Fine structure of the axillary gland in the brown bullhead (Ictalurus nebulosus)

The axillary glands of Ictalurus are lobulated invaginations of the epidermis, opening at a pore between the pectoral spine and the cleithrum. Holocrine cells lining a false lumen form a viscous secretion. The secretory cells originate in the tenuous basal layer of the gland wall. Secretion is initiated by the formation of compound vesicles in cells that become very large and have complex cytoplasm of a varied appearance. Golgi systems are well developed and the perinuclear cytoplasm may contain many mitochondria and sacs of ribosomal endoplasmic reticulum; some tracts of cytoplasm are vesicular and contain free ribosomes. Some cells contain numerous large lysosomes, and some have extensive contents of fibrillar masses imperfectly separated by membranes, that recall the appearance of the mucous secretion of goblet cells. The secretory cells break down, releasing the degenerating organelles, including the nuclei, into the false lumen. Some structures are still recognizable in the secretion even after it has been expelled, but the main part of the formed secretion consists of the mucus-like masses. Various leucocytes are found in the gland walls and embedded in the secretion. The fine structure differentiates the holocrine cells of the axillary gland from the club cells of the epidermis, and from the venom glands associated with the fin spines of catfishes. The function of the axillary gland secretion remains unknown.     

Ultrastructural observations on the erythrocytes and thrombocytes of the tuatara, Sphenodon punctatus (gray)

The erythrocytes of Sphenodon punctatus (Gray) are nucleated, ellipsoidal and flattened, and contain 55--65 microtubules in their marginal band. The thrombocytes are also flattened, ellipsoidal, nucleated cells and in electron-microscopic preparations occurred in aggregrates. The thrombocytes appeared to be 'activated' and possessed many pseudopodia which were devoid of organelles. The latter were concentrated in the perinuclear region and were encircled by a ring of microtubules. The organelles included ribosomes, mitochondria, membrane--bound dense material and numerous actin-like microfilaments. Cytoplasmic vacuoles contained a moderately dense, filamentous material and/or spheroidal electron-dense inclusions, beta-glycogen particles were scattered in the general cytoplasm and were most concentrated in the pseudopodia. The erythrocytes and thrombocytes of S. punctatus are compared with those in other vertebrates.     

An ultrastructural investigation of the early stages of oocyte differentiation in Actinia fragacea (Cnidaria; Anthozoa)

Individuals from a population of the intertidal sea anemone Actinia fragacea (Tugwell) were collected at approximately monthly intervals over an 18 month period. Samples of gonad were removed from each anemone and examined by light and electron microscopy. During late spring and early summer, large numbers of small cells were seen in the endoderm of the female gonads, lying close to the mesoglea. For convenience, these cells were classified into three types. Type I cells are 6–9 μm in diameter, with relatively very large nuclei, which may contain synaptinemal complexes, and scant cytoplasm containing few organelles. Type II cells are larger, reaching 15 μ m in diameter, with more abundant cytoplasm containing more organelles and inclusions. The nucleus is more dense, but may also contain synaptinemal complexes. Type III cells are less common. They are similar in size to Type II cells, but their nuclei contain irregular dense chromatin masses, and the nuclear envelope is incomplete or absent. The possible significance of the various cell types is discussed. It is suggested that Type I cells are oocytes at a very early stage of differentiation and that Type II cells are rather later oocytes. The status of the Type III cells is uncertain.     

Developmental stages and localization of peroxidatic activity in the leucocytes of three teleost species (Cyprinus carpio L.; Tinca tinca L.; Salmo gairdneri Richardson)

