Histochemical analysis of glycoconjugates in the domestic cat testis

The localization and characterization of oligosaccharide sequences in the cat testis was investigated using 12 lectins in combination with the ßelimination reaction, N-Glycosidase F and sialidase digestion. Leydig cells expressed O-linked glycans with terminal aGalNAc (HPA reactivity) and N-glycans with terminal/internal aMan (Con A affinity). The basement membrane showed terminal Neu5Aca2,6Gal/GalNAc, Galß1,3GalNAc, a/ßGalNAc, and GlcNAc (SNA, PNA, HPA, SBA, GSA II reactivity) in O-linked oligosaccharides, terminal Galß1,4GlcNAc (RCA120 staining) and aMan in N-linked oligosaccharides; in addition, terminal Neu5Aca2,3Galß1,4GlcNac, Forssman pentasaccharide, aGal, aL-Fuc and internal GlcNAc (MAL II, DBA, GSA I-B4, UEA I, KOH-sialidase-WGA affinity) formed both O- and N-linked oligosaccharides. The Sertoli cells cytoplasm contained terminal Neu5Ac- Galß1,4GlcNAc, Neu5Ac-ßGalNAc as well as internal GlcNAc in O-linked glycans, aMan in N-linked glycoproteins and terminal Neu5Aca2,6Gal/ GalNAc in both O- and N-linked oligosaccharides. Spermatogonia exhibited cytoplasmic N-linked glycoproteins with aMan residues. The spermatocytes cytoplasm expressed terminal Neu5Aca2,3Galß1,4 GlcNAc and Galß1,3GalNAc in O-linked oligosaccharides, terminal Galß1,4GlcNAc and a/ßGalNAc in N-linked glycoconjugates. The Golgi region showed terminal Neu5aca2,3Galß1,4GlcNac, Galß1,4GlcNAc, Forssman pentasaccharide, and aGalNAc in O-linked oligosaccharides, aMan and terminal ßGal in N-linked oligosaccharides. The acrosomes of Golgi-phase spermatids expressed terminal Galß1,3GalNAc, Galß1,4GlcNAc, Forssmann pentasaccharide, a/ßGalNAc, aGal and internal GlcNAc in O-linked oligosaccharides, terminal a/ßGalNAc, aGal and terminal/internal aMan in N-linked glycoproteins. The acrosomes of cap-phase spermatids lacked internal Forssman pentasaccharide and aGal, while having increased a/ßGalNAc. The acrosomes of elongated spermatids did not show terminal Galß1,3GalNAc, displayed terminal Galß1,4GlcNAc and a/ßGalNAc in N-glycans and Neu5Ac-Galß1,3GalNAc in O-linked oligosaccharides

Histological and immunohistochemical investigation on ovarian development and plasma estradiol levels in the swordfish (Xiphias gladius 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-17b‚ (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 oogenesis

Oestrus and anoestrus changes of morphology and glycoconjugates in the apical surface of the infundibulum of mare oviducts

The oviduct plays an essential role in the mammalian reproduction and it undergoes significant endocrine-induced morphological, biochemical and physiological changes during the sexual cycle. Glycoproteins are involved in the cell-to-cell interaction as well as in the constitution of the luminal microenvironment within the oviduct. The objective of the present study was to investigate the morphological and oligosaccharide changes occurring in the apical surface of the infundibulum epithelium of mares during oestrus and anoestrus. Infundibulum fragments from two oestrus (with a follicle > 35 mm) and two anoestrus mares were processed for scanning electron microscopy (SEM) and for lectin histochemistry. SEM evidenced apical protrusions in non-ciliated cells scattered along the epithelium during the oestrus stage. Lectin binding showed a stronger labelling of luminal surface with MAL II, SNA, PNA, DBA, HPA, Con A, sialidase-WGA, GSA I-B4, GSA II and LTA during oestrus than in anaoestrus. In addition, PNA, DBA and SBA showed an increased affinity after neuraminidase digestion during oestrus. These results demonstrate a higher presence of Neu5acα2,3Galβ1,4GlcNAc, Neu5Acα2,6Gal/GalNAc, Galβ1,3GalNAc, Neu5AcGalβ1,3GalNAc, Forssman pentasaccharide, Neu5AcForssman pentasaccharide, αGalNAc, αMan, βGlcNAc, αGal and αL-Fuc during oestrus than in anoestrus. The morphological and carbohydrate changes suggest that the infundibulum activity of the mare oviduct is related to oestrogen fluctuations occurring during the sexual cycle

