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劳动力的存在，这些个人是STEM领域代表性不足的群体的成员。CREST-PRP奖项表彰具有巨大潜力的研究人员，并支持他们的研究经验，以拓宽他们的视野，促进跨学科互动，并为CREST-PRP学者在科学界担任领导职位做好准备。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CREST波多黎各环境神经科学中心的一名附属研究人员提交，将调查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。这项跨学科的研究将集中于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了解。此外，通过科学专家的积极指导和参与这一经历提供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研究生涯做准备。他还将参与几项外展活动，为STEM领域代表性不足的群体的学生树立榜样，激励和激励下一代环境神经科学家的更广泛参与。该项目的长期目标是介绍和探索Biomphalaria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。接触镉和锰对可存活卵子生产的影响将提供它们对繁殖力影响的一种衡量标准。利用免疫组织化学和一种新的原位杂交方法，将在蛋白质和mRNA水平上检测污染物暴露对控制两性钉螺雄性和雌性行为的神经肽表达的影响。将使用CRISPR/Cas9基因敲除和敲除方法来研究单个肽能系统对生殖生理和行为的贡献。神经转录组将被用来检测接触镉和锰是否会改变控制男性和女性生殖行为的多肽的表达。将使用HCR(杂交链式反应)方案进行原位杂交，以在暴露于污染物前后在转录水平上测量这些多肽的mRNA表达。最后，CRISPR/Cas9介导的基因组修改将对调节生殖行为的特定神经基因进行。通过这项工作增加对污染物对光肩星天牛神经肽和生殖行为影响的了解，可以改进环境保护、人类健康、疾病控制和保护工作的战略。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.