Lectin histochemical aspects of the ovarian lamellae epithelium of Genypterus blacodes (Schneider, 1801) / Padrão de reação lectino-histoquímico do epitélio laminar ovariano em Genypterus blacodes (Schneider, 1801)

The composition and distribution of the glycoconjugates (GCs) secreted by the epithelium of ovarian lamellae with reference to the reproductive biology of Genypterus blacodes (Schneider, 1801) through lectin hi stochemistry is here discussed. In this species, the epithelial cells that line the ovarian cavity presented sharp morphological variations along the reproductive cycle related to the mucus secretion that accompanies oocyte ma turation. During sp awning season, residues of mannose and N-acetylglucosamine were detected in the glycocalyx of those cells using lectinhistochemistry. N- acetylgalactosamine and fucose were also observed in the same zone. The greatest variations in the lectinhistochemical pattern were found in the apical cytoplasm composition in comparison to the basal zone of the cells. The results of the present study were discussed by comparing their possible functional implications.

A composição e distribuição dos glicoconjugados (GCs) secretado pelo epitélio do ovário de lamelas com referência à biologia reprodutiva de Genypterus blacodes (Schneider, 1801) através da histoquímica com lectinas é aqui discutida. Nesta espécie, as células epiteliais que revestem a cavidade do ovário apresentou acentuada variação morfológica ao longo do ciclo reprodutivo relacionados com a secreção de muco que acompanha a maturação do oócito. Durante a época de desova, de resíduos de manose e N-acetilglicosamina foram detectados no glicocálix dessas células usando histoquímica de lectinas. N-acetilgalactosamina e fucose também foram observados na mesma zona. As maiores variações no padrão de lectinas foram encontradas na composição do citoplasma apical, em comparação com a zona basal das células. Os resultados do presente estudo foram discutidos, comparando as suas possíveis implicações funcionais.

Glycobiology of Leishmania donovani.

Leishmania donovani, the causative organism of visceral leishmaniasis (VL) is one of the deadliest of the entire known Leishmania species. This protozoan parasite displays immense adaptability to survive under extremely harsh conditions. Cell surface glycoconjugates play a pivotal role in parasite virulence and infectivity. This review mainly highlights on the importance of these molecules and their reported roles with special emphasis on L. donovani sialobiology. The recently evolved information reported by our group regarding the identification and characterization of sialoglycans and their possible mode(s) of acquisition as also the detailed identification, characterization of anti-O-acetylated sialic acid (anti-OAcSA) antibodies and their emerging biological roles, notably as molecules that may aid in host defense against the pathogen has been vividly discussed in this review.

Reconocimiento de residuos glicosídicos mediante lectinas conjugadas en esbozos de miembros de ave / Detection of glycoside residues in chicken limb bud using conjugate lecitines

El esbozo de miembro cuenta con un núcleo mesenquimático cubierto por un epitelio ectodérmico, los que experimentan interacciones, durante el desarrollo, conducentes a la formación de una extremidad normal. En los últimos años, utilizando la técnica de lectinas-HRP, se han estudiado la distribución y el significado de residuos glicosídicos en las interacciones epitelio-mesenquimáticas que ocurren durante la organogénesis. Así, se han identificado y caracterizado los cambios que experimentan diversos glicoconjugados en la diferenciación, reconocimiento e interacciones celulares en diversos sistemas embrionarios. Para conocer estos aspectos, hemos utilizado embriones de pollo con diferentes edades (72, 92, 120 y 168 horas), que sirvieron como dadores de los esbozos de miembros. Este material fue sometido a técnica histológica corriente y a la técnica lectinas, utilizando las siguientes lectinas-HRP:UEA (ulex europaeus agglutinin), DBA (dolichos biflorus agglutinin), ECL (erithyna cristagalli lectin), RCA (ricinus communis agglutinin), LTA (lotus tetragonolobus agglutinin), BSL (Bandeiraea simplicifolia lectin), SBA (glycine max agglutinin), además, sus respectivos inhibidores, para verificar la específidad de cada lectina. Los resultados mostraron existencia de varios residuos glicosídicos, siguiendo un patrón cambiante a lo largo, tantro a nivel del epitelio, como del mesénquima. Se analiza el siginificado de los resultados y se concluye que los patrones glicosídicos detectados peden tener algún significado en el proceso de diferenciación del esbozo en estudio.

