Characterization of Histone Modifications in Response to Anoxia in Killifish Cells

Presentation Type

Poster

Start Date

5-8-2024 11:00 AM

End Date

5-8-2024 1:00 PM

Subjects

Embryology--Congresses, Genetics

Advisor

Jason Podrabsky

Student Level

Doctoral

Abstract

Embryos of the annual killifish Austrofundulus limnaeus have the greatest tolerance to anoxia of all vertebrates, making them ideal to study the cellular mechanisms necessary for anoxia tolerance. This tolerance is supported by the ability of embryos to arrest development in diapause as part of their normal development and to enter into a state of anoxia-induced quiescence even during active development. While key RNAs have been described, the regulatory mechanisms that control anoxic gene expression have not. Gene expression is regulated by a variety of mechanisms, including alteration of chromatin through histone modification. Preliminary data in WS 36 embryos (anoxia LT50 of 65 days) identified 626 peptides across fifteen distinct histone proteins, representative of H1, H2A, H2B, and H3 isoforms. Since whole embryo patterns of histone modifications are an aggregate of multiple cell types, we are repeating this work using WS40NE cells, a neuroepithelial cell line isolated from embryonic A. limnaeus tissue explant that can survive anoxia for 49 days. Mass spectrometry based proteomics will be used to quantify histone modifications that occur in anoxic, anoxia-recovered, and normoxic WS40NE cells and compared to modifications in WS 36 embryos. Understanding patterns of histone modification in response to anoxia will provide insight into how cells can utilize histone posttranslational modifications to alter gene expression and support stress tolerance. This work is partly funded by NSF (2025832, 2209383).

Creative Commons License or Rights Statement

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

Persistent Identifier

https://archives.pdx.edu/ds/psu/41867

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

Characterization of Histone Modifications in Response to Anoxia in Killifish Cells

Embryos of the annual killifish Austrofundulus limnaeus have the greatest tolerance to anoxia of all vertebrates, making them ideal to study the cellular mechanisms necessary for anoxia tolerance. This tolerance is supported by the ability of embryos to arrest development in diapause as part of their normal development and to enter into a state of anoxia-induced quiescence even during active development. While key RNAs have been described, the regulatory mechanisms that control anoxic gene expression have not. Gene expression is regulated by a variety of mechanisms, including alteration of chromatin through histone modification. Preliminary data in WS 36 embryos (anoxia LT50 of 65 days) identified 626 peptides across fifteen distinct histone proteins, representative of H1, H2A, H2B, and H3 isoforms. Since whole embryo patterns of histone modifications are an aggregate of multiple cell types, we are repeating this work using WS40NE cells, a neuroepithelial cell line isolated from embryonic A. limnaeus tissue explant that can survive anoxia for 49 days. Mass spectrometry based proteomics will be used to quantify histone modifications that occur in anoxic, anoxia-recovered, and normoxic WS40NE cells and compared to modifications in WS 36 embryos. Understanding patterns of histone modification in response to anoxia will provide insight into how cells can utilize histone posttranslational modifications to alter gene expression and support stress tolerance. This work is partly funded by NSF (2025832, 2209383).