Differential lectin binding patterns in the oviductal ampulla of the horse during oestrus

We investigated the oligosaccharide sequence of glycoconjugates, mainly sialoglycoconjugates, in the horse oviductal ampulla during oestrus by means of lectin and pre-lectin methods such as the KOH-neuraminidase procedure to remove sialic acid residues and incubation with N-glycosidase F to cleave N-linked glycans. Ciliated cells displayed N-linked oligosaccharides throughout the cytoplasm. The cilia glycocalyx expressed both N- and O-linked (mucin-type) oligosaccharides, both showing a high variety of terminal sequences. In the most non-ciliated cells, the whole cytoplasm contained N-linked oligosaccharides with terminal αGal as well as mucin-type glycans with terminal Forssman pentasaccharides. In a few scattered non-ciliated cells, the whole cytoplasm displayed sialylated N-linked oligosaccharides with terminal Neu5Ac-GalNAc and O-linked glycans terminating with neutral and/or αGalNAc, Neu5Acα2,6Gal/ GalNAc, Neu5AcGalβ1,3GalNAc. Supra-nuclear granules, probably Golgi zones, of non-ciliated cells showed mainly O-linked glycans rich in sialic acid residues. The luminal surface of non-ciliated cells showed N-linked oligosaccharides, containing terminal/internal αMan/αGlc, βGlcNAc and terminal αGal, as well as mucin-type oligosaccharides terminating with a large variety of either neutral saccharides or sialylated sequences. Apical protrusions containing O-linked oligosaccharides with terminal Forssman pentasaccharide, Neu5Ac-Galβ1,4GlcNAc, Neu5Ac-GalNAc were seen in nonciliated cells scattered along the epithelium. These findings show the presence of sialoglycoconjugates in the oviductal ampulla epithelium of the mare and the existence of different lectin binding profiles between ciliated and non-ciliated (secretory) cells, as well as the presence of non-ciliated cell sub-types which might determine functional differences along the ampullary epithelium of mare oviduct

Distribution of sialoglycoconjugates in the oviductal isthmus of the horse during anoestrus, oestrus and pregnancy: a lectin histochemistry study

The distribution of sialic acid residues as well as other glycosidic sugars has been investigated in the horse oviductal isthmus during anoestrus, oestrus and pregnancy by means of lectin and pre-lectin methods. Ciliated cells and non-ciliated (secretory) cells exhibited different lectin binding profiles that were found to change during the investigated stages. Ciliated cells did not show any reactivity in the basal cytoplasm, while the supra-nuclear cytoplasm displayed a few of oligosaccharides with terminal and internal amannose (Man) and/or aglucose (Glc) during oestrus and pregnancy and a moderate presence of oligosaccharides terminating in afucose (Fuc) during oestrus; cilia exhibited a more complex glycoconjugate pattern for the presence of oligosaccharides terminating in N-acetylgalactosamine (GalNAc), GalNAca1,3 GalNAca1,3galactose(Gal)b1,4Galb1,4N-acetylglucosamine( GlcNAc), Fuc, sialic acid (Neu5Ac)-aGalNAc belonging or not to the GalNAca1,3GalNAca1,3 Galb1,4 Galb1, 4GlcNAc sequence, and.aGalNAc and Neu5Aca 2,6Gal/GalNAc increased during oestrus. Cilia displayed terminal Galb1,3 GalNAc in pregnancy, terminal aGal in anoestrus and pregnancy and terminal or internal D-GlcNAc during anoestrus and pregnancy, respectively. The whole cytoplasm of non-ciliated cells showed oligosaccharides terminating with aGalNAc, Neu5Aca2,6Gal/GalNAc, Neu5Ac GalNAca 1,3GalNAca1,3Galb1,4Galb1,4GlcNAc during the investigated stages, as well as GlcNAc in anoestrus and pregnancy. The supra-nuclear zone of non-ciliated cells exhibited oligosaccharides with terminal Galb1,4GlcNAc and internal Man during oestrus and pregnancy as well as terminal aGal and Fuc in oestrus and Neu5Ac-Galb1,3GalNAc in pregnancy. The luminal surface of non-ciliated cells showed glycans terminating with aGalNAc and/or Neu5Ac GalNAca1,3 GalNAca1,3Galb1,4Galb1,4GlcNAc in all specimens, oligosaccharides with terminal Galb1,4GlcNAc and internal Man during oestrus and pregnancy, Neu5Ac a2,6Gal/GalNAc in anoestrus and oestrus, and glycans terminating with Galb1,3GalNAc, Neu5A aca2,3 Galb1, 4GlcNac, Neu5ac- Galb1,3GalNAc, Neu5Ac-Galb1,4 GlcNAc in pregnancy. These findings show the presence of sialoglycoconjugates in the oviductal isthmus of the mare as well as the existence of great modifications in the glycoconjugates linked to different physiological conditions

Histological and immunohistochemical investigation on ovarian development and plasma estradiol levels in the swordfish (<i>Xiphias gladius</i> L.)

