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Kaitlin Ulin Master's Thesis.pdf (1.85 MB)

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Genomic perspectives on the propagation of freshwater mussels

Ulin, Kaitlin
http://rave.ohiolink.edu/etdc/view?acc_num=osu1714929564381821

Abstract Details

2024, Master of Science, Ohio State University, Evolution, Ecology and Organismal Biology.
Artificial propagation, called in vitro propagation, has been commonly used to metamorphose juvenile freshwater mussels without the need of a fish host, making it a useful tool for conservation biology. To improve our success of in vitro propagation, we must first expand our current limited knowledge of the larval development of mussels. The key to this is understanding what nutrition is needed for the successful development and propagation of healthy juvenile mussels. In this study, we compare media of various compositions to assess the growth of larval mussels. Growth is evaluated via analysis of the transcriptome, which can reveal internal processes happening within an organism. This study combines the accessibility of in vitro propagation with gene expression analysis of larval growth between growth media of different compositions with the goal of improving in vitro juvenile mussel propagation. Glochidia were extracted from adult Lampsilis siliquoidea mussels, pooled, and separated into three media groups: Leibovitz's L-15 Medium (L-15), Medium 199 (M199), and M199 with 50μL of lipids from concentrate (M199 + lipids). Glochidia developed in an incubator until signs of metamorphosis were present. Percent metamorphosis in the newly metamorphosed juveniles propagated in each of the three media was calculated and compared. Juveniles were collected from each dish, rinsed with sterile ultrapure water, and snap frozen in liquid nitrogen. RNA was extracted from the samples and sequenced on the Illumina NovaSeq 6000 sequencer with output as 100-base-pair paired-end reads. Assembly of the de novo transcriptome was performed and differentially expressed transcripts between the three groups were identified. Percent metamorphosis was significantly different between the L-15 medium and the M199 medium, but not different between the M199 medium and M199 medium with the addition of lipids. Differential expression was detected in 22,066 transcripts between L-15 and M199, and in 22,435 transcripts between M199 and lipid supplemented M199. Proteins corresponding to these differentially expressed transcripts were identified. Among the differentially expressed transcripts present between L-15 and M199, most transcripts were downregulated in L-15. Differentially expressed transcripts between M199 and M199 with lipids were also mostly downregulated in M199 with lipids. The results from this study indicate that both L-15 and M199 with lipids media were a less stressful environment for L. siliquoidea glochidia compared to the standard M199 medium. However, glochidia propagated in L-15 medium showed downregulation of metabolic processes, indicating that these glochidia may be less metabolically active than those propagated in M199 medium. The significant difference in metamorphosis percentage between L-15 and M199 complements the conclusion that less growth was occurring in the L-15 medium. The results from this study also indicate that the addition of lipids to M199 medium for propagation resulted in dramatic changes in transcript regulation when compared to standard M199 medium. Many transcripts relating to metabolism were downregulated in M199 with lipids. Interestingly, no transcripts directly related to lipid metabolism were upregulated in M199 with lipids. Overall, these results and their interpretation indicate that L-15 and M199 with lipids may not be ideal for the development of L. siliquoidea glochidia, as many important cellular mechanisms were downregulated in both media compared to the standard M199.
Ieva Roznere (Advisor)
Jim Hood (Committee Member)
Lindsey Bruckerhoff (Committee Member)
Meg Daly (Advisor)
100 p.
Conservation; Organismal Biology; Zoology
freshwater mussels; unionid; transcriptomics; in vitro; Lampsilis siliquoidea; omics

Recommended Citations

Citations

  • Ulin, K. (2024). Genomic perspectives on the propagation of freshwater mussels [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1714929564381821

    APA Style (7th edition)

  • Ulin, Kaitlin. Genomic perspectives on the propagation of freshwater mussels. 2024. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1714929564381821.

    MLA Style (8th edition)

  • Ulin, Kaitlin. "Genomic perspectives on the propagation of freshwater mussels." Master's thesis, Ohio State University, 2024. http://rave.ohiolink.edu/etdc/view?acc_num=osu1714929564381821

    Chicago Manual of Style (17th edition)

osu1714929564381821
167
© 2024, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.