Light and electron microscopy studies of the oesophagus and crop epithelium in Aplysia depilans (Mollusca, Opisthobranchia)

The oesophagus and crop epithelium of Aplysia depilans consist in a single layer of columnar cells with apical microvilli, and some of them also possess cilia. Cell membrane invaginations, small vesicles, multivesicular bodies and many dense lysosomes were observed in the apical region of the cytoplasm. In most cells, a very large lipid droplet was observed above the nucleus and a smaller one was frequently found below the nucleus; glycogen granules are also present. Considering these ultrastructural features, it seems that these cells collect nutritive substances from the lumen by endocytosis, digest them in the apical lysosomes and store the resulting products. The cell bodies of mucus secreting flask-shaped cells are subepithelial in the oesophagus and intraepithelial in the crop. Histochemistry methods showed that the secretion stored in these cells contains acidic polysaccharides. Secretory vesicles with thin electron-dense filaments scattered in an electron-lucent background fill most of these cells, and the basal nucleus is surrounded by dilated rough endoplasmic reticulum cisternae containing small tubular structures. Considering the relatively low number of secretory cells, mucus production cannot be high. Moreover, since protein secreting cells were not observed in either oesophagus or crop, extracellular digestion in the lumen of these anterior segments of the digestive tract most probably depend on the enzymes secreted by the salivary and digestive glands.     

Light and electron microscopic study of the anterior oesophagus of Bulla striata (Mollusca,Opisthobranchia)

Lobo‐da‐Cunha, A., Oliveira, E., Alves, Â., Coelho, R. and Calado, G. 2010. Light and electron microscopic study of the anterior oesophagus of Bulla striata (Mollusca, Opisthobranchia). —Acta Zoologica (Stockholm) 91 : 125–138. The anterior oesophagus of Bulla striata was investigated with light and electron microscopy. In the most anterior region, the ridges of the oesophageal wall are covered by a ciliated columnar epithelium forming large apical blebs which are released into the lumen, an activity that is particularly intense in the oesophageal pouch. In the last two‐thirds of the anterior oesophagus, the epithelium is covered with microvilli embedded in a cuticle, but apocrine secretion and cilia are absent. Subepithelial secretory cells are very abundant in the oesophageal wall, except in the roof of the pouch. They have a long neck that crosses the epithelium, whereas the cell body containing the nucleus is embedded in the connective tissue. Large electron‐lucent secretory vesicles and many Golgi stacks fill most of their cytoplasm. The histochemical and cytochemical assays show that these cells secrete acid mucopolysaccharides. With the current and future studies we aim to obtain data for the establishment of relationships between morphofunctional features of the digestive system and food types in cephalaspideans. Additionally, the new data about the oesophageal pouch of B. striata may be useful for the establishment of eventual homologies with the oesophageal diverticula of other opisthobranchs.     

Histology and ultrastructure of the salivary glands in Bulla striata (Mollusca, Opisthobranchia)

Abstract. The ribbon‐shaped salivary glands in Bulla striata were studied with light microscopy and transmission electron microscopy (TEM). Secretion is produced in tubules formed by two types of secretory cells, namely granular mucocytes and vacuolated cells, intercalated with ciliated cells. A central longitudinal duct lined by the same cell types collects the secretion and conducts it to the buccal cavity. In granular mucocytes, the nucleus is usually central and the secretory vesicles contain oval‐shaped granular masses attached to the vesicle membrane. Glycogen granules can be very abundant, filling the space around the secretory vesicles. These cells are strongly stained by PAS reaction for polysaccharides. Their secretory vesicles are also stained by Alcian blue, revealing acidic mucopolysaccharides, and the tetrazonium reaction detects proteins in minute spots at the edge of the vesicles, corresponding to the granular masses observed in TEM. Colloidal iron staining for acidic mucopolysaccharides in TEM reveals iron particles in the electron‐lucent region of the vesicles, while the granular masses are free of particles. In vacuolated cells, which are thinner and less abundant than the granular mucocytes, the nucleus is basal and the cytoplasm contains large electron‐lucent vesicles. These vesicles are very weakly colored by light microscopy techniques, but colloidal iron particles could be observed within them. The golf tee‐shaped ciliated cells contain some electron‐dense lysosomes in the apical region. In these cells, the elongated nucleus is subapically located, and bundles of microfibrils are common in the slender cytoplasmic stalk that reaches the basal lamina. The morphological, histochemical, and cytochemical data showed some similarities between salivary glands in B. striata and Aplysia depilans. These similarities could reflect the phylogenetic relationship between cephalaspidean and anaspidean opisthobranchs or result from a convergent adaptation to an identical herbivorous diet.     

