Glycocomposition of the apocrine interdigital gland secretions in the fallow deer (Dama dama)

The secretions of the tubular interdigital glands were investigated by conventional (Periodic-Acid Schiff, Alcian-Blue at different pH, Low Iron Diamine and High Iron Diamine) and lectin (Con-A, UEA-I, LTA, WGA, GSA-II, GSA-IB4, SBA, PNA, ECA, DBA, MAL-II and SNA) histochemical methods in adult males and females of different age of fallow deer during the breeding season. Sialidase digestion and deacetylation pre-treatment were also employed in conjunction with lectin histochemistry. The glandular epithelium consisted of a single layer of low columnar cells with typical apical protrusions. No substantial differences of the above histochemical staining in relation to sex and age were observed. Conventional histochemical staining revealed that the interdigital glands secreted neutral glycoproteins whereas acidic glycocomponents did not seem to be present. Lectin histochemical technique allowed us to disclose a great heterogeneity of glycoproteins with N- and O-linked oligosaccharides containing alpha-D-Man/alpha-D-Glc, GlcNAc, alpha-Fuc, terminal beta-D-Gal-(1-3)-D-GalNAc, -D-Gal-(1-4)-D-GlcNAc, alpha-Gal and beta-GalNAc residues. beta-GalNAc and disaccharide beta-D-Gal-(1-3)-D-GalNAc were also found as subterminal to sialyl moieties. The lack of sexual and age-related differences in the glucidic content of the glandular secretions seems to indicate that the glycoderivatives may play only an accessory role in the production of odoriferous signals in fallow deer

Glycohistochemical investigation on canine and feline zonae pellucidae of preantral and antral oocytes

Glycoconjugate modifications were analysed in the zona pellucida during development of oocytes in dog and cat using conventional histochemical staining methods with or without previous carbohydrate digestion. A series of lectins combined with desulphation and sialic acid degradation were applied. No differences were observed between dog and cat follicles using conventional histochemical staining methods. In both species, the zona pellucida and follicular fluid/intercellular matrix strongly reacted with PAS and high iron diamine stain (HID) and reacted moderately with low iron diamine stain (LID). Treatment with testicular hyaluronidase, chondroitinase ABC, chondroitinase AC and chondroitinase B treatment diminished HID and LID positivity of follicular fluid and intercellular matrix. Lectins that gave the most intense staining of the zona pellucida of both species were SBA, PNA, RCA-I, GSA-IB4 and WGA, indicating the presence of beta-D-GalNAc, D-Gal and GlcNAc residues. Sulpho- and asulpho-carbohydrates were identified in terminal and/or subterminal positions linked to sialic acid residues. In conclusion, the results indicate that glycosaminoglycans are not present in the zona pellucida of both species. Differences were observed in carbohydrate residues and in their spatial distribution, depending on species and developmental stage of the follicles. The similarity in lectin affinity between ooplasm and zona pellucida of oocytes present in follicles at different stages of development confirm the involvement of oocytes in zona pellucida production

Glycoconjugates in small antral ovarian follicles of the river buffalo (Bubalus bubalis L.)

Types and distribution patterns of glycoconjugates in antral ovarian follicles were investigated in the buffalo, using periodic-acid Schiff (PAS), high iron diamine (HID), low ion diamine (LID) and lectin histochemical staining methods. HID and LID staining procedures were preceded in some cases by digestion with testicular hyaluronidase, Streptomyces hyaluronidase, chondroitinase ABC and heparitinase (heparinase III). Lectin staining was performed with the use of 12 horseradish peroxidase (HRP) lectin conjugates. Some lectin staining procedures were preceded by neuraminidase digestion and saponification. Large amounts of isomeric chondroitin sulphates and a minor quantity of heparan sulphate and hyaluronic acid and/or chondroitin were found in follicular fluid. Lectin staining of buffalo follicular fluid revealed glycoconjugates with different glucidic determinants such as beta-N-acetylgalactosamine, beta-galactose-(1-3)-N-acetylgalactosamine, beta-galactose-(1-4)-N-acetylglucosamine, N-acetylglucosamine, alpha-fucose and alpha-glucose/alpha-mannose, and sialic acid residues. Glycosaminoglycans were absent in the zona pellucida of oocytes in small antral follicles. Acidic glycoconjugates in the zona pellucida were caused by sulphated groups and sialic acid residues. Our data show few internal glucidic residues, such as N-acetylglucosamine in the buffalo zona pellucida but many subterminal beta-N-acetylgalactosamine, alpha- and beta-galactose determinants masked by sialic acids. These findings demonstrate that buffalo follicular fluid has a very heterogeneous composition that is similar to that found in small and large bovine follicles. No differences in composition of the follicular fluid were observed in the follicles examined

Prisutnost imunoreaktivnih ciklooksigenaza u sjemenskim kanalićima i nuzjajima u alpaka prije spolne zrelosti i odraslih alpaka (Lama pacos).

