Localization and fate of aluminium in the digestive gland of the freshwater snail Lymnaea stagnalis

The digestive gland of the freshwater snail Lymnaea stagnalis, exposed to water containing an elevated concentration of aluminium at neutral pH for up to 30 days, followed by a 20 day recovery period, was examined by light and electron microscopy and X-ray microanalysis. Aluminium was localized in the yellow granules present in the digestive and excretory cells and in the green and small granules present in the digestive cells. More aluminium, silicon, phosphorus and sulphur were present in all three granule types from aluminium exposed snails. The number of yellow and green granules from the digestive gland of aluminium exposed snails showed a progressive increase over the experimental period compared to controls. The number and aluminium content of the granules is likely to reflect the role of the digestive gland as a 'sink' for accumulated aluminium. We propose that intracellular monomeric silica is involved in the detoxification of aqueous aluminium which at neutral pH is largely in the form of an insoluble polyhydroxide. The increased amounts of sulphur and phosphorus in the granules are likely to be part of a broad response to metal loading but probably do not play a significant role in the storage and detoxification of aluminium.     

ULTRASTRUCTURAL LOCALIZATION OF COMPLEX CARBOHYDRATES AND PHOSPHATASES IN DIGESTIVE GLAND CELLS OF FEEDING AND HIBERNATING SNAILS, HELIX LUCORUM (GASTROPODA: HELICIDAE)

The digestive gland of normally-fed snails Helix lucorum, aswell as that of snails which had hibernated for 4 months wereexamined by the use of cytochemical techniques for detectionof acid and alkaline phos-phatase, as well as of periodate-reactive(PA-TCH-SP technique), sulfated (HID-TCH-SP technique) and carboxylatedcarbohydrates (LID-TCH-SP technique). The cytochemical resultssupport the hypothesis of intracellular digestion via lysosomalactivity of material taken up by endocytotic processes by thecells of the digestive gland. Four months hibernation did notaffect the intracellular distribution of polysac-charides andphosphatases in the cells of the digestive gland of H. lucorumcompared to that in the control snails. In addition, hibernationaffected the percentage of the calcium cells which significantlyincreased compared to the non-hibernating snails, whiie thepercentages of the digestive and excretory cells remained almoststable. However, the periodate-reactive sulfated and carboxylatedpolysaccharides of the digestive gland cells decreased in thehibernated snails compared to the controls. The results suggestthat the cytochemistry of periodate-reactive, sulfated and carboxylatedpolysaccharides used in the present study could, also, be appliedto the study of lysosomal activities. (Received 1 October 1991; accepted 4 December 1991)     

The distribution of some hydrolytic enzymes in the cells of the digestive gland of certain lamellibranchs and gastropods

Five hydrolytic enzymes (acid phosphatase, aryl sulphatase, β-glucuronidase, N-acetyl-β-glucosaminidase, and non-specific esterase) have been studied histochemically in the cells of the digestive gland of Mytilus edulis, Helix aspersa , and certain other lamellibranchs and gastropods. All the enzymes studied have basically similar distributions.
In the digestive cells, the enzymes occur in cytoplasmic granules which are believed to be primary lysosomes; in vacuoles which contain phagocytosed food material; and in vacuoles containing lipofuscin granules, which are the residues of digestive activity.
In the basiphil cells of M. edulis , most of the enzymes are localized in a few cytoplasmic granules; non-specific esterase, however, is found throughout the cytoplasm. In the calcium cells of H. aspersa and the other pulmonate gastropods studied, the enzymes are either in cytoplasmic granules, or distributed diffusely throughout the cytoplasm. Acid phosphatase is also found in the calcium spherules, especially in H. aspersa.
In the excretory cells of H. aspersa and the other pulmonates studied, the enzymes are found in granules in the cytoplasm, and in the lipofuscin granules which lie in the vacuoles of these cells.     

Ultrastructural localization of esterase and acid phosphatase in digestive gland cells of fed and starvedCepaea nemoralis (L.) (Mollusca: Helicidae)

Summary The ultrastructural localizations of thiolacetic acid esterase, indoxyl acetate esterase and acid -glycerophosphatase have been studied in the digestive gland cells of fed and starvedCepaea nemoralis. In fed snails the major localization of all three enzymes was in the green granule vacuoles of digestive cells. In addition, the cytoplasm of calcium cells and the Golgi apparatus and GERL (?) of all cell types were acid phosphatase positive. Many digestive cells of starved snails showed a similar enzyme distribution to that found in fed snails but other digestive cells showed a very high cytoplasmic activity of all three enzymes. It is suggested that these cells are in the process of autolysis. New light is also thrown on the process by which food is transported from the digestive gland lumen to the phagosomes of digestive cells.     

