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Looking at the metabolic consequences of the colchicine-based in vivo autophagic flux assay.

Autophagy (2016-02-24)
Iban Seiliez, Ikram Belghit, Yujie Gao, Sandrine Skiba-Cassy, Karine Dias, Marianne Cluzeaud, Didier Rémond, Nordine Hafnaoui, Bénédicte Salin, Nadine Camougrand, Stéphane Panserat
ABSTRACT

Monitoring autophagic flux in vivo or in organs remains limited and the ideal methods relative to the techniques possible with cell culture may not exist. Recently, a few papers have demonstrated the feasibility of measuring autophagic flux in vivo by intraperitoneal (IP) injection of pharmacological agents (chloroquine, leupeptin, vinblastine, and colchicine). However, the metabolic consequences of the administration of these drugs remain largely unknown. Here, we report that 0.8 mg/kg/day IP colchicine increased LC3-II protein levels in the liver of fasted trout, supporting the usefulness of this drug for studying autophagic flux in vivo in our model organism. This effect was accompanied by a decrease of plasma glucose concentration associated with a fall in the mRNA levels of gluconeogenesis-related genes. Concurrently, triglycerides and lipid droplets content in the liver increased. In contrast, transcript levels of β-oxidation-related gene Cpt1a dropped significantly. Together, these results match with the reported role of autophagy in the regulation of glucose homeostasis and intracellular lipid stores, and highlight the importance of considering these effects when using colchicine as an in vivo "autophagometer."

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Sigma-Aldrich
1,2-Propanediol, puriss. p.a., ACS reagent, ≥99.5% (GC)
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Oil Red O, certified by the Biological Stain Commission