In this study, we report the cell specific expression of cyclooxygenase (COX) enzymes, COX1 and COX2, in the ductuli efferentes and epididymis of prepubertal and adult alpaca. COX1 weakly stained the cytoplasm of epithelial cells lining the ductuli efferentes and the epididymal corpus, whereas these cells were immunonegative in the epididymal caput and cauda. In adults, we observed an increase in the immunsignals for COX1 in the cytoplasm of epithelial cells lining the ductuli efferentes and all the epididymal regions. In prepubertal alpaca, immunoreactivity for COX2 was not revealed in the epithelial cells lining the ductuli efferentes and epididymal regions, whereas it was evidenced in adult animals. The apical rich mithocondria cells immunoreacted only with COX1 in the epididymis of prepubertal animals, whereas they expressed both COX1 and -2 in the adult alpaca. Our results suggest that COXs may play a role in the pubertal development of the excurrent duct system of the alpaca.Istražena je stanično specifična ekspresija enzima ciklooksigenaza COX1 i COX2 u sjemenim kanalićima i nuzjaju u alpaka prije spolne zrelosti i odraslih alpaka. Slaba aktivnost COX1 dokazana je u citoplazmi epitelnih stanica sjemenih kanalića i tijela nuzjaja dok su iste stanice bile imunološki negativne u glavi i repu nuzjaja. U odraslih je opaženo povećanje imunosnog signala za COX1 u citoplazmi bazalnih epitelnih stanica sjemenih kanalića i cijelog područja nuzjaja. U alpaka prije spolne zrelosti, imunoreaktivnost za COX2 se nije vidjela u epitelnim stanicama sjemenih kanalića i nuzjajima kao što je to bilo kod odraslih životinja. Apikalne stanice bogate mitohondrijima reagirale su samo na COX1 u nuzjajima životinja prije spolne zrelosti dok je ekspresija i COX1 i COX2 opažene u odraslih alpaka. Rezultati ukazuju na moguću ulogu ciklooksigenaza u razvoju razgranatog sustava kanalića prije spolne zrelosti u alpaka

Variations in the lectin-binding on the zona pellucida during oocyte growth in some wild ungulates

The aim of the present study was to examine the glycoconjugate modifications occurring in the zona pellucida during oocyte growth in fallow, red and roe deer using a battery of lectins combined with sialidase digestion and chemical treatments. This histochemical approach allowed us to sequence the oligosaccharidic side chains of the zona pellucida glycoproteins in these wild ungulates. The most effective lectins in the zona pellucida of these species were SBA, PNA, RCA-I GSA-IB4, and WGA, indicating the presence of beta-D-N-Acetylgalactosamine, beta-D-Galactose, alpha-D-Galactose and N-Acetylglucosamine residues. Additionally, sialic acid moieties were demonstrated. We also observed differences in the glycosidic residue content and in their spatial distribution, depending on the species and stage of follicle development

Histochemical detection of the lectin-binding carbohydrates in the zona pellucida during oocyte growth in the wild boar (Sus scrofa scrofa)

The changes that occur in the carbohydrate composition of zona pellucida glycoproteins during oocyte maturation in the wild-boar were studied using periodic-acid Schiff (PAS), High Iron Diamine (HID) and Low Iron Diamine (LID). Lectin staining was performed with a panel of 11 HRP-lectin conjugates combined with neuraminidase digestion and chemical treatments. There were few internal glucidic residues, such as N-acetylglucosamine, in the wild boar zona pellucida but there were many subterminal beta-N-acetylgalactosamine, alpha- and beta-galactose determinants masked by sialic acid. In addition, beta-N-acetylgalactosamine, beta-galactose-(1-3)-N-acetylgalactosamine and beta-galactose-(1-4)-N-acetylglucosamine were detected in the sulphated form in the terminal and/or subterminal position. Some differences in the lectin reactive sites occurred in the zona pellucida, depending on the stage of oocyte maturatio

Prisutnost imunoreaktivnih ciklooksigenaza u sjemenskim kanalićima i nuzjajima u alpaka prije spolne zrelosti i odraslih alpaka (Lama pacos).

