ABSTRACT
It is known that disialic acids (diSia) are present in the mammalian brain. However, the precise anatomical distribution and the chronology of its expression along life are not well studied yet. It is accepted that the transfer of diSia in the brain is mediated mainly by the enzyme ST8Sia III (α2,8-sialyltransferase III). We studied the expression of diSia glycoepitopes and of the ST8Sia III gene in different structures of the mouse brain at different postnatal stages by immunohistochemistry and real-time polymerase chain reaction, respectively. C57BL/6 mice of different stages were used. Samples of hippocampus, olfactory bulb, cortex and cerebellum were processed for studies of molecular biology and immunohistochemistry. Histological analysis revealed an important decrease in diSia labeling in the senile cerebellum compared with other structures and stages (P ⪠0.001). In concordance with these results, a significant decrease in ST8Sia III gene expression was found in the cerebellum of senile animals (P < 0.001). These results suggest that diSia are constantly expressed but with differential expression in various areas of the mouse central nervous system. On the other hand, the concordance in the decreased expression of ST8Sia III and the diSia epitope in the cerebellum of senile animals suggest a role of diSia in this structure or, inversely, an influence of aging on the expression of diSia in the cerebellum. Further research in that direction could elucidate the roles of diSia in brain function in health and disease.
Subject(s)
Brain Diseases/metabolism , Cerebellum/metabolism , Gene Expression Regulation , Sialic Acids/metabolism , Sialyltransferases/metabolism , Animals , Cerebellum/enzymology , Cerebellum/pathology , Hippocampus/enzymology , Hippocampus/metabolism , Mice , Mice, Inbred C57BL , Olfactory Bulb/enzymology , Olfactory Bulb/metabolism , Organ Specificity , Sialic Acids/biosynthesis , Sialyltransferases/geneticsABSTRACT
Experiments were performed to determine whether sialic acids are expressed in two dermatophytes: Trichophyton rubrum and T. mentagrophytes, similarly to other fungal pathogens. Chemical extraction of mycelia and microconidia followed by high-performance thin-layer chromatography and colorimetric assays were all negative for sialic acid. Incubation of dermatophytes in the presence of Limax flavus agglutinin, specific for sialic acid, was negative in a fluorescence staining test. We have also used other lectins that bind to sialic acid and/or other sugar residues, and these ligands coupled to fluorescein strongly stained these fungi. Such fluorescence staining was not abolished or reduced when fungi were pretreated with sialidase. Different strains of influenza virus which recognize sialic acid residues were also used, but no agglutination of the dermatophytes was observed. Based on these methods, which successfully revealed the presence of sialic acids in other fungal pathogens, we show that these monosaccharides do not occur in both dermatophyte species. Thus, sialic acids do not seem to play a role in the pathogenicity of these fungi.
Subject(s)
Sialic Acids/biosynthesis , Trichophyton/metabolism , Agglutination , Chromatography, High Pressure Liquid , Colorimetry , Fluorescent Antibody Technique, Direct , Lectins/metabolism , Mitochondria/metabolism , Mycelium/chemistry , Mycelium/metabolism , Neuraminidase/pharmacology , Orthomyxoviridae/metabolism , Sialic Acids/analysis , Species Specificity , Trichophyton/drug effectsABSTRACT
The increasing number of reports on the presence of sialic acids in fungi (N-acetyl-, N-glycolyl- and 5,9-N,O-diacetylneuraminic acids) based on direct and indirect evidence warrants the present review. Formerly suggested as sialidase-sensitive sources of anionic groups at the cell surface of fungal species grown in chemically defined media (e.g., Fonsecaea pedrosoi), sialic acids have also been found in Sporothrix schenckii, Paracoccidioides brasiliensis, Cryptococcus neoformans and recently, in Candida albicans. Methods used involved adequate hydrolysis and extraction procedures, HPTLC, gas-chromatography, colorimetry, mass spectroscopy, lectin and influenza virus binding. Apart from protecting fungal cells against phagocytosis (S. schenckii, C. neoformans) and playing a cellular structural role (F. pedrosoi), other biological functions of sialic acids are still being investigated.