Ultrastructure and secretory nature of the seminal glomus in budgerigars (Melopsittacus undulatus)

The seminal glomera from captive budgerigars were dissected and prepared for ultrastructural and histochemical studies of the lining epithelia. The general structure suggested that they are formed by convolutions of the terminal portions of the ductus deferens which appear as a network of tubules. The epithelium lining the tubules was identified as pseudostratified ciliated and non-ciliated columnar epithelium. With the electron microscope it was possible to identify two different cell types in the epithelium: type 1, ciliated cells and type 2, non-ciliated epithelial cells. Light microscopy revealed periodic acid Schiff (PAS) positive material, resistant to diastase digestion in the distal parts of some of the epithelial cells, indicating its glycoprotein nature. Alcian blue/PAS staining showed mixed acidic and neutral glycoproteins. Alcian blue staining at different hydrogen ion concentrations (pH) showed that the acidic glycoprotein was of the weakly sulphated type. Periodic acid thiocarbohydrazide silver proteinase staining, at the ultrastructural level, confirmed the presence of an intracellular glycoprotein.     

Histological and histochemical characterization of the mucosa of the digestive tract in flower fish (Pseudophoxinus antalyae)

The wall of the digestive tract is composed of the tunica mucosa, tunica muscularis and tunica serosa. Lamina-submucosa separation or any glands were not observed in tunica mucosa. Goblet cells were determined to constitute a much larger reserve at digestive tract mucosa. Histochemical analysis of the intestine of flower fish (Pseudophoxinus antalyae) showed that gastrointestinal mucous content included sulphate-esters and/or carboxylic [Alcian blue (AB) 0.06+], glycogene and/or oxidable dioles [periodic acid/Schiff+ (PAS+)], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS) and strong acid sulphated [Aldehyde fuchsin+ (AF+)] glycoproteins (GPs). Except these mucosubstances to lower densities, densely sulphate (AB pH 2.5+), O-sulphate esters (AB pH 1+) strong and weak sulphated (AB 0.3 M+), GPs were also determined.     

Harderian Gland in Canadian Ostrich (Struthio camelus): A Morphological and Histochemical Study

Heads of ten healthy adult ostrich obtained from slaughter house were the constituted materials of the study. The Harderian gland (HG) was dissected out, and all of the gross morphometrical parameters including length, width and thickness as well as weight of left and right glands were recorded. Tissue sections were stained, using haematoxylin and eosin, Masson trichrome, periodic acid‐Schiff and Alcian blue (pH 2.5) techniques. In ostrich, HG was an orbital organ located ventromedially around the posterior part of the eyeball. It was an oval flatted shape, light pink colour with irregular outline and was pointed in the dorsal end. Its mean length was 35.30 ± 2.84 mm and 35.55 ± 3.58 mm in left and right sides, respectively, and mean width 15.30 ± 1.20 mm and 15.65 ± 1.18 mm in left and right sides, respectively. There was no significant difference between length, thickness, weight and width of left and right glands. Histological results showed that the glandular epithelium was multilobular and compound tubuloalveolar. The gland was surrounded by a connective tissue capsule, and the epithelium was lined by simple columnar epithelial cells of varying height. The secretion of HG was mucous and the secretion type was apocrine. Mucosubstance analysis revealed that secretory units contained acidic and neutral glycoproteins. The granules within the epithelial cells lining the intralobular and inter‐lobular excretory ducts of the gland were positive for periodic acid‐Schiff and Alcian blue (pH 2.5).     