Light and electron microscopy of the leucocytes of Crassostrea virginica (Mollusca: Pelecypoda)

Summary The fine structure of oyster leucocytes resembles to a great extent, that of typical eucaryotic cells. Organelles which have been described for the first time in this report are light granules, dense granules, protocentriole and X structure. Light microscopy reveals two morphological types of oyster leucocytes: agranular and granular. Based upon nuclear morphology and cytoplasmic compositions revealed in electron microscopy, at least three types of agranular and one type of granular cells are recognized.In the Giemsa-stained preparations, granular leucocytes exhibit three distinct types of cytoplasmic granules: refractile, dark blue, and pink, which presumably correspond to light granules Type A, B, and C seen in the electron micrographs. A granular leucocyte may contain one or more types of granules. Cytochemical investigations show that oyster leucocytes contain at least three hydrolytic enzymes: non-specific esterases, acid, and alkaline phosphatase. The latter two enzymes constitute 63% of the enzyme activity detected. These intracellular enzymes may be associated with the light granules and/or lysosome-like bodies.It is also demonstrated that the granular leucocyte population is significantly higher (P<0.001) in the oysters experimentally infected with Bacillus mycoides (72.19±4.71%) as contrasted with that of the controls (37.18±4.48%).Leucocytes in progressive stages of degeneration are also described.Contribution No. 71 from Marine Research Laboratory, University of Connecticut.The initial phase of this investigation was carried out at the Department of Zoology, Rutgers, The State University, New Brunswick, New Jersey, and supported by Public Health Service Research Grant AI-00781 from the National Institute of Allergy and Infectious Diseases of the National Institute of Health, awarded to Dr. L. A. Stauber. Supported by a grant from the University of Connecticut Research Foundation and Faculty Summer Fellowship to S. Y. Feng.     

Light and electron microscopy study of the salivary gland secretory cells of Helicoidea (Gastropoda, Stylommatophora)

We have studied the secretory goblet-cells of the salivary gland of six species of Helicoidea: Elona quimperiana, Trissexodon constrictus, Hygromia limbata, Cernuella aginnica, Cepaea nemoralis and Helix aspersa, using light microscopy and transmission electron microscopy. In each of the species studied, we have been able to demonstrate the presence of four cell types. Following a comparative study with the goblet-gland cells present in the six species in question, and the comparison of our data with previously published reports on the entire set of Stylommatophora, we have established the homologies corresponding to the cell types observed. Our hypotheses were based primarily on the morphology of the rough endoplasmic reticulum and of the secretory vesicles. Accordingly, we have defined five cell types for these six species. The first three cell types, A, B and C, appear in the six species studied and seem to be present in all the stylommatophores. In order to standardize the terminology used by the different authors, we propose that these cell types be called: 'swollen RER cisterns mucocyte', 'granular mucocyte' and 'alveolar cell', respectively. The D-cell type or 'basophilous cell' is present only in Hygromia limbata, Cernuella aginnica, Cepaea nemoralis and Helix aspersa. The E-cell type or 'vacuolated cell' appears only in Elona quimperiana and Trissexodon constrictus.     

