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

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.