Morphofunctional description of mucous cells in the gills of the Arapaimidae Arapaima gigas (Cuvier) during its development

The gill structure of the Amazonian fish Arapaima gigas (Cuvier 1829) shows ontogenetic changes during development, particularly due the transition from the aquatic to the obligatory air breathing mode of respiration. However, three main cell types can be found in the gills: mitochondrial rich cells, pavement cells and mucous cells (MCs). The MCs are involved in the secretory pathway. The functions of the secreted molecules include mechanical protection of epithelia, protection against parasites and bacterial infection, and role on ion regulation. In this study, we analysed mucous cell location and mucous cell type, based on pH, during the development of A. gigas. Using samples obtained from the environment, gills were collected and fixed in buffered solution. Histological techniques for the identification of MCs were performed Alcian Blue (AB) and periodic acid‐Schiff (PAS). The results showed the presence of PAS+ and AB+ cells in the whole filament in all examined fish. In animals less than 50 g, few MCs were present, and no differences were observed in AB+ and PAS+ cells. In animals weighing close to 500 g, more PAS+ cells than AB+ cells were observed, and in animals that weighed more than 1,000 g, more AB+ cells than PAS+ cells were observed. These observations may be a result of the ontogenetic changes in the gill epithelia, which can change the osmorespiratory compromise in ion regulation functions as well the glycosaminoglycans secreted by PAS cells, which in large animals can play a role in the protection against parasites and bacterial infection.     

Histochemical Analysis of Glycoconjugates in the Skin of a Catfish (Arius Tenuispinis, Day)

A histochemical study using conventional carbohydrate histochemistry (periodic‐acid staining including diastase controls, alcian blue staining at pH 1 and 2.5) as well as using a battery of 14 fluorescein isothiocyanate (FITC)‐labelled lectins to identify glycoconjugates present in 10 different areas of the skin of a catfish (Arius tenuispinis) was carried out. The lectins used were: mannose‐binding lectins (Con A, LCA and PSA), galactose‐binding lectins (PNA, RCA), N‐acetylgalactosamine‐binding lectins (DBA, SBA, SJA and GSL I), N‐acetylglucosamine‐binding lectins (WGA and WGAs), fucose‐binding lectins (UEA) and lectins which bind to complex carbohydrate configurations (PHA E, PHA L). Conventional glycoconjugate staining (PAS staining, alcian blue at pH 1 and 2.5) showed that the mucous goblet cells contain a considerable amount of glycoconjugates in all locations of the skin, whereas the other unicellular gland type, the club cells, lacked these glycoconjugates. The glycoproteins found in goblet cells are neutral and therefore stain magenta when subjected to PAS staining. Alcian blue staining indicating acid glycoproteins was distinctly positive at pH 1, but gave only a comparable staining at pH 2.5. The mucus of the goblet cells therefore also contains acid glycoproteins rich in sulphate groups. Using FITC‐labelled lectins, the carbohydrate composition of the glycoproteins of goblet cells could be more fully characterized. A distinct staining of the mucus of goblet cells was found with the mannose‐binding lectins LCA and PSA; the galactosamine‐binding lectins DBA, SBA and GLS I; the glucosamine‐binding lectin WGA; and PHA E which stains glycoproteins with complex carbohydrate configurations. No reaction occurred with the fucose‐binding lectin UEA and the sialic acid‐specific lectin SNA. In addition, the galactose‐binding lectins PNA and RCA showed only a weak or completely negative staining of the mucus in the goblet cells. The specificity of the lectin staining could be proved by inhibiting binding of the lectins by competitive inhibition with the corresponding sugars. From these data, we can conclude that the mucus produced by the epidermal goblet cells of A. tenuispinis is rich in mannose, N‐acetylgalactosamine and N‐acetylglucosamine residues.     

Lectin binding pattern of gastric mucosa of pacific white-sided dolphin, Lagenorhynchus obliquidens

The stomach of the Pacific white-sided dolphin is divided into three parts: forestomach, proper gastric gland portion, and pyloric chamber. The histological features of the dolphin stomach are similar to those of terrestrial mammal stomachs, although the distribution of glycoconjugates in mucosal cells of the dolphin stomach is unknown. To learn about glycoconjugates in cetacean gastric mucosa, the glycoconjugate distribution in the mucous epithelium of the Pacific white-sided dolphin was studied using 21 lectins. Among the lectins tested, GSL-I and DBA specifically labelled the superficial layer of the forestomach epithelium. GSL-I, SBA, RCA-I, VVA, GSL-II, DSL, LEL, STL, s-WGA, WGA, PNA, and Jacalin labelled the luminal surface of the chief cells in the proper gastric gland. GSL-I, SBA, RCA-I, DSL, LEL, STL, s-WGA, PNA, and LCA labelled tubular structures in the cytoplasm of parietal cells. The surface portion of the pits in the pyloric chamber strongly reacted with RCA-I, GSL-II, WGA, PNA, LCA, PHA-L, and UEA-I, whereas the neck portion reacted weakly. Although lining one tubular portion, individual secretory cells in the pyloric gland displayed a heterogeneous reaction. This is the first report on the lectin histochemistry of a cetacean stomach and reveals GSL-I and DBA as specific marker lectins for the cornified stratified squamous epithelium cells of the Pacific white-sided dolphin. The stomachs of cetaceans and terrestrial mammals have similar histological features and mucous glycoconjugate content.     