Summary The ultrastructural localization of peroxidase (PO) in the leucocytes of three teleosts (Cyprinus carpio L., Tinca tinca L., Salmo gairdneri R.) has been investigated using the 3,3-diaminobenzidine method. In the heterophilic granulocytes the granules show a species specific structure and are PO-positive at pH 7.6. They can be traced back to small granules arising near the Golgi apparatus (GA) in the promyelocyte. They coalesce to form larger granules and gradually change into the mature type. Myelocytes contain small unreactive granules, and these represent a second granule population. Eosinophils contain one PO-positive granule type (at pH 9), and these granules show a varying density during cell maturation.Basophils are present only in the Cyprinid species, and contain unreactive granules originating from precursors displaying a weakly positive reaction at pH 7.6. The active secretory organelles (RER, GA) are PO-negative, except for a weakly positive reaction in the flocculent matrix of the inner G-cisternae.In promonocytes and monocytes the granules are unreactive, but in the macrophages PO-positive staining occurs in a few small to medium sized granules, and in large vacuoles. At least some of these latter are apparently derived from phagolysosomes containing digested erythrocytes. Thrombocytes and lymphocytes are unreactive.The successive development of PO-positive and negative granule populations in the heterophils, and the PO-reactivity of eosinophils and basophils, show some similarities to the corresponding cells in higher vertebrates, but an analogous PO-positive (azurophil) granule type in monocytes seems to be absent.     

Role of adenylate kinase, AMP deaminase and 5'-nucleotidase in the metabolism of adenylic nucleotides

The activities of adenylate kinase, AMP-deaminase and 5'-nucleotidase in various tissues of the rat were studied. The activity of the forward adenylate kinase reaction (ATP + AMP----2 ADP) against the back one (2 ADP----ATP + AMP) was predominant. The liver was shown to contain two, while the blood serum--three adenylate kinase isoenzymes. In the skeletal muscles, the catabolism of adenylic acid involving AMP-deaminase and 5'-nucleotidase predominantly occurred via deamination, in the liver--via dephosphorylation, while in the leucocytes, erythrocytes and blood serum the activity of these processes was essentially the same. In vitro, ATP enhanced the activity of AMP-deaminase in the liver, leucocytes and erythrocytes and decreased it in the blood serum. Under effects of ATP, the activity of 5'-nucleotidase in the leucocytes and blood serum was markedly elevated, that in the liver and erythrocytes was unaffected.     

Fine Structural Observations of the Erythrocytic Stages of Plasmodium chabaudi Landau, 1965*

SYNOPSIS. Electron microscopic examination of Plasmodium chabaudi in mouse erythrocytes revealed many characteristics resembling those observed in other mammalian malarial parasites. A double unit membrane surrounds the trophozoite cytoplasm which contains many ribonucleoprotein particles, a limited amount of endoplasmic reticulum and membraned organelles including sausage-shaped vacuoles and multilaminated membraned bodies. More or less circular double membraned vacuoles, possibly cross sections of the sausage-shaped vacuoles, are common. Typical protozoan mitochondria are lacking. The limiting membrane of the merozoites is triple-layered. Paired organelles and small dense bodies are found in the merozoites along with dense granular masses in the nuclei. Trophozoites have cytostomal structures as well as invaginations of the plasma membrane at sites where no cytostomes are evident. Digestion appears to occur in single membrane-bound vesicles which contain one to several pigment grains. P. chabaudi frequently contains multiple food vacuoles and has polymorphism manifested in part by the presence of cytoplasmic extensions and of nuclei with a variety of shapes. Several apparently free forms are noted, often accompanied by a thin rim of host cytoplasm. “Appliqué” forms are common among the trophozoites as are forms in which 2 or more trophozoites are joined together. Finally, alterations in the host cytoplasm resembling the socalled Maurer's clefts are frequent. Ferritin-containing vacuoles also appear in the host cell.     

Ultrastructural observations on the gallbladder of Richardson's ground squirrel (Spermophilus richardsonii)

The gallbladder of Richardson's ground squirrel shares many structural similarities with numerous other species, including the mouse, rabbit and man. Dark cells exhibit a condensed cytoplasm but contain all the typical organelles of light cells, although their total complement appears reduced. Edematous cells extruded from the epithelial lining were replaced through mitosis or had their space occupied by adjacent cells. Specializations of the basal plasma membrane and underlying basal lamina form peg-and-socket interdigitations, and desmosomes were observed on the lateral cell boundaries down to the level of the basal lamina. The latter remain intact during active water resorption while the former disappear. Adrenergic nerve fibers were observed intramuscularly and beneath the basal lamina. Random interepithelial migration of leucocytes was also observed. Some of these observations help to elucidate controversial issues whereas others have been previously unreported and may be species-dependent.     