Carbohydrate epitopes: structure and presentation and the reactivity of biological ligands: recognition of alpha-D-Galp NAc and alpha-D-Galp conformational structures

Apart from glycolipids and glycoproteins that express A and B blood group antigens which contain terminal nonreducing units of alpha-D-Galp NAc and alpha-D-Galp respectively, there are several other glycoconjugates in nature that contain these units linked to unfucosylated saccharides or protein. They represent normal products of the action of specific glycosyl-transferases in primate and nonprimate mammalian cells, protozoa and a few other microorganisms, end-units of carbohydrate components that have not benn further processed by additional glycosylation, or neo-antigens resulting from deregulation of certain transferases as in tumor cells. Biological ligands recognizing these structures include mono and polyclonal antibodies, bacterial fimbriae and laminin. Binding depends on the linkages and sequence of the carbohydrate chain, but also on the epitope conformation as influenced by adjacent substitution, angling determined by the glycoconjugate-substrate interaction, steric hindrance and other factors. These aspects are discussed in this minireview.

Strategies for the structure determination of parasite glycoconjugates using fast atom bombardment mass spectrometry

Fast atom bombardment mass spectrometry (FAB MS) is a sensitive and powerful technique for the analysis of thermally labile and involatile materials. In this paper we describe the application of the method to the characterization of oligosaccharides isolated from the cell surface glycoconjugates of parasitic protozoa. The spectra typically contain both molecular ions, which define the monosaccharide composition, and fragment ions which are related to the structure of the intact molecules. The use of additional techniques such as chemical derivatization and colisional activation enhances fragmentation and simplifies interpretation of the data, enabling the determination of residue sequence, the positions of branch points, and the location of noncarbohydrate substituents. We have applied these techniques to the characterization of phosphoinositol oligosaccharides from members of the Trypanosoma family, including Leptomonas smueli, Endotrypanum schaudinni and Leishmania adleri. Although it is not usually possible to determine the complete structure of oligosaccharide by mass spectrometric methods alone, the information gained greatly simplifies the interpretation of the results from other techiniques such as nuclear magnetic resonance and methylation analysis.

Structural features of LPPG and related compounds isolated from Trypanosoma cruzi trypomastigotes

A hydrophobic fraction isolated from trypomastigotes of Trypanosoma cruzi is being characterized using immunological and chemical techniques. The lipopeptidophosphoglycan (LPPG) was identified in this fraction since it gave a positive reaction with anti-LPPG rabbit serum and had similar structural features such as the presence of ceramide as the lipid moiety, furanoic galactose, and a glycan moiety consistent with that obtained from an authentic sample of epimastigote LPPG, as judged by thin-layer chromatography. Furthermore, the hydrophobic fraction contained other glycolipids with different structural features. The lipid moiety of these compounds is alkylglycerol rather than a ceramide, the carbohydrate chain appears to be less complex than that in LPPG and no reactivity was observed towards an anti-LPPG serum

Evidence for the presence of ß-D-galactofuranosyl residues (1->3)-linked to a-D-mannopyranose on the cell membrane in all developmental stages of Trypanosoma cruzi

The lipopeptidophosphoglycan (LPPG) component isolated from epimastigote forms of Trypanosoma cruzi has antigenic properties. Using a rabbit antiserum aginst LPPG, specific for ß-D-galactofuranosyl residues (1-3)-linked to alfa-Dmannopyranose, we identified, by indirect immunofluorescence, the presence of epitopes containing ß-D-galactofuranosyl structures on the cell membrane of epimastigote, amastigote and trypomastigote forms of the parasite. The results demonstrade that all developmental stages of T. cruzi contain similar antigenic determinants on their cell body and flagellar membrane