Complex Carbohydrates Occurring in the Digestive Apparatus of Umbrina Cirrosa (L.) Fry

The glycosoaminoglycans in the digestive apparatus of immature fish have important biological functions and are involved in morphofunctional differentiation. The aim of this study was to investigate the glycoconjugate histochemistry in the different parts of the digestive apparatus (oesophagus, stomach, intestine) of Umbrina cirrosa (L.) fry using classical histochemical reactions (periodic acid-Schiff, Alcian blue pH 2.5, Alcian blue/periodic acid-Schiff, high iron diamine, low iron diamine) in conjunction with glycolytic digestions that degrade different classes of glycosoaminoglycans. No differences were observed in the reactivity to conventional histochemical staining of the oesophagus, stomach or intestine among 27-, 34- or 44-day-old fry. In the oesophagus, the mucopolysaccharides contained chondroitin sulphates B and A and/or C, heparan sulphate and chondroitin. In the stomach, only neutral glycoconjugates were revealed, whereas in the intestine there were only chondroitin sulphates. Some differences in the type and content of glycoconjugates were found in Umbrina cirrosa (L.) fry compared to those of adult subjects, probably related to different dietary habits and to changes in the environmental conditions.     

Complex carbohydrate histochemistry and ultracytochemistry of the sheep lacrimal gland

The chemical content of the secretion of the sheep lacrimal gland was analysed at the light and electron microscope levels by applying histochemical techniques and an ultrastructural histochemical method (periodic acid, thiocarbohydrazide and silver proteinate). Mucosubstance histochemistry demonstrated acidic glycoconjugates, mainly sulphated, in the mucous and seromucous glandular cells and in the apical portion of the cells lining the terminal ducts. Moreover, secretory granules, stained with PA-TCH-SP, showed a different localization of the reaction product. The presence of lysozyme was also found in the glandular serous cells. These histochemical studies demonstrate that the secretion of sheep lacrimal glands is mixed, having serous, mucous and seromucous components, and that an excellent correlation exists between the secretory granule substructure and glycoprotein localization.     

Glandular structures in the major interlobular and extrapancreatic ducts in the guinea pig

The morphology of the pancreatic duct system did not receive much attention as compared to the microanatomy of the exocrine and endocrine pancreas. The histological peculiarities of the excretory duct system are of major importance especially in laboratory animals like guinea pigs. The paper describes the histological peculiarities of the major interlobular and extrapancreatic ducts in guinea pigs. The pancreatic tissue samples were collected from five guinea pigs. For histological investigation, several pancreatic fragments underwent fixation in 10% buffered formalin and were later processed by the standard paraffin technique. Subsequentially, tissue sections were stained by Goldner's trichrome staining. The mucous substances were assessed by Alcian blue and Periodic acid–Schiff staining methods. The interlobular and main pancreatic ducts in the guinea pig present a simple columnar epithelium surrounded by a thick layer of dense connective tissue. The aforementioned epithelium of the main pancreatic ducts includes principal cells, goblet cells and basal cells. Additionally, the ductal epithelium presents occasional unicellular multiloculated intraepithelial mucous glands and prominent extraepithelial glands. The latter adopts a simple or compound tubular feature. The mucus elaborated by the three glandular types is mostly neutral in goblet cells, predominantly acidic in extraepithelial ductal glands, and a similar amount of acidic and neutral mucin in intraepithelial glands. In conclusion, the epithelium-associated mucous glands in the interlobular and main pancreatic ducts in the guinea pig are restricted not only to goblet cells. A substantial mucous discharge probably with a protecting role against irritative pancreatic juice derives from the main ductal intraepithelial and extraepithelial glands.     