LIGHT AND ELECTRON MICROSCOPY OF A PUNCTATE SPECIES OF PYRAMIMONAS,P. MUCIFERA SP. NOV. (PRASINOPHYCEAE)1

Pyramimonas mucifera sp. nov., a punctate species of the genus, is unusual both behaviorally and at the fine structural level. It forms two distinct populations in culture, one benthic and one planktonic. Planktonic forms are more conventional for the genus, but benthic forms are found in loosely packed mucilage, have flagellar rather than ciliary beating of the flagella, and display a higher degree of metaboly. Ultrastructurally this species is unusual in that it has a unique scale complement and the cells contain numerous muciferous vesicles, leaving only small pockets of cytoplasm containing the usual organelles. This species has a 3–1 type flagellar apparatus but has an additional fibrillar band, a 4–3-2–3 microtubular root system and a flexible synistosome. The discovery of a mucilage-producing species of Pyramimonas draws attention to possible links with other prasinophytes (Halosphaera) and green algae of questionable affiliation (Oltmannsiellopsis and Hafniomonas). It also provides a model of the primitive pyramimonad.     

The spermatogenous body cell of the coniferPicea abies (Norway spruce) contains actin microfilaments

Summary In conifer pollen, the generative cell divides into a sterile stalk cell and a body cell, which subsequently divides to produce two sperm. InPicea abies (Norway spruce, Pinaceae) this spermatogenous body cell contains actin microfilaments. Microfilament bundles follow the spherical contour of the body cell within the cell cortex, and also traverse the cytoplasm and enmesh amyloplasts and other organelles. In addition, microfilaments are associated with the surface of the body cell nucleus. The sterile stalk cell also contains microfilament bundles in the cytoplasm, around organelles, and along the nuclear surface. Within the pollen grain, microfilament bundles traverse the vegetative-cell cytoplasm and are enriched in a webbed cage which surrounds the body cell. Microfilaments were identified with rhodamine-phalloidin and with indirect immunofluo-rescence using a monoclonal antibody to actin. The majority of evidence in literature suggests that the spermatogenous generative cell in angiosperms does not contain actin microfilaments, so the presence of microfilaments within the spermatogenous body cell inP. abies appears to be a fundamental difference in sexual reproduction between conifers and angiosperms.     

FROM PROTOPLASM TO SWARMER: REGENERATION OF PROTOPLASTS FROM DISINTEGRATED CELLS OF THE MULTICELLULAR MARINE GREEN ALGA MICRODICTYON UMBILICATUM (CHLOROPHYTA)1

Protoplast regeneration from extruded cytoplasm of the multicellular marine green alga Microdictyon umbilicatum (Velley) Zanardini (Cladophorales, Anadyomenaceae) was investigated. The early process of protoplast formation is comprised of two steps: agglutination of cell organelles into protoplasmic masses followed by generation of a temporary enclosing envelope around them. Agglutination of cell organelles was mediated by a lectin–carbohydrate complementary system. Three sugars, D‐galactosamine, D‐glucosamine, and α‐D‐mannose, inhibited the agglutination process, and three complementary lectins for the above sugars, peanut agglutinin, Ricinus communis agglutinin, and concanavalin A, bound to the surfaces of chloroplasts. Agglutination assay using human erythrocytes showed the presence of lectins specific for the above sugars in the algal vacuolar sap. A fluorescent probe 1‐(4‐trimethylammoniumphenyl)‐6‐phenyl‐a, 3,5‐hexatriene revealed that the envelope initially surrounding protoplasts was not a lipid‐based cell membrane. However, this developed several hours later. Simultaneous fluorescein diacetate and propidium iodide staining showed that the primary envelope had some characteristics of cell membranes, such as semipermeability and selective transport of materials. Also, fluorescein diacetate staining showed esterase activity in the protoplast and relocation of cell organelles and compartmentalization of cytoplasm during the process of regeneration. Both pH 7–9 and salinity 400–500 mM were found to be essentially important for the development of the protoplast envelope. When the basic regeneration process was accomplished, two alternative pathways of development were seen; about 70% of one‐celled protoplasts transformed into reproductive cells within 2 weeks after wounding, whereas others began cell division and grew into typical Microdictyon thalli. Quadriflagellate swarmers were liberated from the reproductive cells, and they germinated into mature individuals. It is therefore suggested that this species may use the wound response as a method of propagation and dispersal.     

