An Effective Protocol for Proteome Analysis of Medaka (Oryzias latipes ) after Acute Exposure to Ionizing Radiation

Methods and Protocols
Yeni Pérez-GélvezCarl Bergmann

Abstract

All terrestrial organisms are subject to evolutionary pressures associated with natural sources of ionizing radiation (IR). The legacy of human-induced IR associated with energy, weapons production, medicine, and research has changed the distribution and magnitude of these evolutionary pressures. To date, no study has systematically examined the effects of environmentally relevant doses of radiation exposure across an organismal proteome. This void in knowledge has been due, in part, to technological deficiencies that have hampered quantifiable environmentally relevant IR doses and sensitive detection of proteomic responses. Here, we describe a protocol that addresses both needs, combining quantifiable IR delivery with a reliable method to yield proteomic comparisons of control and irradiated Medaka fish. Exposures were conducted at the Savannah River Ecology Laboratory (SREL, in Aiken, SC), where fish were subsequently dissected into three tissue sets (carcasses, organs and intestines) and frozen until analysis. Tissue proteins were extracted, resolved by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE), and each sample lane was divided into ten equal portions. Following in-gel tryptic digestion, peptides r...Continue Reading

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Citations

Dec 22, 2020·International Journal of Radiation Biology·Yeni Natalia C Perez-GelvezCarl W Bergmann
Aug 7, 2021·International Journal of Radiation Biology·Yeni Natalia C Perez-GelvezCarl W Bergmann
Aug 28, 2021·Cancers·Theresa SuckertElke Beyreuther

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Methods Mentioned

BETA
protein assay
Fusion Tribrid
pull off

Software Mentioned

Fusion instrumentation
fasta
Proteomic Pipeline
MASCOT
JPOST
Trans
ProteoIQ

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