Die Entwicklung der Magendrüsen der Katze 1 (Felis silvestris catus)

The development of the gastric glands of the cat (Felis silvestris catus) There are five stages in the development of the cat's gastric glands: 1. During the stage of the indifferent epithelium from day 19 to day 24, the anlage of the stomach develops with all layers; 2. The stage of gland formation from day 24 to day 41 is the beginning of the gland buds. They develop in connection with endocrine cells on day 34 into primitive oxyntic and primative mucous cells. The latter form the basis for all other cells, including the surface mucous cells; 3. During the stage of gland evagination from day 42 to 55, the anlagen are separated into primitive pits and tubules, while the cells continue to differentiate and the first intermediate cells are seen; 4. The stage of gland branching from day 56 to birth is characterized by the formation of additional glands at the bottom of the pits which change the ordinary anlagen into branched glands. During this stage, the cardiac glands are formed; 5. In the stage of gland maturation from birth to the 9th week, the peptic cells are formed and the glands start functioning. The oxyntic cells show carbonic-anhydrase activity and signs of acid secretion, and, between the weeks 4 and 8, the peptic cells contain pepsinogen, producing a negative reaction to PAS and a positive reaction to HID. Mucous cells and mucous neck cells produce PAS- and AB-positive mucin.     

Ultrastructural study of sheep lacrimal glands.

Sheep lacrimal glands are mixed glands, consisting of tubulo-acinar units succeeded by ducts of simple morphology. The secretory portions consist of three cell types: mucous, seromucous and serous, which may be intermingled in the same acinus or may form acini wholly made of only serous or mucous cells. Mucous cells show a rough endoplasmic reticulum that is reduced to a few cisternae located near the cell base and among the interstices of the secretory droplets. Mucous granules appear uniformly electron-lucent. Serous cells display a typical structure; serous granules can be uniformly electron-dense or composed of dense inclusions dispersed in an electron-lucent matrix. The seromucous granules have a bizonal substructure: a dense core is embedded in a highter matrix. Secretory acini are succeeded by intercalated ducts; the epithelium of these ducts gradually increases in height to form a kind of excretory duct, without the intervention of striated ducts.     

Light and electron microscopic studies of the trachea in the one-humped camel (Camelus dromedarius)

Histology of trachea of camel (Camelus dromedarius) was studied using light, scanning (SEM) and transmission electron microscopy (TEM). Tissue samples taken from the trachea (proximal, middle and distal part) were routinely prepared for histology (LM, EM) and stained with haematoxylin and eosin, Van Giesson (VG), Alcian blue, Periodic acid schiff (PAS), Masson's trichrome (MT), Verhof, PAS-VG and PAS-MT. The trachea of camel consists of 66-75 incomplete cartilaginous rings of hyaline. The lamina epithelium is composed of pseudostratified-ciliated columnar epithelium with many goblet cells. Submucosal layers were loose connective tissue with many elastic fibres. The mucosal and submucosal layers were 517.2 +/- 61.6 microm (n = 20) thick. Submucosal glands were tubuloalveolar with mucous (acidic and neutral) secretions. Trachealis muscle was attached to the inside sheet of tracheal cartilage. Ultrastructural studies showed that surface epithelium is pseudostratified with mucus-producing goblet cells, ciliated and basal cells, similar to other mammals. The ciliated cells contained many mitochondria, oval nucleus and many big granules. In scanning electron microscopy (SEM) studies, viscoelastic layers were observed on the epithelial surface of trachea, and there were highly condensed cilia under this layer.     

