Presence of sialic acids in bronchioloalveolar cells and identification and quantification of N-acetylneuraminic and N-glycolylneuraminic acids in the lung

Abstract

The lung, in air-breathing vertebrates, is a tree-like structure composed of branching tubes ending in alveoli and lined by diverse and highly specialized epithelial cells.A dense array of complex and diverse glycoconjugates is present on essentially all animal cell surfaces. Sialic acids are widely allocated at the outermost ends of glycan chains, attached to membrane proteins and lipids below. Due to their abundance and their terminal position in glycans, sialic acids are implicated in many physiological and pathological functions. Although the composition of lung epithelial cell-surface glycans has been studied over the years, it is not yet completely understood. In the present work, we aimed to histochemically localize N-acetylneuraminic acid (Neu5Ac)>N-glycolylneuraminic acid (Neu5Gc) residues on rat bronchioloalveolar epithelial cell surfaces using light microscopy (LM) methods. In lung membranes isolated from adult rat lung homogenates, we also separated, identified and quantified Neu5Ac and Neu5Gc by means of high-performance liquid chromatography (HPLC), and systematically described the optimized HPLC methods used. Sialic acid residues were localized on the surface coat of bronchioloalveolar cells, and the mean quantification of Neu5Ac and Neu5Gc in the adult rat lung homogenates was 12,26 and 2,73 μg/mg prot., respectively, revealing a manifest preponderance of Neu5Ac. A coefficient of variation (CV) of 4,98% and 4,40%, respectively was obtained and an optimal dispersion variability expressed by the SD and the CV was also reported, confirming the efficiency of the methodology.To the best of our knowledge, our group was the first to identify, separate and quantify sialic acids in purified lung membranes.The presence of these residues contributes to a strong anionic shield and may provide an hydrating and protective barrier as well as a repulsive structure that is crucial to lung physiology.