The cytology,histochemistry, and ultrastructure of the cell types found in the digestive gland of the slug,Agriolimax reticulatus (Müller)

Summary Four cell types have been identified in the digestive glands from light and electron microscope studies. The possible functions of each cell type are discussed.Thin cells are undifferentiated. Calcium cells contain spherules of calcium salts which have a characteristic ultrastructure. Different protein granules are found apically. Digestive cells are present as two distinct forms. One form is believed to be absorbing food material and digesting it intracellularly, and the other form is a secreting cell. Both forms contain green and yellow granules and histochemistry shows these granules to be distinct. Protein granules also occur apically.Excretory cells are distinguished by having a large central vacuole containing excretory granules. Histochemistry shows these granules, like the yellow granules of digestive cells, to be composed mainly of lipofuscin.It is suggested that digestive cells form excretory cells.     

THE INFLUENCE OF DIETARY CALCIUM ON THE TISSUE DISTRIBUTION OF Cu, Zn, Mg & P AND HISTOLOGICAL CHANGES IN THE DIGESTIVE GLAND CELLS OF THE SNAIL ACHATINA FULICABOWDICHM

Dietary calcium had a varying effect on the concentration ofother metals located in the tissues and excreted in the faeces.Faecal phosphate, copper and zinc concentrations decreased withincreased dietary calcium but showed a peak of excretion 10–12weeks after the beginning of the experiment, while the totalamounts of the studied elements lost in the faeces increased.Phosphate values in the digestive gland of snails on calcium-supplementeddiets were lower than metal values found in control snails.Although the digestive gland magnesium concentrations were alwayslower and copper levels higher than the snails on the omnivorousdiet, the trend was for a decrease in tissue magnesium and copperwith increased calcium in the diet. In foot tissue, the trendwas towards a decrease in Mg with decreased dietary calcium.The zinc concentration in the digestive gland and foot of snailson the omnivorous diet was not different from the zinc levelsin snails on the three highest calcium-supplemented diets. Itis suggested that high levels of dietary calcium might act asan environmental stressor characterised by changes in the morphologyand histo-chemistry of the digestive tubules. (Received 14 August 1991; accepted 10 October 1991)     

Cellular cadmium distribution in the common winkle,Littorina littorea (L.) determined by X-ray microprobe analysis and histochemistry

Summary Various tissues of common winkles,Littorina littorea (L.), experimentally exposed to cadmium (Cd) chloride were examined using light and electron microscopy and their elemental composition determined by X-ray microanalysis and histochemistry. Membrane granules in gill epithelial cells with paddle cilia contain carbonates, phosphates and sulphides associated with different cations in different types of granules. Traces of Cd have been found only in those granules containing sulphur and iron. Nephrocytes also contain small amounts of this metal in the cytoplasm of excretory cells. X-ray microanalysis reveals that concretions of basophilic cells are minor sites for Cd sequestration while BTAN-ASSG stain for unbound Cd indicates that most of the Cd is located within the lysosomes of digestive cells in association with proteins. Low amounts of the metal have been evidenced in the granules of epithelial mantle cells rich in sulphur. The results also indicate that hemocytes contain granules of calcium phosphate and iron sulphide. Cd is also associated to sulphur rather than to phosphate. These hemocytes may act as Cd carrier from gills to kidney and digestive gland. A hypothetical pathway for Cd accumulation and detoxification is suggested.     

