Mucous Cells in Micropogonias furnieri gills: Histochemistry and Ultrastructure

The characteristics of the mucous cells located in the gills of the fish Micropogonias furnieri were investigated. Using histochemical procedures that included methods for localization and characterization of glycoproteins (GPs), no differences were detected between the mucous cell contents of the primary and secondary lamellae. The GPs were identified with (a) oxidizable vecinal diols; (b) sialic acids and some of their chain variants, C7 or C9; (c) carboxyl groups and (d) sulphate groups. The electron microscope showed large mucous globules of different electro densities from mucous cells located deep in the epithelium between the other epithelial cells; the release of mucus by exocytosis was observed. GPs secreted on the surface of the mucous cells was suggested to be important for the lubrication, protection and inhibition of microorganisms. It is possibility that GPs could have similar roles in Micropogonias furnieri gills.     

Lateral musculature in the whitemouth croaker (Micropogonias furnieri): its characterization with respect to different gonadal conditions

Histochemical and ultrastructural studies were performed on the lateral musculature from individual female whitemouth croaker, Micropogonias furnieri, at the anterior, medium and posterior regions. Based upon histochemical myosin-ATPase (m-ATPase) determination, diverse types of red, pink and white fibres were discerned. Red muscle had abundant mitochondria and stained intensely for aerobic enzymes, white muscle scarcely stained for the same enzymes and pink muscle responded in an intermediate manner. Both white and pink muscle had few mitochondria. The relative proportion of red muscle increased towards the caudal region; pink muscle diminished towards this region and white muscle modified its proportion only in the anterior region. m-ATPase activity showed differences in relation to the gonadal condition along the body, particularly in the white fibres at the anterior and medium regions.     

The mucosa of the digestive tract in Micropogonias furnieri: a light and electron microscope approach

The histomorphological aspects as well as the histochemical content and distribution of glycoproteins (GPs) in the mucosa of the digestive tract of the white croaker Micropogonias furnieri were studied. The buccopharyngeal cavity and the esophagous showed a squamous stratified epithelium with mucous cells. The stomach presented three portions: cardias, fundus and pylorus. Tubular glands formed by a single type of gland cell were located along the cardias and fundus. Histochemical tests showed that the buccopharyngeal cavity and the esophagous presented the largest amount of the different types of mucosubstances. Both organs showed abundant secretory mucous cells that synthesize large quantities of neutral, sulphated and sialylated GPs. The surface epithelium in the cardias and fundus synthesized and secreted scarce sialylated and neutral GPs whereas the secretions of the apical surface were abundant. The pylorus secreted large amounts of neutral as well as sulphated and sialylated GPs. Gland cells secreted neutral GPs. The ultrastructural features of the gut cells were quite similar to those of other teleosts. The buccopharyngeal cavity and the esophagous surface epithelial cells, identified by their superficial localization, were characterized by cytoplasmic vesicles of different size. Abundant goblet cells with secretory mucous granules were also present. Gastric glands in the stomach contained just one form of cell with a fine structure similar to cells that secrete pepsinogen.     

A histochemical study of the camel (Camelus bactrianus) duodenal glands

The complex carbohydrates in the camel duodenal glands were examined histochemically at light and electron microscopic levels. The duodenal glands of the camel were distributed in the submucosa 2 m caudal from the pylorus. These were branched tubuloalveolar glands. The terminal portion of each lobule was formed by only one type of mucous cell. The duodenal gland cells contained acidic and neutral carbohydrates. The mucous cells mainly contained sulfate and carboxyl carbohydrate with sialic acid, and they also contained a few neutral carbohydrates with different saccharide residues such as mannose, glucose, galactose, N-acetyl glucosamine and N-acetyl galactosamine. The results showed that the secretary granules of the duodenal glands in the camel contain mainly acidic carbohydrates. These findings seem to be the morphological characteristics of the duodenal glands in the camel.     

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.     

Ultrastructure features of the surface of the gills and the lower pharyngeal jaw of the Tilapia Zilli,redbelly tilapia (Coptodon Zillii,Gervais, 1848)

The present investigation was designed to describe the surface ultrastructure of the gill system of tilapia Zilli. The gill system is formed from four gill arches and each gill arch carries a row of gill filaments on its convex border and two rows of the gill rakers on its concave border. The quadrilateral interbranchial septum has elevated part at the level of the third gill arch. By SEM observations, the gill arch was divided into three regions: rostral, middle and caudal region. The caudal region contained two characteristic structures: oval leaf‐like structure and rounded‐shaped structure. Each oval leaf‐like structure carried two lateral rows of the triangular pointed spines separated by a median groove. All surfaces of gill arches, rakers and filaments were covered with a mosaic of the polygonal pavement cells, in addition to the opening of chloride cells and mucous cells. The gill arch and gill raker had only one appearance of taste buds named type I. Meanwhile, the filaments contained two types of different appearance of the taste buds named: type I and type II. Type I was the main common and similar to that present in gill arch and raker and characterized by its blunt end, while type II had hair‐like structures that projected from the volcano‐shaped depression. The gill rakers were formed from central axis surrounded by two lateral lobulated regions which carry pointed spines, taste buds and the opening of chloride cells. The surface of triangular lower pharyngeal jaw carries numerous teeth‐like papillae which originated from the socket‐like depression.     