Espressione del recettore mu per gli oppioidi nell’epitelio seminifero di gatto

The presence of endogenous opioids has been evidenced both in the male and female reproductive tract, but less is still known on the role that they have on the reproductive function. On the contrary lots of studies evidenced the role that calcium has on capacitation and acrosome reaction, two important steps in the process that leads sperm cells to acquire the ability of fertilizing the egg. In this study by means of immunostaining techniques the mu-opioid receptor has been localized-on male cat germ cells and its presence and expression has been studied during spermatogenesis

Lectin histochemistry study in the domestic cat epididymis

The epithelium lining the mammalian epidìdymis synthesizes and secretes glycoproteins involved in maturation and Storage of sperm. This study analyzed the oligosaccharide sequence of epithelium and spermatozoa of the cat epididymis by means the lectin histochemistry. Epididymides from adult cats were fixed in Bouin's fluid and embedded in Paraplast. Sections were stained with 13 lectins in combination with sialidase (s) treatment. Basai cells show no lectinbinding sites. Apical cells reacted with s-PNA and GSA I-B4 in the cauda epididymis. Principal cells showed: 1) Golgì zone affinity to UEA I along the ductus, and no binding to MAL II, DBA, RCA120, HPA, WGA as well as no change of PNA staining after sialidase treatment in the caudal region; 2) apical zone reactivity with SBA in the whole ductus and with SNA in the cauda. Stereocilia showed a major reactivity with MAL II, PNA, s-PNA, s-DBA, RCA120, WGA in the cauda and no change in thè staining intensity with SNA, HPA, SBA, Con A along the ductus. Spermatozoa displayed a staining increase with MAL II, PNA, s-PNA, s-DBA, RCA120, HPA, SBA, WGA moving to cauda epididymis but no change with SNA and Con A. The regional differences in the lectin-binding pattern of thè epithelium and spermatozoa of cat epididymis could be related to the production of region-specific glycoproteins implicated in the maturation and/or storage of spermatozoa

Preliminary study on in vivo rooting of ornamental plants growing on peat-free growing media

Studies on the use of peat-free growing media to grow potted ornamental plants are nowadays increasing, due to environmental concerns around the exploitation of peat, but these kind of studies are lacking with respect to cutting production. In this work, we investigated rhizogenesis on cuttings of four ornamental species (Viburnum rhytidophyllum L., Pyracantha koidzumii × P. coccinea 'Mohave', Prunus laurocerasus L., Euonymus japonicus Thunb., Ligustrum sinense Lour.) planted on the following growing media: 1) peat:pumice 70:30 v v-1 (control); 2) coconut coir dust:pumice 70:30 v v-1; 3) coconut coir dust:green compost 55:45 v v-1; 4) coconut coir dust:green compost: Stabilized wood fiber 40:30:30 v v-1; 5) coconut coir dust 100 v v-1; 6) green compost 100 v v-1; 7) stabilized wood fiber 100 v v-1. Twelve cm-leafed-cuttings (with 4-6 leaves) were prepared and treated with 4000 ppm indole-3-butyric acid (IBA), 4000 ppm 1-naphthaleneacetic acid (NAA), 4000 ppm IBA+NAA or without hormones. After 120 days, rooting and shooting were evaluated considering root dry weight, length and the root area as main performance indicators. Cuttings grown on substrates with green compost and coconut coir dust generally tended to have same performances of cuttings grown on peat-based media while the cuttings grown on stabilized wood fiber media showed lower rooting grown. In general, data showed that green compost and coconut coir dust could represent excellent substitutes and alternatives to peat for the cultivation of cuttings in commercial nursery providing innovation elements regarding the total elimination of peat during the propagation phase of plants