Natural Isotopic Signatures of Variations in Body Nitrogen Fluxes: A Compartmental Model Analysis

Body tissues are generally 15N-enriched over the diet, with a discrimination factor (Δ15N) that varies among tissues and individuals as a function of their nutritional and physiopathological condition. However, both 15N bioaccumulation and intra- and inter-individual Δ15N variations are still poorly understood, so that theoretical models are required to understand their underlying mechanisms. Using experimental Δ15N measurements in rats, we developed a multi-compartmental model that provides the first detailed representation of the complex functioning of the body's Δ15N system, by explicitly linking the sizes and Δ15N values of 21 nitrogen pools to the rates and isotope effects of 49 nitrogen metabolic fluxes. We have shown that (i) besides urea production, several metabolic pathways (e.g., protein synthesis, amino acid intracellular metabolism, urea recycling and intestinal absorption or secretion) are most probably associated with isotope fractionation and together contribute to 15N accumulation in tissues, (ii) the Δ15N of a tissue at steady-state is not affected by variations of its P turnover rate, but can vary according to the relative orientation of tissue free amino acids towards oxidation vs. protein synthesis, (iii) at the whole-body level, Δ15N variations result from variations in the body partitioning of nitrogen fluxes (e.g., urea production, urea recycling and amino acid exchanges), with or without changes in nitrogen balance, (iv) any deviation from the optimal amino acid intake, in terms of both quality and quantity, causes a global rise in tissue Δ15N, and (v) Δ15N variations differ between tissues depending on the metabolic changes involved, which can therefore be identified using simultaneous multi-tissue Δ15N measurements. This work provides proof of concept that Δ15N measurements constitute a new promising tool to investigate how metabolic fluxes are nutritionally or physiopathologically reorganized or altered. The existence of such natural and interpretable isotopic biomarkers promises interesting applications in nutrition and health.
Author Summary Body proteins ensure vital functions, and their constancy is maintained through the tight coordination of many nitrogen metabolic fluxes, but our understanding of how this flux system is regulated, and sometimes dysregulated, remains fragmentary and incomplete. Besides, body tissues are generally naturally enriched in the heavier stable nitrogen isotope (15N) over the diet: this 15N bioaccumulation (Δ15N) varies depending on tissues and metabolic orientations, likely as the result of isotope effects associated to some metabolic pathways. We used a novel approach, combining multi-tissue Δ15N measurements and their analysis using modeling, to understand how body Δ15N values relate to nitrogen fluxes. The multi-tissue model we have developed provides a clearer understanding of the metabolic processes that generate isotopic fractionation, and of how tissue Δ15N values are modulated in response to changes in the body distribution of specific nitrogen fluxes. We show that Δ15N values tend to rise when the amino acids intake does not optimally fit the metabolic demand, and that Δ15N values constitute natural and interpretable signatures of nutritionally-induced variations in nitrogen fluxes. This approach constitutes a new promising tool to investigate how nitrogen metabolism is nutritionally or physiopathologically reorganized or altered, and promises interesting applications in many areas (nutrition, pathology, ecology, paleontology, etc).