Impact of anesthesia and euthanasia on metabolomics of mammalian tissues: studies in a C57BL/6J mouse model.

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TitleImpact of anesthesia and euthanasia on metabolomics of mammalian tissues: studies in a C57BL/6J mouse model.
Publication TypeJournal Article
Year of Publication2015
AuthorsOvermyer KA, Thonusin C, Qi NR, Burant CF, Evans CR
JournalPLoS One
Volume10
Issue2
Paginatione0117232
Date Published2015
ISSN1932-6203
KeywordsAnesthesia, Anesthetics, Dissociative, Anesthetics, Inhalation, Animals, Euthanasia, Animal, Humans, Isoflurane, Ketamine, Metabolome, Metabolomics, Mice, Organ Specificity
Abstract

A critical application of metabolomics is the evaluation of tissues, which are often the primary sites of metabolic dysregulation in disease. Laboratory rodents have been widely used for metabolomics studies involving tissues due to their facile handing, genetic manipulability and similarity to most aspects of human metabolism. However, the necessary step of administration of anesthesia in preparation for tissue sampling is not often given careful consideration, in spite of its potential for causing alterations in the metabolome. We examined, for the first time using untargeted and targeted metabolomics, the effect of several commonly used methods of anesthesia and euthanasia for collection of skeletal muscle, liver, heart, adipose and serum of C57BL/6J mice. The data revealed dramatic, tissue-specific impacts of tissue collection strategy. Among many differences observed, post-euthanasia samples showed elevated levels of glucose 6-phosphate and other glycolytic intermediates in skeletal muscle. In heart and liver, multiple nucleotide and purine degradation metabolites accumulated in tissues of euthanized compared to anesthetized animals. Adipose tissue was comparatively less affected by collection strategy, although accumulation of lactate and succinate in euthanized animals was observed in all tissues. Among methods of tissue collection performed pre-euthanasia, ketamine showed more variability compared to isoflurane and pentobarbital. Isoflurane induced elevated liver aspartate but allowed more rapid initiation of tissue collection. Based on these findings, we present a more optimal collection strategy mammalian tissues and recommend that rodent tissues intended for metabolomics studies be collected under anesthesia rather than post-euthanasia.

DOI10.1371/journal.pone.0117232
Alternate JournalPLoS ONE
PubMed ID25658945
PubMed Central IDPMC4319778
Grant ListK25 DK092558 / DK / NIDDK NIH HHS / United States
K25 DK092558 / DK / NIDDK NIH HHS / United States
P30 DK020572 / DK / NIDDK NIH HHS / United States
P30 DK089503 / DK / NIDDK NIH HHS / United States
P30 DK089503 / DK / NIDDK NIH HHS / United States
R01 DK077200 / DK / NIDDK NIH HHS / United States
U24 DK097153 / DK / NIDDK NIH HHS / United States
U24 DK097153 / DK / NIDDK NIH HHS / United States