Determination of the type and quantity of sialic acid in the egg jelly coat of the sea urchin Paracentrotus lividus using capillary LC-ESI-MS/MS.

Sialic acid is a terminal sugar of carbohydrate chains that participates in numerous biological events. Recent studies have explored the mechanism of carbohydrate-mediated fertilisation to understand the biochemistry of fertilisation, although the type and quantity of sialic acid and the role of sialic acid during fertilisation remain unknown. Echinoderm fertilisation in particular has been studied extensively, yet our understanding of the mechanisms of carbohydrate-mediated fertilisation and the role of sialic acid remains incomplete. In this study, we characterised the sialic acid types in the egg jelly coat of the sea urchin, Paracentrotus lividus, using the sensitive analytical system capillary liquid chromatography electro-spray ionisation tandem mass spectrometry (capLC-ESI-MS/MS). First, we isolated the egg jelly coat and released its sialic acid using acid treatment. These sialic acids were derivatised with 1,2-diamino-4,5-methylenediaoxy-benzene dihydrochloride (DMB) and injected into the capLC-ESI-MS/MS system. When compared with standards, we identified twelve different types of sialic acid according to their retention times and collision-induced dissociation fragments. The mass spectral data revealed that Neu5Gc, Neu5Ac, Neu5GcS, and Neu5Gc9Ac were the predominant types of sialic acid in the sea urchin jelly coat, with Neu5Gc being the most abundant. Other types of sialic acid detected included Neu5AcS, Neu5Gc7,9Ac2, Neu5,9Ac2, Neu5Gc8Ac, Neu5Gc7Ac, Neu5,7Ac2, Neu5Gc8,9Ac2, and Neu5,8Ac2. The types and quantities of sialic acid that we detected in the egg jelly coat will aid in the discovery of new sialic acid-specific receptors on the sperm membrane.

Changes in sialic acid expression in the lung during intrauterine development of the human fetus.

Sialic acid is a component of glycoproteins that influences enzymatic and receptor functions of cells. During proliferation and differentiation of tissues, sialic acid can serve as a recognition determinant in intercellular communication and interactions of cells with the extracellular matrix. In the present study, sialic acid expression in relation to developmental maturity of the lung has been studied. We analyzed 12 necroptic lung specimens from foetuses of different gestational ages from the 15th week to the neonate. Sections were stained histochemically using 3 lectins specific for sialic acid: Tritrichomonas mobilensis lectin (TML), specific for sialic acid without linkage preference, Sambucus nigra agglutinin (SNA), specific for alpha2,6-linked sialic acid, and Maackia amurensis leucoagglutinin (MAL), specific for alpha2,3-linked sialic acid. MAL positivity dominated over SNA positivity showing prevalence of alpha2,3-linked sialic acids to be homogeneously distributed in the lung at the canalicular stage of development. In more mature lungs, well-differentiated bronchial epithelium showed strong sialic acid expression of both linkages. Sialic acid with alpha2,6 linkage dominated in vascular endothelium. Our results showed a slight decrease in sialic acid expression in lungs with gestational age to a relative minimum before birth. Lectin staining of mature lung tissue showed intense sialic acid expression in alveolar epithelial type II cells. Changes in expression of specific sialic acids during differentiation of the lungs may be useful as marker of the degree of maturity of the foetus.