outcomes of nanomaterial exposure to the environment. Cross-disciplinary training of graduate, undergraduate, and high school students at the interface between particle science and engineering and biology will be an essential part of the research program. The Southeast Alliance for Graduate Education and the Professoriate (SEAGEP), related Research Experience for Undergraduates (REU) programs, and established minority fellowship programs at the University of Florida will be leveraged to ensure diversity in the research program. Research results will be published in peer-reviewed journals and disseminated at national and international meetings. The investigators also plan to collaborate with national nanotechnology resources such as the Center for Nanotechnology in Society at Arizona State University and ICON at Rice to disseminate research results to the scientific and educational communities, and to society at large. The students will also participate in public outreach programs (e.g., UF's Engineering Fair)providing a conduit for them to engage the general public about the excitement of nanoscience and the importance of environmental stewardship. UF's Graham Center for Public Policy will be engaged to ensure that research findings are appropriately communicated to policy makers. The proposed research will provide fundamental knowledge of factors that mediate nanoparticle transport to oocytes, and offer insights on mechanisms that may lead to alterations in fish embryos.

该奖项由2009年《美国复苏和再投资法案》(公法111-5)资助。不断增长的证据表明，许多纳米材料通过鳃或消化途径进入鱼类物种。一旦内化，纳米材料有可能进入血流，在那里它们可能会对包括卵巢在内的内部器官构成风险。新出现的文献和目前应用中的初步证据表明，纳米材料可能能够进入卵母细胞，潜在地影响胚胎发育。有必要了解使纳米材料能够运输到卵母细胞的基本机制，以及随后自然生物过程的变化。目前人们了解到，纳米材料的生理行为在很大程度上受到吸附的蛋白质层的影响，吸附的蛋白质层在接触生物介质几秒钟后就会形成。活跃繁殖的雌性鱼类血清中卵黄前体蛋白卵黄蛋白原含量很高(高达20毫克/毫升)。这种蛋白质是一种大的磷脂糖蛋白，能够通过各种途径与纳米材料结合，包括高负电荷区和高疏水性区域。卵黄蛋白原自然针对发育中的卵母细胞，并通过卵黄蛋白原特异性受体输入到这些卵母细胞中。该项目的总体假设是，卵黄原蛋白结合的纳米材料将被引入发育中的卵母细胞的卵黄颗粒中，最终与溶酶体融合。此外，某些纳米材料一旦到达这个隔间，就会溶解并渗入细胞，对发育中的胚胎造成毒性。为了解决这些假设，研究人员有三个具体的目标：(1)确定表面修饰对鱼类卵巢培养物中纳米材料的吸收、细胞分布和保留的影响。(2)研究了雄性和雌性傻瓜鱼血浆对纳米材料表面蛋白质涂层的影响。(3)测定纳米粒在体内的分布和生殖毒性。为了实现这些目标，他们将使用模型发光纳米颗粒，即染料掺杂的二氧化硅和经过精心选择的尺寸分布和表面属性的量子点，这将使详细分析这些材料在体外和体内的运动成为可能。这项拟议的研究将为纳米颗粒特性在它们转移到卵母细胞中的作用提供关键的见解。预计这一信息对于开发鱼类纳米毒性预测模型以及减轻纳米材料暴露在环境中的潜在不利后果的方法具有非常重要的意义。对研究生、本科生和高中生进行粒子科学、工程学和生物学之间的跨学科培训将是研究计划的重要组成部分。东南研究生教育和教授联盟(SEAGEP)、本科生相关研究经验(REU)计划以及佛罗里达大学现有的少数族裔奖学金计划将被利用，以确保研究计划的多样性。研究成果将在同行评议的期刊上发表，并在国家和国际会议上传播。研究人员还计划与亚利桑那州立大学的社会纳米技术中心和莱斯的ICON等国家纳米技术资源合作，向科学和教育界以及整个社会传播研究成果。学生还将参加公共外展项目(例如，UF的工程博览会)，为他们提供一个渠道，让他们参与普通公众对纳米科学的兴奋和环境管理的重要性。UF的格雷厄姆公共政策中心将参与确保将研究结果适当地传达给政策制定者。这项拟议的研究将提供有关纳米颗粒转运到卵母细胞的中介因素的基础知识，并为可能导致鱼类胚胎变化的机制提供见解。