Ultrastructural and histochemical study of the salivary glands of Aplysia depilans (Mollusca, Opisthobranchia)

The digestive system of the sea hare, Aplysia depilans , includes a pair of ribbon-shaped salivary glands. A central duct and a large blood vessel run close to each other along the length of these glands and both are surrounded by a layer of muscle cells. Three cell types form the glandular epithelium: granular cells, vacuolated cells and mucocytes. The granular cells possess cilia and spherical secretion granules, located primarily in the apical region. The granules of immature cells have a low electron density and are mainly formed by neutral polysaccharides with small amounts of proteins. The granules of mature cells are larger, have a high electron density and are mainly formed by proteins with lower amounts of neutral polysaccharides. Transition stages between immature and mature granular cells are observed. The vacuolated cells are large and frequently pyramidal in shape, but after the application of histochemical techniques almost all vacuoles remain uncoloured. The numerous vacuoles contain flocculent material in a clear background and the mitochondria possess large crystalline structures in the matrix. A pyramidal shape is also typical of the mucocytes, which are filled with vesicles containing granular masses surrounded by a network of secretion material. These large cells are strongly stained by Alcian blue, revealing the presence of acidic mucopolysaccharides. This is the first ultrastructural study of the salivary glands in opisthobranch gastropods.     

Histochemical analysis of glycoproteins in the secretory cells in the gill epithelium of a catfish, Rita rita (Siluriformes, Bagridae)

Glycoproteins (GPs) were visualised histochemically in the secretory cells – the mucous goblet cells (the type A and the type B), the serous goblet cells, the club cells and the epithelial cells in the gill epithelium of Rita rita. The type A mucous goblet cells, the type B mucous goblet cells and the epithelial cells elaborate GPs with oxidizable vicinal diols and GPs with sialic acid residue without O-acyl substitution. In addition, GPs with O-sulphate esters are elaborated by the type A and GPs with O-acyl sugars by the type B mucous goblet cells. GPs are absent in the serous goblet cells and are with oxidizable vicinal diols in low moieties in the club cells. The analysis of the results elucidates interesting differences in the composition and concentration of GPs in the mucus elaborated by the epithelium of the gill arches and the gill rakers; and the gill filaments and the secondary lamellae indicating the potential importance of the glycoproteins at these locations. GPs elaborated on the surfaces of the gill arches and the gill rakers could be associated to assist in feeding activities and on the surfaces of the gill filaments and the secondary lamellae in the respiratory activity.     

The morphology and ultrastructure of salivary glands of Zoraptera (Insecta)

The salivary glands of two species of Zoraptera, Zorotypus caudelli and Zorotypus hubbardi, were examined and documented mainly using transmission electron microscopy (TEM). The results obtained for males and females of the two species are compared and functional aspects related to ultrastructural features are discussed. The salivary glands are divided into two regions: the secretory cell region and the long efferent duct, the latter with its distal end opening in the salivarium below the hypopharyngeal base. The secretory region consists of a complex of secretory cells provided with microvillated cavities connected by short ectodermal ducts to large ones, which are connected with the long efferent duct. The secretory cell cytoplasm contains a large system of rough endoplasmic reticulum and Golgi apparatus producing numerous dense secretions. The cells of the efferent duct, characterized by reduced cytoplasm and the presence of long membrane infoldings associated with mitochondria, are possibly involved in fluid uptaking from the duct lumen.     

Morphological studies on the periostracum of the fresh-water mussel Amblema (uniondae): Light microscopy, transmission electron microscopy, and scanning electron microscopy

The structure of the periostracum in the fresh-water mussel Amblema has been described using light microscopy, transmission elec;ron microscopy, and scanning electron microscopy. The structure and evolutive course of the periostracum was studied along its entire length, from the periostracal groove until it forms the tough outer covering of the shell. At least five structurally and functionally distinct regions were identified. In addition, the periostracum itself was seen to be a multilayered structure consisting of three major layers which are themselves subdivided into minor layers. From these morphological observations, a regulatory role for the various periostracal layers in mineral trapping, nucleation, and the subsequent formation of the prismatic and nacreous layers of the shell can be postulated.     

Ultrastructure of multicellular foregut glands in selected Solenogastres (Mollusca)

Foregut glands in Solenogastres are not only of importance in biological processes like feeding and digestion, but multicellular foregut glands also provide valuable characters for taxonomy and systematics. Here, the fine structure of four different types of foregut glands is investigated: the Meioherpia-type ventrolateral foregut gland of Meioherpia atlantica, the Simrothiella-type ventrolateral foregut gland of Simrothiella cf. margaritacea, and the Pararrhopalia-type ventrolateral foregut gland as well as the dorsal gland of Pararrhopalia pruvoti. Thereby, special focus is set on the arrangement of glandular and supporting cells within the glands, and on the characterization of glandular cells according to the size, shape and electron-density of their secretional vesicles. It is shown that the investigated glands are complex organs composed of non-glandular supporting cells and two to five glandular cell types producing discrete secretions.     

