Myogenic Response to Increasing Concentrations of Ammonia Differs between Mammalian, Avian, and Fish Species: Cell Differentiation and Genetic Study

Abstract

Ammonia is very toxic to the body and has detrimental effects on many different organ systems. Using cultured myoblast cells, we examined ammonia’s effect on myostatin expression, a negative regulator of skeletal muscle growth, and myotube diameters. The objective of this study was to examine how murine, avian, and fish cells respond to increasing levels of ammonia up to 50 mM. The murine myoblast cell line (C2C12), primary chick, and primary tilapia myoblast cells were cultured and then exposed to 10, 25, and 50 mM ammonium acetate, sodium acetate, and an untreated control for 24 h. High levels of ammonia were detrimental to the C2C12 cells, causing increased Myostatin (MSTN) expression and decreased myotube diameters between 10 and 25 mM (p < 0.002). Ammonia at 10 mM continued the positive myogenic response in the chick, with lower MSTN expression than the C2C12 cells and larger myotube diameters, but the myotube diameter at 50 mM ammonium acetate was significantly smaller than those at 10 and 25 mM (p < 0.001). However, chick myotubes at 50 mM were still significantly larger than the sodium acetate-treated and untreated control (p < 0.001). The tilapia cells showed no significant difference in MSTN expression or myotube diameter in response to increasing the concentrations of ammonia. Overall, these results confirm that increasing concentrations of ammonia are detrimental to mammalian skeletal muscle, while chick cells responded positively at lower levels but began to exhibit a negative response at higher levels, as the tilapia experienced no detrimental effects. The differences in ammonia metabolism strategies between fish, avian, and mammalian species could potentially contribute to the differences between species in response to high levels of ammonia. Understanding how ammonia affects skeletal muscle is important for the treatment of muscle wasting observed in liver failure patients.