CREST-PRP: Effects of Metals on Reproduction in a Molluscan Model System

CREST-PRP：金属对软体动物模型系统繁殖的影响

• 批准号: 2329312
• 负责人: Jonathan Crooke Rosado
• 金额: $ 33.88万
• 依托单位: University of Puerto Rico Medical Sciences Campus
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2329312&HistoricalAwards=false
• 关键词: CREST; PRP; Effects; Metals; Reproduction

The CREST Postdoctoral Research Program (CREST-PRP) provides two years of support for research, training, and mentoring experiences for individual early career scientists at active CREST Centers. The goal of the CREST-PRP awards is to increase the STEM workforce presence of individuals who are members of groups underrepresented in STEM fields. CREST-PRP awards recognize investigators with significant potential and support their research experiences to broaden their perspectives, facilitate interdisciplinary interactions, and prepare CREST-PRP scholars for positions of leadership within the scientific community. The research project “CREST-PRP: Effects of Metals on Reproduction in a Molluscan Model System” is in direct alignment with the CREST-PRP goals. Submitted by a researcher affiliated with the CREST Puerto Rico Center for Environmental Neuroscience, the project will investigate the effects of heavy metal contaminant runoff from agricultural and industrial applications on the neurophysiology of snails. This interdisciplinary research study will focus on and advance understanding of the effects of contaminants on individual snail neurons that activate reproductive behaviors. In addition, through active mentoring from scientific experts and participation in other professional development opportunities afforded by this experience, the postdoctoral researcher will further his preparation for a successful STEM research career. He will also engage in several outreach activities, allowing him to serve as a role model for students from groups that are underrepresented in STEM fields, motivating and inspiring broader participation in the next generation of environmental neuroscientists. The long-term goal of this project is to introduce and explore the Biomphalaria nervous system as a model for understanding ecotoxicological impacts at the level of single identified neurons. Effects of exposure to cadmium and manganese on production of viable eggs will provide a measure of their impact on fecundity. Effects of contaminant exposure on expression of neuropeptides that control the male and female behaviors of this hermaphroditic snail will be measured at the protein and mRNA levels, using immunohistochemistry and a novel in situ hybridization method. The contributions of individual peptidergic systems to reproductive physiology and behavior will be examined using CRISPR/Cas9 knockout and knock-down approaches. A neural transcriptome will be used to examine whether exposure to cadmium and manganese alters expression of peptides that control both male and female reproductive behaviors. In-situ hybridizations will be performed using the HCR (Hybridization Chain Reaction) protocol to measure mRNA expression of these peptides at the level of transcription before and after exposure to contaminants. Finally, CRISPR/Cas9 mediated genome modifications will be conducted on specific neural genes that regulate the reproductive behavior. Increased understanding of the effects of contaminants on B. glabrata neuropeptides and reproductive behavior resulting from this work could lead to improved strategies for environmental protection, human health, disease control, and conservation efforts.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

CREST博士后研究计划（Crest-PRP）为Active Crest Centers的个人早期职业科学家提供了两年的研究，培训和心理体验。 CREST-PRP奖项的目标是增加STEM领域中代表性不足的人群的STEM劳动力的存在。 Crest-PRP奖项认可具有巨大潜力并支持其研究经验的调查人员，以扩大其观点，促进跨学科的互动，并为Crest-PRP学者准备科学界的领导地位。研究项目“ Crest-PRP：金属对软体动物模型系统中繁殖的影响”与Crest-PRP目标直接保持一致。该项目由波斯特波多黎各环境神经科学中心提交，该项目将研究农业和工业应用中重金属污染物径流对蜗牛神经生理学的影响。这项跨学科研究将重点关注并提高人们对污染物对激活生殖行为的单个蜗牛神经元的影响的理解。此外，通过科学专家的积极心理和参与这一经验带来的其他专业发展机会，博士后研究人员将进一步为成功的STEM研究职业做准备。他还将从事几项外展活动，使他能够成为来自STEM领域中人数不足的学生的榜样，激励和激发了对下一代环境神经科学家的广泛参与。该项目的长期目标是引入和探索生物掌神经系统，以此作为理解单个水平生态毒理学影响的模型。确定的神经元。暴露于镉和锰对可行鸡蛋产生的影响将衡量它们对繁殖力的影响。污染物暴露对控制这种雌雄同体蜗牛的男性和女性行为的神经肽表达的影响将在蛋白质和mRNA水平上使用免疫组织化学和一种原位杂交方法进行测量。将使用CRISPR/CAS9敲除和敲除方法来检查单个肽能系统复制生理和行为的贡献。神经转录组将用于检查暴露于镉和锰是否会改变控制男性和女性生殖行为的辣椒的表达。将使用HCR（杂交链反应）方案进行原位杂交，以在暴露于污染物之前和之后的转录水平上测量这些辣椒的mRNA表达。最后，将对调节生殖行为的特定神经原体进行CRISPR/CAS9介导的基因组修饰。对这项工作产生的污染物对污染物对葡萄链球菌神经肽的影响以及生殖行为的影响增加可能会导致改进的环境保护，人类健康，疾病控制和保护工作的策略。该奖项反映了NSF的法定任务，并通过该基金会的知识优点和广泛的影响来评估NSF的法定任务，并被认为是值得的支持。