A lectin histochemical study on the testis of the babirusa, Babyroussa babyrussa (Suidae)

The distribution of lectin bindings in the testis of babirusa, Babyrousa babyrussa (Suidae) was studied histochemically using 10 biotinylated lectins, Peanut agglutinin (PNA), Ricinus communis agglutinin (RCA I), Dolichos biflorus agglutinin (DBA), Vicia villosa agglutinin (VVA), Soybean agglutinin (SBA), Wheat germ agglutinin (WGA), Lens culinaris agglutinin (LCA), Pisum sativum agglutinin (PSA), Concanavalin A(Con A) and Ulex europaeus agglutinin (UEA I). Nine of 10 lectins showed a variety of staining patterns in the seminiferous epithelium and interstitial cells. The acrosome of Golgi-, cap- and acrosome-phase spermatids displayed various PNA, RCA I, VVA, SBA and WGA bindings, indicating the presence of glycoconjugates with D-galactose, N-acetyl-D-galactosamine and N-acetyl-D-glucosamine sugar residues respectively. No affinity was detected in the acrosome of late spermatids. LCA, PSA and Con A which have affinity for D-mannose and D-glucose sugar residues were positive in the cytoplasm of spermatids and spermatocytes. DBA was positive only in spermatogonia. In addition to DBA, positive binding in spermatogonia was found for VVA, WGA and Con A, suggesting the distribution of glycoconjugates with N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, D-mannose and D-glucose sugar residues. Sertoli cells were stained intensely with RCA I, WGA and Con A. In Leydig cells, RCA I and Con A were strongly positive, while WGA, LCA and PSA reactions were weak to moderate. The present findings showed that the distribution pattern of lectin binding in the testis of babirusa is somewhat different from that of pig or other mammals reported previously.     

Histological, histochemical and immunohistochemical study of the lymph nodes of the one humped camel (Camelus dromedarius)

Prescapular, femoral, mesenteric, mediastinal and splenic lymph nodes from nine camels of one to 12 years of age were studied. There were no obvious structural differences between these different lymph nodes or between the ages. The lymph nodes were surrounded by a capsule formed of two layers, an outer thicker layer of connective tissue and an inner thinner layer mainly of smooth muscles. Trabeculae extended from the inner layer of the capsule dividing the parenchyma characteristically into incomplete lobules. Subcapsular and trabecular lymphatic sinuses were supported by a reticular fiber network. The parenchyma was uniquely different from that of other species, as it was arranged in the form of lymphoid follicles and interfollicular lymphoid tissue. The lymphoid follicles of CD22 positive lymphocytes were supported by a reticular fiber network. This fine network of α-smooth muscle actin positive cells enclosed the lymphoid follicles. The interfollicular tissue was mainly made up of diffusely distributed CD3 positive lymphocytes. MHC class II: DR was expressed by most lymphocytes of the follicles and interfollicular tissue. Lymphatic sinuses and high endothelial venules were found in the interfollicular zone. The lymphatic sinuses were lined by discontinuous endothelial cells. The wall of the high endothelial venules was infiltrated by several lymphocytes and enclosed in a layer of α-smooth muscle actin positive cells. Acid phosphatase positive cells were evenly distributed in the interfollicular zone. A few cells were localized in the lymph follicles. Alkaline phosphatase was observed in the endothelium of the lymphatic sinuses and in the lymphoid follicles.     

A mucinous histochemical study on malignancy of aberrant crypt foci (ACF) in rat colon

The relationship between malignancy and number of crypts (crypt multiplicity) comprising aberrant crypt foci (ACF) was investigated, by studying changes in the mucous nature of ACF with 5 crypts or less, ACF with 6-13 crypts, adenomas and invasive adenocarcinomas induced by 1,2-dimethylhydrazine in distal colon of rats. A paradoxical Con A-staining was performed for goblet cell mucins. Of the sulfomucin-dominant ACF with 1-3 crypts, 82.6% had labile class III mucin, similar to the distal colon in the normal rats. However, in most of the goblet cell mucin produced by the ACF with 4-5 crypts with an indicated relation to colorectal carcinoma or the sialomucin (SiM) -dominant ACF with 1-3 crypts, mucin types other than class I were rarely present. The incidence of class I mucin decreased with the increase in crypt multiplicity of ACF or in the degree of histological malignancy, with the lowest incidence of 40% in adenocarcinomas. In contrast, the incidence of class II mucin increased markedly with the increase in crypt multiplicity of ACF or in the degree of histological malignancy, with the highest incidence in adenocarcinomas (95%). The ACF with 6-13 crypts had a mucous profile similar to that of adenomas. These results suggested that malignancy of ACF related to the crypt multiplicity. In the ACF with 1-3 crypts, SiM-dominant ACF had the potential to progress to malignant lesions.     