The paper reports a histological and immunohistochemical description of oocyte growth and ultrastructural aspects of zona radiata (ZR) formation as well as the relationship between plasma estradiol-17&#x03B2;, (E2 ) levels and ovarian development in swordfish (Xiphias gladius L.) from the Mediterranean Sea. Ovaries were inactive during March to mid April; maturation occurred during late April to June and spawning in June and July. Zona radiata formation starts, as Pas positive material, in oocytes at the lipid stage. In this stage a deposit of electrondense material between oolemma and follicular cells appears. In the cortical alveoli stage and through the early vitellogenic stage, the deposition of a moderately electrondense material occurred on the inner side of the ZR. Finally, in late vitellogenic oocytes a third layer, made of microfibrillar material, appeared. The immunohistochemical analyses revealed that the initial internalisation of hepatic zona radiata proteins (Zrp) in the swordfish oocyte starts before the uptake of vitellogenin (Vtg) and that it is associated with the low previtellogenic E2 plasma levels, while a significant E2 increase in plasma is associated with the beginning of Vtg uptake. This would appear to confirm the hypothesis that the differential and sequential induction of zonagenesis and vitellogenesis may reflect a general feature of teleost oogenesi

Lectin histochemistry on the dorsal epidermis of the Breton dog

Expression of sugar residues and the nature of oligosaccharide linkage during keratinocyte maturation in the epidermis of the Breton dog were studied with the use of lectin histochemistry. Thirteen lectins were used. Labelling was not observed with GSA I-B4, GSA II, UEA-I, and LTA. The cytoplasm of keratinocytes reacted with PNA, HPA, Con A, and WGA from the basal layer to the granular layer. PNA and Con A showed highest reactivity in the granular cell layer. The cell surface showed increased reactivity with PNA, HPA, and WGA with maturation of keratinocytes. KOH-neuraminidase treatment (KOH-Neu) increased PNA and RCA120 staining during keratinocyte differentiation thus indicating an increase in oligosaccharides terminating with sialic acid-Galβ(1,3)GalNAc and sialic acid-Galβ(1,4)GlcNAc, respectively. Labelling of the glycocalyx of basal and spinous keratinocytes with SNA and MAA revealed terminal Neu5acα(2,6)Gal/GalNAc and Neu5acα(2,3)Galβ(1,4)GlcNAc. KOH-Neu-DBA showed oligosaccharides terminating with sialic acid-GalNAcα(1,3)GalNAc in the spinous and granular layers. A selective glycocalyx labelling of granular keratinocytes was observed with DBA and SBA. Reactions with MAA, PNA, DBA, RCA120, SBA, HPA, and WGA disappeared after the b-elimination reaction. Our findings indicate that Breton dog epidermis contains more O-linked than N-linked oligosaccharides and confirm that different subpopulations of keratinocytes can be distinguished by lectin histochemistry

Lectin histochemistry on the apocrine sweat glands of dog dorsal skin

The secretory segment of apocrine sweat glands of the dog dorsal skin was studied by means of both conventional carbohydrate histochemistry and lectin histochemistry. Conventional glycoconjugate histochemistry revealed mostly neutral glycoproteins and partly acidic non sulphated glycoproteins in the apical protrusions of the secretory cells as well as in the luminal secretions of the glands. The lectin histochemistry showed Con A binding sites in the whole cytoplasm of the secretory cells. The supra-nuclear granules (probably including also Golgi elements) were labelled by SNA, MAA, RCA120, Con A, and GSA I-B4 thus indicating the presence of sialylated or asialylated N-linked oligosaccharides. The luminal surface of secretory cells reacted with all the lectins employed except with SBA. The apocrine protrusions revealed a general decrease in the lectin staining intensity, and lacking of GSA I-B4, UEA-I, and LTA reactivity. The luminal homogeneous matrix labelled SNA, WGA, GSA II, and GSA I-B4