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SILAC zebrafish for quantitative analysis of protein turnover and tissue regeneration.

Journal article
Authors Ann Brinkmalm-Westman
Alexandra Abramsson
Josef Pannee
Chen Gang
Mikael K Gustavsson
Malin von Otter
Kaj Blennow
Gunnar Brinkmalm
Hermann Heumann
Henrik Zetterberg
Published in Journal of proteomics
Volume 75
Issue 2
Pages 425-34
ISSN 1876-7737
Publication year 2011
Published at Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry
Pages 425-34
Language en
Links dx.doi.org/10.1016/j.jprot.2011.08....
Keywords Animal Fins, metabolism, physiology, Animals, Carbon Isotopes, Chromatography, Liquid, Intestines, metabolism, Isotope Labeling, methods, Liver, metabolism, Lysine, metabolism, Regeneration, physiology, Tandem Mass Spectrometry, Zebrafish, metabolism
Subject categories Psychiatry

Abstract

Defective tissue regeneration is thought to contribute to several human diseases, including neurodegenerative disorders, heart failure and various lung diseases. Boosting the regenerative capacity has been suggested a possible therapeutic approach. Methods to metabolically label newly synthesized proteins in vivo with stable isotopic forms of amino acids holds promise for the study of protein turnover and tissue regeneration that are fundamental to the sustained life of all organisms. Here, we used the "stable isotope labeling with amino acids in cell culture" (SILAC) approach to explore normal protein turnover and tissue regeneration in adult zebrafish. The ratio of labeled and unlabeled proteins/peptides in specific organs of zebrafish fed a SILAC diet containing (13)C(6)-labeled lysine was determined by liquid chromatography and tandem mass spectrometry. Labeling was highest in tissues with high regenerative capacity, including intestine, liver, and fin, whereas brain and heart displayed the lowest labeling. Proteins with high degree of labeling were mainly involved in catalytic or transport activity pathways. The technique also verified increased protein synthesis during regeneration of the caudal fin following amputation. This newly developed SILAC zebrafish model constitutes a novel tool to analyze tissue regeneration in an animal model amenable to genetic and pharmacologic manipulation.

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