Histochemical detection of glycoconjugates in the inner perivitelline layer of Japanese quail (Coturnix japonica)

The avian inner perivitelline layer (IPVL), a homologous structure to the mammalian zona pellucida, is deposited between the granulosa cells and the oocyte cell membrane during folliculogenesis. In this glycohistochemical study, a panel of fluorescein isothiocyanate (FITC)-labelled lectins was used to characterise and localise the oligosaccharide sequences of the IPVL glycoproteins at different stages of follicular development in the quail ovary. Deacetylation and sialidase digestion were also performed prior to lectin cytochemistry. Contrary to mammals, where the topographical distribution of these carbohydrates is not uniformly distributed throughout the zona pellucida, indicating the regionalisation of oligosaccharide chains, our results demonstrated a homogenous lectin staining of the comparatively thin IPVL. We also found variations in the presence and distribution of the carbohydrate residues in the IPVL during different stages of follicular growth. The IPVL of pre-vitelline follicles distinctly stains with WGA, sWGA and SBA, demonstrating the presence of D-GlcNAc, Neu5Ac and α-D-GalNac in the glycoproteins of the forming IPVL. No staining was found with ConA (specific for α-D-Man, α-D-Glc), LCA (α-D-Man, α-D-Glc), PNA (β-D-Gal-(1-3)-D-GalNAc, VAA (Gal), DBA (α-D-GalNAc(1-3)-GalNAc and UEA-I (α-L-Fuc). With continuing follicular growth of the oocyte and the follicle, this staining pattern changed. LCA and PNA-staining in the IPVL became distinctly positive. As the IPVL of immature oocytes distinctly stains with WGA/sWGA-FITC, but spermatozoa do not bind to immature zona, it appears questionable that carbohydrate residues detected by WGA/sWGA play a major role in sperm-IPVL binding, as suggested in previous investigations.     

日本七鳃鳗血细胞显微及亚显微结构

用光镜和透射电镜技术对１０尾日本七鳃鳗的血细胞进行了观察。结果表明：红细胞为圆形或椭圆形,胞质不很均匀,电子密度较低,含有板状嵴线粒体,核呈圆形或椭圆形。中性粒细胞表面有伪足,形态各异,胞质内含有Ａ、Ｂ２种颗粒（Ａ型颗粒为圆形,电子密度较低;Ｂ型颗粒为长椭圆形,电子密度较高）。嗜酸粒细胞数量较少,胞质内含有Ａ、Ｂ、Ｃ３种形态各异的有膜颗粒（Ａ型颗粒为圆形,电子密度较高,均质;Ｂ型为长椭圆形,颗粒中央有电子密度较高的长椭圆形致密芯;Ｃ型内含有电子密度较高的杆状致密芯,位于颗粒一侧）。单核细胞表面有伪足,胞质内含有粗面内质网、板状嵴线粒体、溶酶体和液泡。淋巴细胞为圆形,核较大,胞质内含有板状嵴线粒体、粗面内质网和嗜天青颗粒。血栓细胞体积较小,形态各异,胞质内含有线粒体、粗面内质网和细小颗粒。     

Ultrastructure of Sarcocystis tenella (Sarcocystis ovicanis)

The ultrastructure of the parasitophorous cyst, the metrocytes and merozoites of Sarcocystis tenella (S. ovicanis) was studied. The general ultrastructural features of the cyst wall and the cells of the protozoan, previously described by other authors, were confirmed. The vesicle-like invaginations of the unit membrane (primary wall) of the cyst are identical with the micropinocytic pits of other animal cells, and we believe that they contribute to the nutritional process of the cyst. The Golgi apparatus must be related to the granular endoplasmic reticulum, since the first cisterna of the curved side of the stack appeared to originate from it. The structure of the micropore was similar to that previously described by other workers, the only difference being that in this study, material was frequently observed inside the lumen. This observation led us to support the previously suggested nutritive role. In addition, crystalloid material limited by cytoplasmic membrane was often observed, but the manner of its formation and role remain unknown.     

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.     

Ultrastructure of Camel (Camelus dromedarius) Spermatozoa

Spermatozoa collected from the caudae epididymidis of six camels were examined by phase contrast, scanning electron and transmission electron microscopy. A comparison of the morphology of the camel spermatozoa with that of other domestic animals is presented.     