Ultrastructure of Microvillar Coelomocytes from the Trunk Coelom of Thysanocardia nigra Ikeda, 1904 (Sipuncula) from the Sea of Japan

This research is part of a study on the ultrastructure of coelomocytes and cellular complexes from the body cavity of sipunculans. New free-swimming elements called microvillar cells in the trunk coelom of Thysanocardia nigra Ikeda, 1904 are examined using transmission electron microscopy. The cell harbors a giant vesicle filled with a fibrous matrix and rosettes of minute osmiophilous granules. The nucleus is peripheral, and a few cell organelles are situated between the cell membrane and the vesicular membrane. The cell membrane bears numerous microvilli with enlarged apical points. Numerous small microvillar vesicles swimming in the coelomic fluid separate from the microvillar cells. The functional morphology of coelomocytes and cellular complexes is discussed.Original Russian Text Copyright © 2005 by Biologiya Morya, Maiorova, Adrianov.     

Two New Species of Nosema (Microsporida: Nosematidae) from the Mexican Bean Beetle Epilachna varivestis (Coleoptera: Coccinellidae)1,2

Two new species of the genus Nosema (Microsporida: Nosematidae) are described from the Mexican bean beetle, Epilachna varivestis (Coleoptera: Coccinellidae) and their life cycle stages studied by light and electron microscopy. Both species are monomorphic and disporous: they develop in direct contact with the cytoplasm of host cells and the nuclei of all stages are diplokaryotic. The more virulent species produces systemic infections most extensively in the adipose tissue, muscles, and Malpighian tubules of larvae and also invades the reproductive tissues of adult beetles. During merogonic development, it forms chains of diplokaryotic meronts. The fine structure of the sporoblast nuclei shows clumped material in the pole of each nucleus opposite their common plane of apposition. Spores are straight to slightly curved and ovocylindrical in shape and they measure 5.3 ± 0.13 × 2.1 ± 0.03 μm. The less virulent species also invades most host tissues but does not develop in the midgut epithelium; the Malpighian tubules are the principal site of its development and it also invades the ovaries and testes of adult beetles. Merogony occurs exclusively as the result of binary fission of diplokaryotic meronts. The plasmalemma of the meronts is covered with a thin deposit of exospore material upon which are located closely packed tubules that encircle the body transversely. A thickened deposit of exospore material on the surface of the diplokaryotic sporonts later obscures these tubules. Other tubules occur free in the host cell cytoplasm or attached to the plasmalemma of meronts and sporonts. Secretory granules also occur free or in chains in the host cytoplasm and are probably produced from the surface of the sporoblasts. Sporoblasts also contain an unusual cup-shaped organelle associated with a dense body, which is apparently involved in the formation of the polar tube and its associated organelles in the anterior part of the spore. Spores are ellipsoidal to slightly pyriform and measure 4.7 ± 0.06 × 2.6 ± 0.03 μm.     

Some Corrections in Haemogregarine (Apicomplexa: Protozoa) Nomenclature1

The nomenclature of three genera in the family Haemogregarinidae (Haemogregarina, Karyolysus, and Hepatozoon) has been reviewed and the following new names are introduced to replace homonyms or for previously unnamed species: Haemogregarina carlosi n. nom., in the erythrocytes of the lizard Lacerta ocellata; Haemogregarina tincae n. nom., in the stomach and intestine of the tench Tinca tinca; Hepatozoon insectivorae n. sp., in the leucocytes of the shrews Sorex araneus and Crocidura leucodon; Hepatozoon krampitzi n. sp., in the leucocytes of the vole Microtus oeconomus; Hepatozoon peromysci n. sp., in the leucocytes of the deermice Peromyscus boylii and P. truei gilberti; and Hepatozoon pallida (Pessoa et al., 1971) n. comb., in the erythrocytes of the snake Thamnodynastes pallidus nattereri.     