Histological and histochemical study of the proventriculus (Ventriculus glandularis) of common starling (Sturnus vulgaris)

The histological and histochemical structures of the proventriculus of starling (Sturnus vulgaris) were examined using haematoxylin and eosin and special staining, that is periodic acid schiff (PAS), Masson's trichrome, Alcian blue, Orcein and Reticulin. All three cranial, middle and caudal parts of the proventriculus were also studied. The study results showed that the wall of the proventriculus consisted of mucosal, submucosal, muscular and serosal tunics. The mucosal tunic presented folds and sulci on its luminal surface. In the first third of the proventriculus, the tunica mucosa characterized by presence of folds lined by stratified squamous epithelium and presence of simple tubular glands in the lamina propria. In the middle and caudal thirds of the proventriculus, the surface was covered by a columnar epithelium, and the branched tubular glands were extended through the lamina propria. From the base of the branched tubular glands, the deep proventricular glands were observed that were compound tubuloalveolar lobules. The surface epithelium of the tunica mucosa and the cells lining the proventricular glands showed a positive reaction to PAS and Alcian blue stainings. In addition, the epithelial cells of the tubular and branched tubular glands showed Masson's trichrome-positive reaction. The submucosal tunic was thin in the proventricular wall. The tunica muscularis was formed by a thin inner layer of longitudinal smooth muscle fibres and a thick outer layer of circular fibres. The serosa consisted of loose connective tissue, rich in blood vessels and covered by mesothelium.     

Normal ultrastructure and histochemical characteristics of canine lacrimal glands

Lacrimal glands of 12 dogs free of ocular disease were examined to determine the normal structure of these glands. The glands consisted of tubuloacinar cells that ultrastructurally and histochemically were of a single type of secretory cell in the tubules and possibly 3 types of secretory cells in the acini. The tubular epithelium contained homogenous electron-dense granules that stained as neutral glycoconjugates (periodic acid-Schiff positive and Alcian blue and high iron diamine negative). The predominant acinar cells contained granules of lesser electron density than those of the tubules, and stained as sialomucin (Alcian blue [pH 2.5] and periodic acid-Schiff-positive, and high iron diamine-negative). A second type of acinar cell was in peripheral lobules that ultrastructurally and histochemically appeared like lipid granules (positive with oil red O and osmium tetroxide). Ultrastructurally, a third type of acinar granule was finely granular, electron-lucent, and frequently coalesced. It was not readily apparent whether the latter was an artifact, a stage in the maturation of the sialomucin granules, or a third type of acinar granule. Individual acinar cells usually had a predominance of 1 granule type, but greater than 1 granule type could be found in some cells. The basal surfaces of the acinar, tubular, and ductal cells were incompletely ensheathed by myoepithelial cells. Plasma cells, lymphocytes, mast cells, endothelial cells, fat cells, and Schwann cells composed the cellular elements of the interstitium. Lymphocytes, mast cells, and nerve endings also were found in the parenchyma.     

Gill epithelium of an angler catfish,Chaca chaca (Siluriformes,Chacidae): Enzyme and glycoprotein histochemistry

A series of histochemical techniques have been employed to localize alkaline phosphatase, acid phosphatase, non-specific esterase, catalase and peroxidase; and to visualize and characterize glycoprotein (GPs) moieties in the epithelium of gill arch, gill filaments and secondary lamellae of an angler catfish Chaca chaca. The epithelium of gill arch and gill filament shows strong alkaline phosphatase activity in the deeper layer epithelial cells; strong non-specific esterase activities in the outer layer epithelial cells; and weak acid phosphatase activity throughout the epithelium. The activity of these enzymes in the secondary lamellae is weak. The catalase and peroxidase show strong activities in the blood cells of the secondary lamellae. Various classes of GPs have been identified and characterized in the mucous secretions of the gill epithelium of C. chaca. These include—GPs with oxidizable vicinal diols, GPs with sialic acid residues without O-acyl substitution and GPs with O-sulphate esters. The functional significance of different enzymes in gill epithelium and the GPs in the mucus secreted on the surface has been discussed with the physiology of the gills in relation to the characteristic habit and habitat of the fish.     

