Derivation and characterization of a California grunion (Leuresthes tenuis) embryonic cell line

An immortalized cell line was established from newly hatched embryos of the California grunion (Leuresthes tenuis), a silverside fish, to generate a tool for mechanistic studies explaining the effect of environmental stressors on proteome dynamics. In this oviparous species, fertilization occurs terrestrially, and embryos develop in beach sand until environmentally-cued hatching. Grunion eggs and embryos are subject to multiple environmental stressors including changes in salinity, desiccation, heat, and pollutant runoff, which can reduce fertilization, embryonic development, and hatching success [1–3]. Cell lines derived from grunion embryo primary tissue explants thus provide a valuable tool for high-throughput reverse genetics (e.g., high-throughput enhancer trap assays, knock-out studies, and targeted mutagenesis) and proteomics approaches to enable mechanistic explanations of cellular and molecular responses to environmental stressors. Two parallel cell lines were derived from the same whole grunion embryonic tissue and have been maintained now over 10 passages at a 1:6 splitting ratio. Both cell lines display an epithelial-like phenotype (cuboidal, monolayer, optically dense). A time course experiment was conducted to document effects of cell immortalization on the proteome of both cell lines sampled at each passage. Data-dependent mass spectrometry acquisition of peptide spectra and de novo sequencing were used to annotate spectral libraries with protein identification information. Spectral libraries were then used to construct and validate target sets that enable quantitation of proteome dynamics with data-independent acquisition (cDIA) and Skyline. Passage-dependent changes in proteome dynamics were identified by cDIA target set quantitation with Skyline and correspond to morphological changes in cell types over time. Support or Funding Information Supported by NSF Grant IOS-1355098. Fibroblast and epithelial-like cells derived from grunion after the first passage (A) and third passage (B). This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.