Cellular cadmium distribution in the common winkle, Littorina littorea (L.) determined by X-ray microprobe analysis and histochemistry

Various tissues of common winkles, Littorina littorea (L.), experimentally exposed to cadmium (Cd) chloride were examined using light and electron microscopy and their elemental composition determined by X-ray microanalysis and histochemistry. Membrane granules in gill epithelial cells with paddle cilia contain carbonates, phosphates and sulphides associated with different cations in different types of granules. Traces of Cd have been found only in those granules containing sulphur and iron. Nephrocytes also contain small amounts of this metal in the cytoplasm of excretory cells. X-ray microanalysis reveals that concretions of basophilic cells are minor sites for Cd sequestration while BTAN-ASSG stain for unbound Cd indicates that most of the Cd is located within the lysosomes of digestive cells in association with proteins. Low amounts of the metal have been evidenced in the granules of epithelial mantle cells rich in sulphur. The results also indicate that hemocytes contain granules of calcium phosphate and iron sulphide. Cd is also associated to sulphur rather than to phosphate. These hemocytes may act as Cd carrier from gills to kidney and digestive gland. A hypothetical pathway for Cd accumulation and detoxification is suggested.     

Localization of Metals in the Gastropod Littorina saxatilis (Prosobranchia: Littorinoidea) from a Polluted Site

Organ heavy metal levels, and ultrastructural localization were examined in the marine prosobranch mollusc Littorina saxatilis from a metal-contaminated site. Copper and zinc are localized in the digestive gland, stomach and kidney. The digestive gland does not appear to be a significant site of iron accumulation. High levels of copper and zinc in the stomach may indicate significant uptake from the food. Relatively high heavy metal levels in several organs were reflected in the presence of a variety of metal-containing aggregates (granules) within the tissues. In common with other invertebrates two broad classes of granule were present. The first appears to be proteinaceous, with a high sulphur content. This type contained copper and iron and was found in the stomach, kidney, pore cells, and rectum. The second type was inorganic and mineralized, containing phosphorus. They contained calcium, zinc and manganese, and were observed in the kidney and digestive gland. Differences in the number of metals were observed between the various granule types, possibly reflecting specific pathways involved in metal sequestration.     

Histopathological effects of larval digenea on the digestive epithelia of the marine prosobranch Cerithidea californica: Fine structural changes in the digestive gland

The effects of natural infections by rediae of the heterophyid Euhaplorchis californiensis on the digestive gland cells of Cerithidea californica (Gastropoda: Prosobranchiata) were studied through the use of light and electron microscopy. Pathological changes in secretory and digestive cells, including the disruption of digestive vacuoles and cell-cell junctions, the deformation of lumenal microvilli, the lysis of distal plasma membranes, and nuclear polymorphism, usually accompany the migration of larval trematodes into the digestive gland. Degenerating secretory cells are further characterized by Golgi complexes which assume an inflated appearance and a reduction of rough endoplasmic reticulum in the cell's perinuclear region. Finally, an increase in excretory and autophagic activities in the digestive gland cells of infected snails may be considered possible mechanisms by which the host is able to compensate for adverse metabolic or nutritional stresses of trematode parasitism.     

HISTOCHEMISTRY AND ULTRASTRUCTURE OF THE CRYPT CELLS IN THE DIGESTIVE GLAND OF APLYSIA PUNCTATA (CUVIER, 1803)

The crypt cells lining the Aplysia punctata digestive tubulescomprise of three types of cell; calcium, excretory, and thincells. The calcium cells play a role in osmoregulation, mineral storage,exocrine secretion, iron detoxification, and excretion processes.They possess well- developed microvilli and a basal labyrinth,suggesting a role in absorption. The Golgi apparatus is involvedin the production of two main components of calcium spherules;the fibrillar material and mineralized granules. Golgi complex,rough endoplasmic reticulum (RER), ribosomes, and altered mitochondriaare involved in the formation of calcium spherules. Secretoryactivity is indicated by the formation of dense granules containingiron and calcium salts. Lipofuscin pigment has been found inlarge concretions which may arise from cytoplasmic areas surrounded byendoplasmic reticulum, RER and Golgi tubules. There are threestages of excretory cells, called early, mature, and post-excretorycells. This study traces the development of granulofibrillarvacuoles up to the formation of the lipofuscin concretions andshows that excretory cells are in fact degenerating calciumcells. The fine structure of thin cells suggests that they areyoung calcium cells. (Received 29 December 1997; accepted 15 November 1998)     