铅暴露对黄河鲤鱼黏液细胞数量和类型的影响

利用阿新蓝过碘酸雪夫染色方法,研究了铅浸泡对鲤鱼黏液细胞的影响。结果表明,铅暴露对于黏液细胞数量的影响是先增后降,方差分析表明,Pb2+0.01、0.1 mg/L组口腔上皮和后肠,0.1 mg/L组鳃丝,以及所有试验组前肠和中肠黏液细胞密度与对照组相比均显著增加(P<0.01或P<0.05),而2 mg/L和5 mg/L组口腔上皮和后肠,0.01、2、5 mg/L组鳃丝上黏液细胞密度与对照组相比均无显著性差异(P>0.05)。铅对于黏液细胞类型的影响是可以促进Ⅱ型黏液细胞,抑制Ⅰ、Ⅲ、Ⅳ型黏液细胞的形成。     

Necrotic processes in the gill tissue of carp caused by changes in the aquatic environment

Histological findings show that various pathogenic processes are involved in the cases of clinically detected branchionecrosis. The changes in gills occurring at toxic branchionecrosis are a necrobiotic process in their nature; in gill lamellae this process manifests itself as a swelling of the respiratory epithelium and as dilation of lamellar sinuses, and in stratified epithelium of gill arch and filaments it takes on the form of the hypertrophy and hyperplasia of epithelial cells. In severe cases the swollen epithelium forms deposits filling up also the interlamellar space. Toxic branchionecrosis is a term referring to a disease when the pathological processes are caused by ammonia intoxication and autointoxication, the main role being played by pH value. At experimental ammonia intoxication the changes in gills are about the same as in toxic branchionecrosis but the symptoms are not so pronounced. Changes characteristic of spherosporosis involve severe necrotic processes in the respiratory epithelium as well as in the stratified epithelium of gills, and the epithelium harbours different developmental stages of spherospores. The spherospores are easily detected when stained with alcian blue at pH 2.5. The changes in the gills of carp naturally invaded by spherospores with additional ammonia intoxication correspond to findings characteristic of toxic branchionecrosis: degeneration of stratified and respiratory epithelium of gills combined with necrotic changes caused by the developmental stages of spherospores. When water is highly oxygenated with chlorinated lime, large cells with eosinophilic granules severely multiply mainly in the stratified epithelium of gills; this process is considered as an inflammatory process. Parasitic invasions by Trichodina and Chilodonella induce local inflammatory reactions with the swelling of the respiratory epithelium.     

The Fine Structure of the Fundic Stomach Epithelium of the Western Newt

The effects of varying intensities of histamine stimulation on cells of the western newt fundic stomach epithelium have been studied. Oxyntic, mucous neck and surface mucous cells all secrete. In the oxyntic cell, the main effects are a movement of zymogen granules toward the apex, the discharge of their contents into the lumen and, eventually, their disappearance from the cytoplasm. Additionally, the tubular membrane elements (tubulo- vesicles) fuse with the apical plasma membrane with a corresponding reduction in their numbers and an increase in microvilli, which eventually are so numerous as to virtually occlude the lumina of gastric glands. Total histamine stimulation results in apparent damage to the oxyntic cells. Mucous neck and surface mucous cells release their mucigen droplets by exocytosis and are grossly changed in appearance also.     

Lectin Histochemistry of Glycoconjugates in Horse Salivary Glands

The glycoconjugate content of major horse salivary glands was investigated by means of horseradish peroxidase-conjugated lectins. Qualitative differences were observed in the terminal sugar residues of secretory glycoproteins and glycoconjugates linked to the apical surface of excretory duct epithelial cells. Mucous acinar cells in mandibular and sublingual glands contained oligosaccharides with D-galactose, α- and β-N-acetylgalactosamine, N-acetylglucosamine and fucose residues, whereas mandibular, sublingual and parotid serous cells contained only oligosaccharides with terminal α- and β-N-acetylgalactosamine residues. The apical portion of striated and interlobular duct lining cells of mandibular and sublingual glands stained for α- and β-N-acetylgalactosamine and for N-acetylglucosamine. In parotid gland the cytoplasm of intercalated duct cells and the apical surface of striated duct epithelial cells stained for α-N-acetylgalactosamine.