title: In situ visualization of carbonylation and its co-localization with proteins, lipids, DNA and RNA in Caenorhabditis elegans.
abstract: All key biological macromolecules are susceptible to carbonylation - an irreparable oxidative damage with deleterious biological consequences. Carbonyls in proteins, lipids and DNA from cell extracts have been used as a biomarker of oxidative stress and aging, but formation of insoluble aggregates by carbonylated proteins precludes quantification. Since carbonylated proteins correlate with and become a suspected cause of morbidity and mortality in some organisms, there is a need for their accurate quantification and localization. Using appropriate fluorescent probes, we have developed an in situ detection of total proteins, DNA, RNA, lipids and carbonyl groups at the level of the whole organism. In C. elegans, we found that after UV irradiation carbonylation co-localizes mainly with proteins and, to a lesser degree, with DNA, RNA and lipids. The method efficiency was illustrated by carbonylation induction assessment over 5 different UV doses. The procedure enables the monitoring of carbonylation in the nematode C. elegans during stress, aging and disease along its life cycle including the egg stage.
authors.affiliation: Institut de radioprotection et de sûreté nucléaire, Cadarache, 13115 Saint Paul lez Durance cedex, France; Mediterranean Institute for Life Sciences, Mestrovicevo Setaliste 45, 21000 Split, Croatia.
authors.forename: Mira
authors.initials: M
authors.lastname: Kuzmic
authors.affiliation: CEA, BIAM, Lab Biol Develop Plantes, Saint-Paul-lez-DurIncreased carbonylation, protein aance F-13108, France; CNRS, UMR 7265 Biol Veget & Microbiol Environ, Saint-Paul-lez-Durance F-13108, France; Aix Marseille Université, BVME UMR7265, Marseille F-13284, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Javot
authors.affiliation: Institut de radioprotection et de sûreté nucléaire, Cadarache, 13115 Saint Paul lez Durance cedex, France.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Bonzom
authors.affiliation: Institut de radioprotection et de sûreté nucléaire, Cadarache, 13115 Saint Paul lez Durance cedex, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Lecomte-Pradines
authors.affiliation: Mediterranean Institute for Life Sciences, Mestrovicevo Setaliste 45, 21000 Split, Croatia.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Radman
authors.affiliation: Institut de radioprotection et de sûreté nucléaire, Cadarache, 13115 Saint Paul lez Durance cedex, France.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Garnier-Laplace
authors.affiliation: Institut de radioprotection et de sûreté nucléaire, Cadarache, 13115 Saint Paul lez Durance cedex, France. Electronic address: sandrine.frelon@irsn.fr.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Frelon
mesh_terms: D000818:Animals; D001896:Boron Compounds / Q000737:chemistry; D017173:Caenorhabditis elegans / Q000254:growth & development / Q000378:metabolism / Q000528:radiation effects* / Q000648:ultrastructure; D029742:Caenorhabditis elegans Proteins / Q000032:analysis* / Q000737:chemistry / Q000528:radiation effects; D003374:Coumarins / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D005456:Fluorescent Dyes / Q000737:chemistry; D006834:Hydrazines / Q000737:chemistry; D007211:Indoles / Q000737:chemistry; D008055:Lipids / Q000032:analysis* / Q000737:chemistry / Q000528:radiation effects; D009930:Organic Chemicals / Q000737:chemistry; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D050050:Protein Carbonylation; D012313:RNA / Q000032:analysis* / Q000737:chemistry / Q000528:radiation effects; D014466:Ultraviolet Rays
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D029742:Caenorhabditis elegans Proteins; D003374:Coumarins; D005456:Fluorescent Dyes; D006834:Hydrazines; D007211:Indoles; D008055:Lipids; D009930:Organic Chemicals; C098798:SYBR Green II; C446799:dinitrophenylhydrazine; C007293:DAPI; D012313:RNA; D004247:DNA
keywords: Caenorhabditis elegans; DNA carbonylation; Fluorescent probes; In situ carbonylation; Lipid carbonylation; Protein carbonylation; RNA carbonylation