PURIFICATION AND CHARACTERIZATION OF A LECTIN,BRYOHEALIN, INVOLVED IN THE PROTOPLAST FORMATION OF A MARINE GREEN ALGA BRYOPSIS PLUMOSA (CHLOROPHYTA) 1

When the coenocytic green alga Bryopsis plumosa (Huds.) Ag. was cut open and the cell contents were expelled, the cell organelles agglutinated rapidly in seawater to form protoplasts. Aggregation of cell organelles in seawater was mediated by a lectin–carbohydrate complementary system. Two sugars, N‐acetyl‐d ‐glucosamine and N‐acetyl‐d ‐galactosamine inhibited aggregation of cell organelles. The presence of these sugars on the surface of chloroplasts was verified with their complementary fluorescein isothiacyanate‐labeled lectins. An agglutination assay using human erythrocytes showed the presence of lectins specific for N‐acetyl‐d ‐galactosamine and N‐acetyl‐d ‐glucosamine in the crude extract. One‐step column purification using N‐acetyl‐d ‐glucosamine‐agarose affinity chromatography yielded a homogeneous protein. The protein agglutinated the cell organelles of B. plumosa, and its agglutinating activity was inhibited by the above sugars. Sodium dodecyl sulfate polyacrylamide gel electrophoresis results showed that this protein might be composed of two identical subunits cross‐linked by two disulfide bridges. Enzyme and chemical deglycosylation experiments showed that this protein is deficient in glycosylation. The molecular weight was determined as 53.8 kDa by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry. The N‐terminal 15 amino acid sequence of the lectin was Ser–Asp–Leu–Pro–Thr–X–Asp–Phe–Phe–His–Ile–Pro–Glu–Arg–Tyr, and showed no sequence homology to those of other reported proteins. These results suggest that this lectin belongs to a new class of lectins. We named this novel lectin from B. plumosa“bryohealin.”     

ULTRASTRUCTURE OF THE CORALLINACEAE (RHODOPHYTA) II. VEGETATIVE CELLS OF LITHOTHRIX ASPERGILLUM1

The ultrastructure of the calcareous red coralline alga Lithothrix aspergillum Gray and the development of the various tissue types has been studied. The sub-apical meristematic tissue alternately produces genicular or intergenicular cells. The genicular cells rapidly elongate and their cell walls thicken and become denser as more fibrillar wall material is laid down within the cell wall. These cells contain little cytoplasm and few organelles. The inter genicular cells which elongate only slightly during development have a small vacuole and many free starch grains in the cytoplasm. The peripheral cells in each inter genicular layer remain meristematic and form a cortical cell layer over the genicular cells. These cortical cells and the apical meristematic cells are covered by small epidermal cells which have extensive cell wall ingrowths between the chloroplasts. The inter genicular cells are calcified. Although the CaCO₃ is laid down within the cell walls, there is always a thin layer of CaCO₃-free organic cell wall material between the plasmalemma and the CaCO₃ impregnated wall. Only the distal tips of the genicular cells are calcified. In old genicular tissues of Lithothrix, secondary deposits of CaCO₃ of unknown crystallography are also found in the spaces between the cell walls. Thus there appear to be at least two mechanisms of calcification in this alga.     

Ultrastructure of Coelomocytes in Sphaerolaimus gracilis de Man, 1876 (Nematoda)

The structure of coelomocytes in the adenophorean aquatic nematode Sphaerolaimus gracilis de Man 1876 was studied with light and electron microscopes. Acid phosphatase and catalase activities were demonstrated by electron microscopy. Two pairs of coelomocytes occurred laterally posterior to the esophagointestinaljunction. The anterior pair of the coelomocytes, with the renette cell and gonad, lay in either the left or the right lateral side of the body. The posterior pair of coelomocytes was in the opposite side of the body, usually posterior to the renette. A long, thin, cell-extension-like structure appeared to originate from the coelomocytes. Coelomocytes were characterized by specialized organelles (CC-organelle) and large vacuoles. The CC-organelle contained crystalline structures like those in peroxisomes. Acid phosphatase and catalase activities were detected in the matrix of CC-organelles and catalase activity in the vacuoles. It was assumed that vacuoles originate from the CC-organelles. Coelomocytes showed pinocytotic activities, and numerous vesicles were observed between the cell membranes and the vacuoles.     