铜锈环棱螺肝脏的显微与超微结构观察

应用组织切片、光学显微镜及透射电子显微镜技术,对铜锈环棱螺肝脏进行观察研究。结果表明,铜锈环棱螺肝脏为复管泡状腺,由许多肝小叶组成,每个肝小叶管壁由单层上皮细胞构成,分3种细胞：消化细胞、排泄细胞、钙细胞。光镜显示,肝小叶多球形或近球形,横切面管腔呈星射状裂隙,纵切面可见上皮细胞呈柱状,核靠近基膜,上皮细胞极性明显。消化细胞构成腺上皮的主体,染色较淡。排泄细胞和钙细胞染色较深。肝腺管之间有不规则圆形的网状细胞,结缔组织填充在肝小叶之间及小叶与导管之间。电镜显示,消化细胞和排泄细胞顶端都具微绒毛,消化细胞主要特征为具有溶酶体囊泡系统,参与细胞内消化过程。排泄细胞中粗面内质网、线粒体等细胞器含量丰富,细胞内还散布着大量脂滴。钙细胞锥体形,主要特征为细胞内具钙粒子,也含有大量细胞器。     

Mineral congregations, “spherites” in the midgut gland ofCoelotes terrestris (Araneae): Structure,composition and function

Summary Spherites in the digestive and secretory cells of the midgut gland of the agelenid spiderCoelotes terrestris were studied by electron microscopy and histochemical methods. Spherites measured 1–6 m in diameter and were characterized by alternating layers of electron dense and electron lucent material. The main-components of spherites were calcium phosphates and calcium carbonates. Guanine and barium, as well as aminopeptidase and alkaline phosphatase were also present. The matrix consisted of proteins and carbohydrates. Numerous spherites were found together with excretory products within the excretory vacuoles of the digestive cells.Spiders fed with food containing lead, showed deposition of the metall in the spherites. It is then proposed that spherites, aside from their role in storing calcium and other ions, may function in detoxification of heavy metals.     

Histochemical changes of carbohydrate and protein contents in the digestive gland cells of the land snail Monacha cartusiana following starvation

The present study was designed to investigate histochemically the detection of carbohydrate and protein in the normally feeding snails and after 15 and 30 days of starvation. Generally, abundant carbohydrate and protein materials were detected in the component cells of the digestive gland of normally feeding snails. The results of this investigation revealed a pronounced decline of carbohydrates in the digestive gland cells of Monacha cartusiana snails after starvation. Severe decline in carbohydrate content was observed especially after 30 days of starvation. Moreover, protein inclusions have exhibited a week stainability in the digestive gland cells of these snails as a consequence of starvation.     

Calcium Dynamics in Land Gastropods

In land gastropods, calcium is precipitated in the shell, inconnective calcium cells which are largely distributed throughthe whole connective tissue, in epithelial calcium cells ofthe digestive gland, and in the calcium gland cells of the skinand the mantle collar. Calcium is taken up from the externalmedium by food and by absorption through the sole skin. To adaptto terrestrial life, these animals have to eliminate appreciableamounts of calcium for their protection and their reproduction.During the egg laying period, a calcium flux occurs throughthe epithelium of the reproductive tract in order to supplythe egg shell and the egg fluids. This egg calcium is takenup by the embryo. The maintenance of a positive calcium balancebetween its uptake and the loss is due to an important reservoirof easily mobilizable calcium in the form of calcium carbonate.This reservoir consists of the connective calcium cells whichare constantly able to accumulate or release calcium as longas calcium is locally available or required. The epithelialcalcium cells of the digestive gland are loaded with calciumphosphate; they are not a major calcium storage compartment,but have an essential function in detoxification. All of thecalcium movement occurring across cell membranes and throughepithelia concerns only calcium ions. All calcium movement canbe regarded either as on-off systems or as reversible systems,both of which are certainly controlled by complex processes     

Effect of chronic and acute lead treatment in the slug Arion ater on calcium and delta-aminolaevulinic acid dehydratase activity

Accumulation of lead in the tissues was greater after acute exposure. Most of the metal was deposited in the intestine and least in foot. After chronic exposure to lead the calcium content of the intestine decreased. After chronic and acute exposure to lead the calcium content of the digestive gland and foot was lower than control values. Lead in the digestive gland granules was higher after acute lead treatment. Less than 45% of the lead was associated with the granules. These results are discussed in view of the difference in metabolism and chemical nature of the organs investigated. ALAD activity was located in the digestive gland and it was reduced after acute treatment with lead.     