In this study, we report the cell specific expression of cyclooxygenase (COX) enzymes, COX1 and COX2, in the ductuli efferentes and epididymis of prepubertal and adult alpaca. COX1 weakly stained the cytoplasm of epithelial cells lining the ductuli efferentes and the epididymal corpus, whereas these cells were immunonegative in the epididymal caput and cauda. In adults, we observed an increase in the immunsignals for COX1 in the cytoplasm of epithelial cells lining the ductuli efferentes and all the epididymal regions. In prepubertal alpaca, immunoreactivity for COX2 was not revealed in the epithelial cells lining the ductuli efferentes and epididymal regions, whereas it was evidenced in adult animals. The apical rich mithocondria cells immunoreacted only with COX1 in the epididymis of prepubertal animals, whereas they expressed both COX1 and -2 in the adult alpaca. Our results suggest that COXs may play a role in the pubertal development of the excurrent duct system of the alpaca.Istražena je stanično specifična ekspresija enzima ciklooksigenaza COX1 i COX2 u sjemenim kanalićima i nuzjaju u alpaka prije spolne zrelosti i odraslih alpaka. Slaba aktivnost COX1 dokazana je u citoplazmi epitelnih stanica sjemenih kanalića i tijela nuzjaja dok su iste stanice bile imunološki negativne u glavi i repu nuzjaja. U odraslih je opaženo povećanje imunosnog signala za COX1 u citoplazmi bazalnih epitelnih stanica sjemenih kanalića i cijelog područja nuzjaja. U alpaka prije spolne zrelosti, imunoreaktivnost za COX2 se nije vidjela u epitelnim stanicama sjemenih kanalića i nuzjajima kao što je to bilo kod odraslih životinja. Apikalne stanice bogate mitohondrijima reagirale su samo na COX1 u nuzjajima životinja prije spolne zrelosti dok je ekspresija i COX1 i COX2 opažene u odraslih alpaka. Rezultati ukazuju na moguću ulogu ciklooksigenaza u razvoju razgranatog sustava kanalića prije spolne zrelosti u alpaka

Immunopresence and functional activity of prostaglandin-endoperoxide synthases and nitric oxide synthases in bovine corpora lutea during diestrus.

The aim of this study was to evaluate the occurrence and the activity of prostaglandin- endoperoxide synthase 1 (PTGS1), PTGS2, and endothelial, neuronal, and inducible nitric oxide synthase (e-, n-, and iNOS) in early, mid, late, and regressive corpora lutea (CL) of bovines during diestrus. PTGS1 immunoreactivity was localised mainly in the cytoplasm of small luteal cells, whereas PTGS2 was detected in the cytoplasm of large luteal cells during early, mid, and late stages. The immunoexpression of all NOS isoforms was observed in the nuclei of luteal cells in the CL stages examined. PTGS1 enzyme activity was higher in late CL and lower in regressive ones; PTGS2 increased from early to late CL and lowered in regressive ones. Constitutive NOS enzymatic activity (eNOS plus nNOS) was higher in late CL and lower in regressive ones; iNOS was lower in regressive CL. These results support the idea that PTGSs and NOSs regulate the bovine CL life span mainly during the transition from the luteotrophic to the luteolytic phase

Apoptotic cell death in canine hair follicle

Apoptotic cell death is an essential homeostatic mechanism involved in the control of cellular turnover in a variety of adult tissues. Cytoplasmic and nuclear condensation morphologically define this process whose biochemical hallmark is extensive DNA fragmentation into discrete oligonucleosomic units. Hair follicle growth and regression has been shown to be correlated with apoptosis in humans, mice, rats and guinea pigs. The present study was carried out to evaluate its implication in canine hair biology in order to define the spatio-temporal relationship between apoptosis and the hair cycle in dogs. As assessed by terminal deoxy-nucleotidyl transferase-mediated d-UTP nick-end-labelling (TUNEL) and by basic histological and ultrastructural assays, apoptotic cells appeared both in the growing and in the regressing follicle epithelium showing the well characterized morphological features described in the previous relevant literature