ABSTRACT
OBJECTIVE: Multiplexed metabolic phenotyping systems are available from multiple commercial vendors, and each system includes unique design features. Although expert opinion supports strengths and weaknesses of each design, empirical data from carefully controlled studies to test the biological impact of design differences are lacking. METHODS: Wild-type C57BL/6J mice of both sexes underwent phenotyping in OxyMax (Columbus Instruments International) and Promethion (Sable Systems International) systems located within the same room of a newly constructed animal research facility in a crossover design study. Phenotypes were examined under chow (2920×)-fed conditions and again after 4 weeks of 60% high-fat diet (D12492) feeding. RESULTS: Food intake, physical activity, and respiratory gas exchange data significantly diverged between systems, depending upon sex of animals and diet supplied. Estimates of energy expenditure based on gas exchange in both systems accounted for a fraction of consumed calories that was greater in males than females. CONCLUSIONS: Design differences quantitatively impact the assessment of metabolic end points and thus the qualitative interpretation of various interventions. Importantly, current multiplexed systems remain blind to multiple additional end points, including digestive efficiency and selected forms of energy flux (nitrogenous, anaerobic, etc.), that account for a physiologically and/or pathophysiologically significant fraction of total energy flux.
Subject(s)
Diet, High-Fat/methods , Energy Intake/physiology , Animals , Female , Male , Mice , Mice, Inbred C57BL , PhenotypeABSTRACT
Hallmarks of aging that negatively impact health include weight gain and reduced physical fitness, which can increase insulin resistance and risk for many diseases, including type 2 diabetes. The underlying mechanism(s) for these phenomena is poorly understood. Here we report that aging increases DNA breaks and activates DNA-dependent protein kinase (DNA-PK) in skeletal muscle, which suppresses mitochondrial function, energy metabolism, and physical fitness. DNA-PK phosphorylates threonines 5 and 7 of HSP90α, decreasing its chaperone function for clients such as AMP-activated protein kinase (AMPK), which is critical for mitochondrial biogenesis and energy metabolism. Decreasing DNA-PK activity increases AMPK activity and prevents weight gain, decline of mitochondrial function, and decline of physical fitness in middle-aged mice and protects against type 2 diabetes. In conclusion, DNA-PK is one of the drivers of the metabolic and fitness decline during aging, and therefore DNA-PK inhibitors may have therapeutic potential in obesity and low exercise capacity.
