Glycopattern analysis and structure of the egg extra-cellular matrix in the Apennine yellow-bellied toad, Bombina pachypus (Anura: Bombinatoridae)

We studied the glycopatterns and ultrastructure of the extra-cellular matrix (ECM) of the egg of theApennine yellow-bellied toad Bombina pachypus, by light and electron microscopy in order to determine structure,chemical composition and function. Histochemical techniques in light microscopy included PAS and AlcianBlue pH 2.5 and 1.0, performed also after b-elimination. Lectin-binding was tested with nine lectins (AAA,ConA, DBA, HPA, LTA, PNA, SBA, UEA-I, WGA). An inner fertilization envelope (FE) and five jelly layers(J1–J5) were observed, differing in histochemical staining, lectin binding and ultrastructure. Most glycans wereO-linked, with many glucosamylated and fucosylated residues. The fertilization envelope presented a perivitellinespace and a fertilization layer, with mostly neutral glycans. The jelly layers consisted of fibers and granules,whose number and orientation differed between layers. Fibers were densely packed in J1 and J4 layers,whereas a looser arrangement was observed in the other layers. Jelly-layer glycans were mostly acidic and particularlyabundant in the J1 and J4 layers. In the J1, J2 and J5 layers, neutral, N-linked glycans were also observed.Mannosylated and/or glucosylated as well as galactosyl/galactosaminylated residues were more abundant in theouter layers. Many microorganisms were observed in the J5 layer. We believe that, apart from their functions inthe fertilization process, acidic and fucosylated glycans could act as a barrier against pathogen penetration

A histochemical approach to glycan diversity in the urothelium of pig urinary bladder

Intracellular glycans in the urothelium of urinary bladder of 10 adult male Landrace pigs were characterized in situ by immunohistochemical detection of Muc1 mucin by anti MUC1 from rabbit, conventional histochemical techniques (Periodic-Acid Schiff, Alcian Blue pH 2.5, High-Iron Diamine), and binding with 13 lectins (PNA, DBA, RCA-I, WGA, SBA, BSI-B4, ConA, AAA, UEA-I, LTA, LFA, MAA-II, SNA) combined with chemical and enzymatic pre-treatments (β-elimination, desulfation and neuraminidase) to gather reference data for this model animal. Muc1 mucin was detected in the secreting granules of superficial cells and the underlying layer of intermediate cells. The secreting granules in both intermediate cells and superficial cells were rich in carbohydrates, with the oligosaccharidic chains mostly O-linked to proteins. Glycoproteins were prevailing over glycosaminoglycans (GAGs). In both superficial and intermediate cells sulfated and/or sialylated glycans were present, sulfation decreasing in the deeper layers. Lectin-binding detected presence of terminal sialic acid linked mostly in α2,6 to GalNAc, Gal terminal or subterminal to sulfates, GalNAc, GlcNAc, and Fuc, mostly linked in α1,6, α1,3 α1,4 and α1,2 to GlcNAc or Gal, but not to lactosamine chains. Except for fucosylation, the oligosaccharidic chains in the glycoproteins of the urothelium of pig urinary bladder were similar to those linked to human MUC1, which is fundamental in cell adhesion and immunological processes in the urothelium. The co-distribution of Muc1 and saccharidic residues suggests that many of them are linked to the glycoprotein

Histochemical investigations on the secretory cells in the oesophagogastric tract of the Eurasian green toad, Bufo viridis

Summary The secretory cells of the oesophagogastric tract of the Eurasian toad, Bufo viridis, were examined using standard histochemical methods and lectin histochemistry. Two goblet cell types were found in the oesophageal epithelium, differing in their morphology and the histochemical features of the secretory granules. These contained mainly acidic glycoconjugates, both sulphated and carboxylated, and a small amount of pepsinogen. Type I goblet cells contained stable class-III mucosubstances, which were absent in Type II. No pluricellular oesophageal glands were found. The oesophagogastric junction had a superficial epithelium similar to that of the oesophageal epithelium, with alveolar pluricellular glands, secreting stable class-III mucins, and few oxynticopeptic cells. The gastric mucosa presented secretory cells both in the surface epithelium and in the gastric glands. Superficial and foveolar cells produced neutral mucins with Galβ1,3GalNAc residues. Neck cells, oxynticopeptic cells and endocrine cells were found in the gastric glands. Neck cells produced stable class-III mucosubstances. A functional gradient was observed in the oxynticopeptic cells from the oral to the aboral fundus, with a decrease in pepsinogen secretion towards the aboral fundus and a possible increase in HCl secretion. In the pyloric mucosa, the oxynticopeptic cells disappeared and the glands produced only neutral mucins, without stable class-III mucosubstances

The phylogenetic relationships of Triturus italicus

Several studies attempted to reconstruct the phylogenetic relationships among the species of Tritums by a number of data sets from morphology, courtship behaviour, reproductive interactions, cytogenetics and molecules. The present paper critically reviews those studies with regard to the phylogenetic affinities of T. italicus (Peracca, 1898). Most studies locate T. italicus within the radiation of the small-sized newts (subgenus Palaeotriton), but are in conflict about its sister-group, whether T. vulgaris or T. boscai. Recent studies on mtDNA sequences suggest that Triturus is paraphyletic and several other genera should be included within the genus, with T. italicus being an outgroup of all the other species and genera. It is concluded that, at present, little can be stated about the relationship of T. italicus and that new data should be collected