The ultrastructure of the midgut epithelium in millipedes (Myriapoda,Diplopoda)

The midgut epithelia of the millipedes Polyxenus lagurus, Archispirostreptus gigas and Julus scandinavius were analyzed under light and transmission electron microscopies. In order to detect the proliferation of regenerative cells, labeling with BrdU and antibodies against phosphohistone H3 were employed. A tube-shaped midgut of three millipedes examined spreads along the entire length of the middle region of the body. The epithelium is composed of digestive, secretory and regenerative cells. The digestive cells are responsible for the accumulation of metals and the reserve material as well as the synthesis of substances, which are then secreted into the midgut lumen. The secretions are of three types – merocrine, apocrine and microapocrine. The oval or pear-like shaped secretory cells do not come into contact with the midgut lumen and represent the closed type of secretory cells. They possess many electron-dense granules (J. scandinavius) or electron-dense granules and electron-lucent vesicles (A. gigas, P. lagurus), which are accompanied by cisterns of the rough endoplasmic reticulum. The regenerative cells are distributed individually among the basal regions of the digestive cells. The proliferation and differentiation of regenerative cells into the digestive cells occurred in J. scandinavius and A. gigas, while these processes were not observed in P. lagurus. As a result of the mitotic division of regenerative cells, one of the newly formed cells fulfills the role of a regenerative cell, while the second one differentiates into a digestive cell. We concluded that regenerative cells play the role of unipotent midgut stem cells.     

Digestive enzymes of the saltmarsh periwinkleLittorina irrorata (Mollusca: Gastropoda)

Summary The saltmarsh periwinkleLittorina irrorata is well adapted for the digestion of a wide range of polysaccharides. Enzyme extracts attacked cellulose, pectin, xylan, bean gum and mannan (common in cell walls of higher plants), as well as starch and laminarin (representative of major polysaccharide classes in fungal, algal, and animal tissues). Activities were generally highes at a ph of 5 or 6. There was no evidence that chitin was digested, but 19 other enzymes, active toward various carbohydrates, lipids and peptides, were demonstrated. Enzymatic activity toward Azocoll, a general substrate for proteinase activity, was weak compared to that of other aquatic detritivores. The maximum was reached at a pH of 8. Enzymatic activities were generally measured with extracts of the entire visceral hump. Separate stomach or intestine extracts also gave strong activities. The stomach was the most acidic section of the digestive system with an average pH of 5.8; the intestine had an average pH of 7.3.     

Phylogeny of the Cocculinoidea (Mollusca, Gastropoda)

Abstract. The superfamily Cocculinoidea is a group of marine, deep-water, limpet-like gastropods. Recent speculation surrounding their affinities has concentrated on their placement within the Gastropoda. However, phylogenetic relationships within the Cocculinoidea, especially the monophyly of families and genera within the group, remain poorly understood. Phylogenetic analysis of 31 morphological characters for 15 cocculinoidean taxa and 2 outgroups resulted in a single most parsimonious tree, length=70, CI=0.62, and RI=0.71. Monophyly of the Cocculinoidea, Cocculinidae, and the genera Cocculina and Coccopigya was supported; Paracocculina and Coccocrater were found to be paraphyletic. Character optimization demonstrates that many characters often cited as diagnostic of various taxa, are often homoplastic and/or synapomorphies at different hierarchical levels.     

Secretory glands of the snail tentacle and their relation to the olfactory organ (Mollusca,Gastropoda)

Summary The epidermis of the posterior tentacles of the terrestrial snail Achatina fulica was examined by histological and histochemial methods. There are seven types of unicellular glands in the tentacle skin: three mucocytes containing either acid mucopolysaccharides or neutral mucopolysaccharides, or both; two mucocytes containing glycoproteins; a lipid gland; and a protein gland. The mucocytes are considerably more abundant along the shaft of the tentacle than at the tip, where the olfactory organ is situated. Conversely, the lipid glands and the protein glands are found almost exclusively in the olfactory organ. With minor exceptions, none of the foregoing cell types is present in the skin of the head or the foot. These observations indicate a high degree of local specificity in secretory products, consistent with a ubiquitous and generous endowment of glands in the molluscan skin. Collar cells, described by previous authors in closely related species, were not observed.     

