Extended indirect calorimetry with isotopic CO2 sensors for prolonged and continuous quantification of exogenous vs. total substrate oxidation in mice

Abstract

Indirect calorimetry (InCa) estimates whole-body energy expenditure and total substrate oxidation based on O₂ consumption and CO₂ production, but does not allow for the quantification of oxidation of exogenous substrates with time. To achieve this, we incorporated ¹³CO₂ and ¹²CO₂ gas sensors into a commercial InCa system and aimed to demonstrate their performance and added value. As a performance indicator, we showed the discriminative oscillations in ¹³CO₂ enrichment associated with food intake in mice fed diets containing naturally low (wheat) vs high (maize) ¹³C enrichment. To demonstrate the physiological value, we quantified exogenous vs total carbohydrate and fat oxidation continuously, in real time in mice varying in fat mass. Diet-induced obese mice were fed a single liquid mixed meal containing ¹³C-isotopic tracers of glucose or palmitate. Over 13 h, ~70% glucose and ~48% palmitate ingested were oxidised. Exogenous palmitate oxidation depended on body fat mass, which was not the case for exogenous glucose oxidation. We conclude that extending an InCa system with ¹³CO₂ and ¹²CO₂ sensors provides an accessible and powerful technique for real-time continuous quantification of exogenous and whole-body substrate oxidation in mouse models of human metabolic physiology.