Presentation Type

Poster

Start Date

5-8-2024 11:00 AM

End Date

5-8-2024 1:00 PM

Subjects

Killifishes

Advisor

Jason Podrabsky

Student Level

Undergraduate

Abstract

The killifish Austrofundulus limnaeus is an extremophile native to small temporary ponds of Venezuela. Normal embryonic development of A. limnaeus is segmented into up to three stages of metabolic depression (diapause), and both developing and diapausing embryos can survive long bouts of the complete absence of oxygen (anoxia) by using anaerobic metabolic pathways. While there has been much focus on how mutations in DNA drive evolution, environmental stress can cause heritable changes to histone post-translational modifications (hPTMs) in multiple species. However, this evolutionary role of hPTMs has never been studied in A. limnaeus. Additionally, the possible evolutionary role of extracellular lactate via histone lactylation has never been studied in any species to our knowledge. To study these phenomena at the cellular level, we will force PSU-AL-WS40NE cells to undergo artificial selection. PSU-AL-WS40NE, is a neuroepithelial cell line that was isolated from embryonic A. limnaeus tissue explant. These cells can survive for 49 days without oxygen while accumulating lactate at a lower rate than mammalian cells. In mammalian cells, 25mM extracellular sodium lactate is sufficient to increase histone lactylation. However, our preliminary data has indicated that 30mM-100mM sodium lactate is not a strong selection pressure, while similar concentrations of lactic acid are lethal to the vast majority of cells. This suggests acidity may be an important component for selection and preconditioning. Therefore, repeated lactic acid exposure will be used as an artificial selection pressure to derive cells with a greater tolerance to high concentrations of extracellular lactate, and therefore potentially greater anoxia tolerance. Should a new cell line be developed, changes in hPTMs will be quantified to assess their potential role in driving changes in gene expression.

Creative Commons License or Rights Statement

Creative Commons Attribution-Noncommercial 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License

Included in

Biology Commons

Share

COinS
 
May 8th, 11:00 AM May 8th, 1:00 PM

Histone Changes as a Response to Lactic Acid Selection in Cells of Austrofundulus limnaeus

The killifish Austrofundulus limnaeus is an extremophile native to small temporary ponds of Venezuela. Normal embryonic development of A. limnaeus is segmented into up to three stages of metabolic depression (diapause), and both developing and diapausing embryos can survive long bouts of the complete absence of oxygen (anoxia) by using anaerobic metabolic pathways. While there has been much focus on how mutations in DNA drive evolution, environmental stress can cause heritable changes to histone post-translational modifications (hPTMs) in multiple species. However, this evolutionary role of hPTMs has never been studied in A. limnaeus. Additionally, the possible evolutionary role of extracellular lactate via histone lactylation has never been studied in any species to our knowledge. To study these phenomena at the cellular level, we will force PSU-AL-WS40NE cells to undergo artificial selection. PSU-AL-WS40NE, is a neuroepithelial cell line that was isolated from embryonic A. limnaeus tissue explant. These cells can survive for 49 days without oxygen while accumulating lactate at a lower rate than mammalian cells. In mammalian cells, 25mM extracellular sodium lactate is sufficient to increase histone lactylation. However, our preliminary data has indicated that 30mM-100mM sodium lactate is not a strong selection pressure, while similar concentrations of lactic acid are lethal to the vast majority of cells. This suggests acidity may be an important component for selection and preconditioning. Therefore, repeated lactic acid exposure will be used as an artificial selection pressure to derive cells with a greater tolerance to high concentrations of extracellular lactate, and therefore potentially greater anoxia tolerance. Should a new cell line be developed, changes in hPTMs will be quantified to assess their potential role in driving changes in gene expression.