Leaf glands act as nectaries in Diplopterys pubipetala (Malpighiaceae)

Leaf glands of Diplopterys pubipetala were studied with light and electron microscopy. Aspects of their secretion, visitors and phenology were also recorded. Glands occur along the margin, at the apex and at the base of the leaf blade. All the glands begin secretion when the leaf is still very young, and secretion continues during leaf expansion. The highest proportion of young leaves coincides with the beginning of flowering. The glucose‐rich secretion is collected by Camponotus ants, which patrol the newly formed vegetative and reproductive branches. All the glands are sessile, partially set into the mesophyll, and present uniseriate epidermis subtended by nonvascularised parenchyma. The glands at the apex and base are larger and also consist of vascularised subjacent parenchyma. The cytoplasm of epidermal and parenchyma cells has abundant mitochondria, polymorphic plastids filled with oil droplets and a few starch grains. Golgi bodies and endoplasmic reticulum are more abundant in the epidermal cells. The parenchyma cells of the subjacent region contain chloroplasts and large vacuoles. Plasmodesmata connect all the nectary cells. The zinc iodide–osmium tetroxide (ZIO) method revealed differences in the population of organelles between epidermal cells, as well as between epidermal cells and parenchyma cells. Ultrastructural results indicate that leaf glands of D. pubipetala can be classified as mixed secretory glands. However, the secretion released by these glands is basically hydrophilic and composed primarily of sugars, hence these glands function as nectaries.     

Ultrastructural localization of RNA in the chromatoid bodies of undifferentiated cells (neoblasts) in planarians by the RNase–gold complex technique

Undifferentiated cells of planarians (Platyhelminthes, Turbellaria), also called neoblasts, are totipotent stem cells, which give rise to all differentiated cell types, while maintaining their own density by cell proliferation. Neoblasts are the only somatic cells of planarians bearing chromatoid bodies in their cytoplasm; these organelles disappear as differentiation takes place. Studies on germinal cells of several groups of organisms have shown that chromatoid bodies contain substantial amounts of RNA. To test its presence in neoblasts, we have used an RNase–gold technique. We found chromatoid bodies labeled with RNase–gold particles. Heterogeneity in the density of the label, may be correlated with the functionality and complexity of these organelles. The gold marker was also present over the nucleus and rough endoplasmic reticulum, but mitochondria, secretory granules, and the extracellular space were devoid of label. This specific localization of RNA in planarian chromatoid bodies supports earlier findings on germ cells and embryonic cells in a variety of organisms, indicating that chromatoid bodies are information-storage structures, essential during the process of cell differentiation. © 1993 Wiley-Liss, Inc.     

Cellular Responses of Congenital Immunity in the Starfish <Emphasis Type="Italic">Asterias rubens</Emphasis>

The work deals with analysis of changes of cellular defense factors in the starfish Asterias rubens in response to injection of human erythrocytes (HE). The number of circulating coelomocytes, dynamics of their production of active oxygen forms, activity of peroxidase, and dynamics of elimination of human hemoglobin from coelomic fluid were estimated before immunization with HE as well as at 6–144 h. The number of coelomocytes was counted in Goryaev chamber, production of active oxygen forms was determined in the test of spontaneous and zymosan-induced reduction of Tetrazolium Nitro Blue, peroxidase activity—in a color enzymatic reaction. Time of human hemoglobin elimination from the coelomic fluid was determined by spectrophotometric method by hemoglobin binding with acetone cyanohydrin with formation of a colored product. It is revealed that injection of human erythrocytes into the starfish Asterias rubens leads to a decrease of the number of coelomocytes in 24–96 h and to an increase of their specific production of active oxygen forms in 96–120 h after the HE injection. In coelomic fluid of Asterias rubens the presence of peroxidase activity is established. The circulation time of human hemoglobin released from erythrocytes in coelomic fluid of these animals does not exceed 24 h. It is suggested that the cellular defense reactions are the major factor of the starfish congenital immunity.__________Translated from Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, Vol. 41, No. 2, 2005, pp. 107–113.Original Russian Text Copyright © 2005 by Kudryavtsev, D’yachkov, Kazakov, Kanaikin, Kharazova, Polevshchikov.