A test of alternate models for increased tissue nitrogen isotope ratios during fasting in hibernating arctic ground squirrels

Abstract

We describe two models explaining the increase in tissue nitrogen isotope ratios (δ(15)N) that occurs during fasting in animals. The catabolic model posits that protein breakdown selectively removes the lighter isotope of nitrogen ((14)N) from catabolized tissues, causing an increase in the proportion of heavy nitrogen isotope ((15)N). The anabolic model posits that protein synthesis during fasting results in elevated δ(15)N values, as the unreplaced loss of (14)N to urea results in a higher proportion of (15)N in plasma amino acids used for protein synthesis. We effected a range of lean mass loss in arctic ground squirrels (Urocitellus parryii) fasting during hibernation and then collected organ and muscle tissues for analysis of δ(15)N values. The catabolic model predicts increased δ(15)N values in both liver and muscle, as these tissues undergo significant catabolism during hibernation. The anabolic model predicts no change in muscle, but an increase in δ(15)N values in liver, which has high levels of protein synthesis during euthermic phases of hibernation. We found a significant increase in liver δ(15)N values and no change in muscle δ(15)N values with lean mass loss, which supports the anabolic model. Heart, small intestine and brown adipose tissue also showed an increase in δ(15)N values, indicating protein synthesis in these organ tissues during hibernation. Urine was 3.8% lighter than plasma, and both urine and plasma increased in δ(15)N values with lean mass loss. This study helps clarify the mechanisms causing δ(15)N change during nutritional stress, thus increasing its utility for physiological research and reconciling previously contradictory results.