Salinity Effects on Lytechinus variegatus Development
By: Davina Kara, Isabella Laske, Shane Webb and Margaret Smith
Abstract
Climate change, through altered patterns of temperature, rainfall and evaporation, is affecting ocean salinity in some areas. These changes have a direct impact on the growth and development of many marine organisms with mineral skeletons, including sea urchins. Among other changes, atypical salinity can interrupt arm growth in Lytechinus variegatus larvae, likely through impacts on developmental gene expression. Importantly, the sea urchin gene regulatory network underlying skeletal arm development is well known. We here wish to establish the impact of altered salinity on L. variegatus arm development using qPCR to measure gene expression differences. To do this, we cultured L. variegatus larvae for 72 hours (the window for arm development) at three salinities: 24 ppt, 25 ppt, and our control, 27 ppt. We then isolated RNA from pooled samples of these larvae. We are currently assessing primers for qPCR. To date we have evaluated 15 primer sets, 12 of which we designed, and three of which we found in the literature. All three published primer sets amplified. Two sets, Setmar (housekeeping gene) and SM30, amplified a single band of correct sequence identity, and the third amplified an erroneous product. Of the primers we designed, five failed to amplify, six amplified multiple bands and thus are not appropriate for qPCR, and one amplified a single band that was not verified as the correct product through sequencing. These outcomes highlight the challenges in optimizing qPCR primers for specific gene amplification. Our next steps include continuing to optimize primers and conducting qPCR using the two sets of primers that successfully amplified.
Language: English
Published on: Mar 20, 2025
Published by: University of North Georgia
In partnership with: Paradigm Publishing Services
Keywords:
© 2025 Davina Kara, Isabella Laske, Shane Webb, Margaret Smith, published by University of North Georgia
This work is licensed under the Creative Commons Attribution 4.0 License.