DISTRIBUTION OF METALS IN THE DIGESTIVE GLAND -- GONAD COMPLEX OF THE MARINE GASTROPOD NUCELLA LAPILLUS

The amounts of phosphate, zinc, copper and calcium in gonadmaterial from Nucella lapillus (L.) was higher in males comparedwith females. Water insoluble extracts from the digestive glandwere found to contain granules high in concentrations of phosphate,magnesium, zinc, copper and calcium when compared with watersoluble extracts and both types of extract from gonad tissueof male and female dog whelks. Male N. lapillus contained thehighest amounts of zinc and copper in isolated digestive glandgranules. Intracellular granules containing metal were locatedhistochemically in cells of the digestive gland and visceralhaemocoelic spaces (Received 5 December 1978;     

Phosphatase activity in the hepatopancreas of Helix aspersa

• 1.1. Phosphatase acid (PhA) activity in the digestive gland (hepatopancreas) of the common garden snail Helix aspersa has been investigated using cytochemical methods.
• 2.2. All the cells composing this gland show PhA activity, the distribution pattern differing according to the cell type.
• 3.3. The digestive cells show the most widely distributed reaction product (brush border, phagolysosomes, multivesicular bodies and autophagic vacuoles).
• 4.4. In the excretory cells this activity appears in large sacs, while in the calcium cells the reaction product is abundant in the calcium granules.
• 5.5. Cellular digestion processes performed by each of these cell types is discussed together with their role in the detoxification of heavy elements derived from the environment.

     

The ultrastructural investigation of the midgut in the quill mite Syringophilopsis fringilla (Acari,Trombidiformes: Syringophilidae)

The midgut of the females of Syringophilopsis fringilla (Fritsch) composed of anterior midgut and excretory organ (=posterior midgut) was investigated by means of light and transmission electron microscopy. The anterior midgut includes the ventriculus and two pairs of midgut caeca. These organs are lined by a similar epithelium except for the region adjacent to the coxal glands. Four cell subtypes were distinguished in the epithelium of the anterior midgut. All of them evidently represent physiological states of a single cell type. The digestive cells are most abundant. These cells are rich in rough endoplasmic reticulum and participate both in secretion and intracellular digestion. They form macropinocytotic vesicles in the apical region and a lot of secondary lysosomes in the central cytoplasm. After accumulating various residual bodies and spherites, the digestive cells transform into the excretory cells. The latter can be either extruded into the gut lumen or bud off their apical region and enter a new digestive cycle. The secretory cells were not found in all specimens examined. They are characterized by the presence of dense membrane-bounded granules, 2–4 μm in diameter, as well as by an extensive rough endoplasmic reticulum and Golgi bodies. The ventricular wall adjacent to the coxal glands demonstrates features of transporting epithelia. The cells are characterized by irregularly branched apical processes and a high concentration of mitochondria. The main function of the excretory organ (posterior midgut) is the elimination of nitrogenous waste. Formation of guanine-containing granules in the cytoplasm of the epithelial cells was shown to be associated with Golgi activity. The excretory granules are released into the gut lumen by means of eccrine or apocrine secretion. Evacuation of the fecal masses occurs periodically. Mitotic figures have been observed occasionally in the epithelial cells of the anterior midgut.     

Giant secretory granules in the ducts of the parotid and submandibular glands of the slow loris

The parotid and submandibular glands of a slow loris, a rare Southeast Asian primate, were obtained after the head had been perfused by fixative for a study of the brain. These tissues were processed by conventional means for electron microscopy. Glands also were obtained at autopsy from 2 other lorises, fixed by immersion in formalin, and subjected to a battery of tests for glycoconjugates. In the parotid gland, a short segment of the proximal striated duct lacks both basal striations and any sign of secretory activity. The major portion of the striated duct consists of tall cells that contain a spectrum of secretory granules, some larger than the nuclei (many granules are > 9 mum in diameter). These granules, which are delimited by a single membrane, are capable of chain exocytosis. Many of the giant granules have bundles of cytofilaments (4.5-6.5 nm) in apparent association with their surface. Occasional cells contain numerous small granules. Duct cells with or without granules lack basal striations. The granules contain neutral glycoconjugates but no acidic glycoconjugates. Some, but not all, interlobular excretory ducts also have secretory granules that run the gamut from tiny to giant. Exactly the same situation occurs in the submandibular gland. Unlike other primates, which may have duct cells that contain only a few tiny granules in their apices, the cells in both the striated and excretory ducts in the slow loris appear to be specialized for secretion rather than for transport. The biofunction of the giant granules is unknown.