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

CREST博士后研究计划（CREST- prp）为活跃的CREST中心的个人早期职业科学家提供两年的研究、培训和指导经验支持。CREST-PRP奖项的目标是增加STEM领域中代表性不足群体成员的个人在STEM劳动力中的存在。CREST-PRP奖项旨在表彰具有重大潜力的研究人员，并支持他们的研究经验，以拓宽他们的视野，促进跨学科的互动，并为CREST-PRP学者在科学界的领导职位做好准备。研究项目“CREST-PRP：金属对软体动物模型系统繁殖的影响”与CREST-PRP的目标直接一致。该项目由隶属于CREST波多黎各环境神经科学中心的一名研究人员提交，将调查农业和工业应用产生的重金属污染物径流对蜗牛神经生理学的影响。这项跨学科的研究将重点关注并促进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指导和参与这一经历所提供的其他专业发展机会，博士后研究员将进一步为成功的STEM研究生涯做好准备。他还将参与一些外展活动，让他成为STEM领域中代表性不足的群体的学生的榜样，激励和鼓舞下一代环境神经科学家更广泛的参与。该项目的长期目标是介绍和探索生物phalaria神经系统，作为在单个已识别神经元水平上理解生态毒理学影响的模型。暴露于镉和锰对活卵生产的影响将为它们对繁殖力的影响提供一个衡量标准。污染物暴露对控制雌雄同体蜗牛雄性和雌性行为的神经肽表达的影响将在蛋白质和mRNA水平上进行测量，使用免疫组织化学和一种新的原位杂交方法。个体肽能系统对生殖生理和行为的贡献将使用CRISPR/Cas9敲除和敲低方法进行研究。神经转录组将用于检查暴露于镉和锰是否会改变控制雄性和雌性生殖行为的肽的表达。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肽的mRNA表达。最后，CRISPR/Cas9介导的基因组修饰将对调节生殖行为的特定神经基因进行修饰。通过这项工作，我们可以进一步了解污染物对光斑小蠊神经肽和生殖行为的影响，从而改进环境保护、人类健康、疾病控制和保护工作的策略。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.