Histochemical and ultrastructural studies on the salivary glands of Helix aspersa (Mollusca)

A histochemical and ultrastructural study was carried out on the salivary glands of adult specimens of Helix aspersa. Histochemically four cell types were distinguished: mucocyte I, mucocyte II, pseudochromosome cell and protein cell. The first secretes neutral and acid mucins. Mucocyte II and pseudochromosome cells produce acid mucins and the protein cell secretes neutral mucins and proteins.
The ultrastructural data for differentiating these four cell types is discussed. Mucocytes I have a characteristic arrangement of rough endoplasmatic reticulum and basal infoldings. Mucocytes II have pseudocrystalline formations in the rough endoplasmatic reticulum and intraluminary ducts. Pseudochromosome cells have typical granules with a reticular central zone. Protein cells have serous homogenous secretion.
The epithelium of the salivary ducts is described.     

Light and electron microscope investigations on the salivary glands of the slug,Agriolimax reticulatus (Müller)

Summary Ten cell types have been identified with the light microscope in the salivary glands ofA. reticulatus. Eight of these have also been observed with the electron microscope. A secretory cycle involving several cell types is absent. At least six different secretions are believed to be released from the glands. The pathways by which the secretions pass from the cells to the collecting ducts are outlined.Salivary duct epithelial cells have a characteristic ultrastructure which is similar to that found in ion and water transporting cells. It is proposed that the duct epithelial cells have a similar function.     

植物酶的组织和细胞化学定位研究方法

酶是参与植物体内生化反应的特殊蛋白质。在保持活组织和细胞结构完整性的条件下,利用组织化学、细胞化学、免疫学和显微检测等技术研究酶的即位定位,是了解酶在组织、细胞和亚细胞中的分布、活性动态与定量及酶功能等的重要途径。对植物体中酶定位的组织化学和细胞化学方法的概念、原理与研究进展进行了综述,并根据国际酶化学分类编号顺序,分别介绍了25种酶的组织化学染色定位所用的反应介质和染色方法及46种酶的细胞化学定位方法的参考文献。     

Rhipicephalus (Boophilus) microplus (Canestrini, 1887) (Acari: Ixodidae): acid phosphatase and ATPase activities localization in salivary glands of females during the feeding period

This study investigates the presence and the localization of acid phosphatase and ATPase in the salivary glands of Rhipicephalus (Boophilus) microplus female ticks during feeding. Semi-engorged females showed a larger amount of acid phosphatase compared to those at beginning of feeding, localized mainly in the apical portion of the secretory cells, and in the basal labyrinth of the interstitial cells. Ultrastructural observations also demonstrated its presence in secretion granules and inside some nuclei of secretory cells at beginning of feeding. Acid phosphatase in a free form probably has a hemolymph and/or ribosomal origin and participates in salivary gland secretion control. ATPase was detected in basal membrane of all types of acini and/or in the cytoplasm of the secretory cells at both feeding stages. The enzyme activities found strongly suggests that cell death by apoptosis occurs during the degenerative process.     

Reassessment of the trophic position of Bullidae (Gastropoda: Cephalaspidea) and the importance of diet in the evolution of cephalaspidean gastropods

A review of the available data on the diet of species of Bullidae is presented. Their trophic level is reassessed through gut content analysis of worldwide species from various habitats. Gut contents were identified by light and scanning electron microscopy. The previous claims of omnivory and carnivory were not supported. Worldwide species of Bullidae feed preferentially on algae and secondarily on diatoms, and are therefore herbivorous. The influence of dietary specialization on the evolution of cephalaspidean gastropods was assessed by superimposing the diet onto a molecular family-level phylogeny. Herbivory was found to be the plesiomorphic condition, with carnivory having arisen independently two or three times. This evidence suggests that dietary specialization has played a major role in the adaptive radiation of the group.