马鹿精子形态与超微结构研究

文中对马鹿(Cervus elaphus)精子形态与超微结构进行了研究。马鹿精子经固定、脱水、置换、包埋和聚合,用超薄切片机切片,再用醋酸双氧铀、柠檬酸铅染片,最后于透射电镜下观察其超微结构的变化。观察结果表明:马鹿精子全长(58.75±2.35)μm,由头部(8.93±0.24)μm、颈部(1.00±0.16)μm和尾部(48.18±1.18)μm三部分组成;头部呈扁卵圆状,绝大部分被浓缩且电子密度较高的精核所占据,顶体似帽状扣在精核之上,约占头部的2/3,其前部较为膨大;颈部位于头部和尾部之间,这个区域较短;尾部可分为中段、主段和终段。马鹿尾部轴丝的结构类型为"9+2";微管结构类型为"9+9+2"。     

The effect of ragwort (Senecio jacobea) on the liver of the domestic fowl (Gallus domesticus) : a histopathological and enzyme histochemical study.

1. Twenty-five 1-week-old male chicks were fed for 6 weeks on a standard diet incorporating 7% dried and ground ragwort (Senecio jacobea). The chicks received the standard diet for a further 6 weeks whilst a control group of 25 similar birds received the standard diet throughout. 2. Two birds from each group were killed at intervals of about 1 week and their livers were examined histologically and histochemically. 3. Ragwort feeding caused megalocytosis, focal necrosis, focal hyperplasia and portal fibrosis. There were no veno-occlusive changes. 4. As the lesion progressed there was an overall loss of enzyme activity especially in the areas of necrosis. The groups of small hyperplastic cells showed normal or increased activity and there was increased enzyme activity in the megalocytes. 5. The development of groups of proliferating cells may be a prelude to the neoplastic changes described in earlier reports of longer term studies of the effects of pyrolizidine alkaloids on the liver of chicks.     

Liver Enzymes and Ultrastructure in Rabbit Haemorrhagic Disease (RHD)

Rabbit haemorrhagic disease (RHD) is caused by a calicivirus infection that kills most adult rabbits 24–72 h after viral inoculation. Two liver enzymes (AST, aspartate aminotransferase, and ALT, alanine aminotransferase) were monitored in blood samples of calicivirus-infected rabbits during the short course of RHD. Values of AST were used to differentiate three stages of hepatocellular degeneration in RHD: mild (up to 20-fold increase in AST), moderate (150–200-fold elevation of AST) and severe (more than 1000-fold elevation in AST). Liver samples of rabbits from these three biochemical stages of hepatocellular degeneration of RHD were studied by transmission electron microscopy to define the fine structure of the hepatocytes. In the mild hepatocellular degeneration there was proliferation (microvesiculation) of the smooth endoplasmic reticulum and swelling of mitochondria into spheroid bodies with loss of cristae. In moderate hepatocellular degeneration, vacuolization of cytoplasm and mitochondrial damage continued to be present, and there was also formation of autophagic vesicles. In the severe hepatocellular degeneration of RHD, the altered mitochondria also showed loss of density of their matrix; rupture of cytoplasmic vacuoles led to the formation of large vesicles. Marked depletion of liver glycogen was also found in this late stage of RHD. These data offer a correlation between biochemical and cytological features of the liver during the hepatocellular degeneration of RHD.     

Characterization of glycoconjugates and sialic acid modification in the olfactory bulb of the Chinese fire-bellied newt (Cynops orientalis)

Diverse glycoconjugates are expressed in the vertebrate olfactory bulb and serve as guidance cues for axons of nasal receptor neurons. Although the involvement of glycoconjugates in the segregation of the olfactory pathway has been suggested, it is poorly understood in salamanders. In this study, lectin histochemistry was used to determine glycoconjugate distribution in the olfactory bulb of the Chinese fire-bellied newt (Cynops orientalis). Succinylated wheat germ agglutinin (sWGA), Ricinus communis agglutinin-I and Lens culinaris agglutinin showed different bindings in the nerve fibre layer or glomerular layer, or both, between the main and accessory olfactory bulbs. We then investigated the lectin-binding pattern after the removal of terminal sialic acids using neuraminidase. Desialylation resulted in a change in the binding reactivities with seven lectins. Wheat germ agglutinin, sWGA, soybean agglutinin (SBA) and peanut agglutinin showed different degrees of binding between the main and accessory olfactory bulbs. In addition, SBA showed a heterogeneous labelling of glomeruli in the rostral region of the main olfactory bulb. Our results suggest that terminal sialic acids mask the heterogeneity of glycoconjugates in the olfactory bulb of C. orientalis.