Electron microscopic studies onNosema mesnili Paillot (Microsporidia: Nosematidae) infecting the Malpighian tubules ofPieris canidia larva

Summary An entire coding region of theCDC24/CLS4 gene and its truncated derivatives were overexpressed in yeast cells under the control of theGAL1 promoter. Western blotting analysis of the yeast cell lysates showed that the CDC24/CLS4 protein (Cdc24p) was induced to reach its maximum level after 9 h incubation of the cells in galactose medium. Overexpression of Cdc24p within the cells caused the morphological change, accumulating large spherical unbudded cells which exhibited actin cytoskeleton disturbed, chitin delocalized on the cell surface, and cell viability decreased. Multiple nuclei were observed in these cells, indicating that only budding cycle but not nuclear division cycle is blocked by the overproduction of Cdc24p. In order to identify the region of Cdc24p responsible for the growth inhibition, several truncatedCDC24 genes were expressed. Surprisingly, overexpression of fragments either containing the C-terminal 76 amino acid residues or deleting the same region inhibited cellular growth. This suggests that Cdc24p contains multiple functional domains for its tasks, likely cooperating signals of bud positioning and bud timing.     

Electron microscopic studies on a unique cytokinetic structure in tetrasporocytes of the red algaHarveyella sp. (Cryptonemiales,Choreocolaceae)

Summary A unique type of cytokinesis is described in tetrasporocytes of the alloparasitic red alga,Harveyella sp. Cytokinesis takes place immediately after four post-meiotic nuclei are formed and may result from coalescence of endoplasmic reticulum cisternae. As a result of this cisternal fusion, a double-membraned cleavage channel is formed. This channel lacks wall material and is usually oriented tetrahedrally. Membranes of the peripheral portions of the cleavage channel fuse with the existing tetrasporocyte plasmalemma, delimiting four tetraspores. Subsequently, wall material is secreted within the preformed cleavage channels to form a continuous tetraspore wall between adjacent tetraspores. Wall secretion usually occurs in a centrifugal direction, beginning at the juncture of the cleavage channels, but it also may be random or centripetal. Dictyosome activity is absent during the first wall secretion stage but contributes to secondary wall deposition.Portions of this study were conducted at the University of Washington's Friday Harbor Laboratories, Friday Harbor, Washington.     

Neurochemical characterization of nervous elements innervating the body wall of earthworms (Lumbricus, Eisenia): immunohistochemical and pharmacological studies

The distribution and chemical neuroanatomy of nervous elements and certain pharmacological–physiological characteristics of the innervation of the body wall in earthworms are described. Solitary sensory bipolar cells can be found among the epithelial cells. These bipolar cells contain serotonin, tyrosine hydroxylase, histamine, gamma-amino-butyric acid (GABA), Eisenia tetradecapeptide, proctolin or rhodopsin in various combinations. In the body wall, the plexus submuscularis is composed of nerve fibres only, whereas the plexus subepithelialis and muscularis also contain solitary nerve cells. These cells display histamine, GABA or neuropeptide Y immunoreactivity. The fibres of the three plexuses are reactive to serotonin, histamine, Eisenia tetradecapeptide, proctolin, GABA and neuropeptide Y antibodies. FMRFamide-immunoreactive fibres of the plexus muscularis originate from the central nervous system, whereas axons containing the other studied molecules are derived from both peripheral and central structures. High pressure liquid chromatography assays have revealed serotonin, dopamine and histamine in the body wall. Contractions of the body wall musculature can be elicited with serotonin and FMRFamide. Serotonin-evoked contractions are suppressed by the application of GABA. Serotonin acts both directly on the muscle cell receptors and indirectly through initiating transmitter release from the nervous elements, whereas the FMRFamide-induced contractions seem to be mediated through the muscle cell receptors only. The pharmacological profiles of the serotonin and GABA receptors resemble those of the vertebrate 5-HT₃ and GABA_B receptor types. Our findings indicate that both the sensory and efferent system of the annelid body wall operate by means of a variety of neuroactive compounds, suggesting a complex role of signalling systems in the regulation of this organ.This work was supported by the Hungarian Scientific Research Fund (OTKA; grant nos. T 34106 and T 34160). Márta Wilhelm is in receipt of a János Bolyai Scholarship.     

Electron microscopic study on the epiplexus (Kolmer) cells of the cat choroid plexus

Summary A light and electron microscopic study was made of the epiplexus (Kolmer) cells of the cat choroid plexus. These polymorphic, motile cells were typically found juxtaposed to the ventricular surface of the choroidal epithelium. They have many ultrastructural features in common with free macrophages of other systems, namely, an indented nucleus with condensed chromatin, sparse mitochondria and endoplasmic reticulum, free ribosomes, multiple Golgi elements, microtubules, coated surface invaginations and microvesicles, and numerous membrane-limited vacuoles and lysosomal dense bodies. A unique feature of epiplexus cells is the manner in which they are anchored to the choroidal epithelium by the invagination of their surfaces by epithelial cell microvilli and cilia.Electron dense tracer particles (biological India ink, Thorotrast, ferritin) injected into the cerebral ventricles were ingested rapidly by epiplexus cells. Uptake of the particles was by way of coated surface invaginations which produced coated cytoplasmic microvesicles. Particle-containing microvesicles subsequently fused with each other and presumably also with pre-existent cytoplasmic vacuoles and lysosomal dense bodies to form storage vacuoles (phagosomes phagolysosomes and residual bodies).Present evidence suggests that epiplexus cells are of hematogenous origin. Under certain conditions these cells may detach from the surface of the choroid plexus to become free-floating cells in the various cerebrospinal fluid compartments of the brain.This investigation was supported by USPHS research grants 1-K04 HD20871, 5 R01 HD 02616 and NB-04456.     

Ultrastructure of aesthetasc innervation and external morphology of the lateral antennule setae of the spiny lobster Panulirus interruptus (Randall)

Summary Six types of setae and one type of cuticular depression were examined on the lateral antennule of the spiny lobster Panulirus interruptus using scanning electron microscopy. The organization and ultrastructure of the innervation of the most numerous setal type, the aesthetasc, were investigated using light-and transmission electron microscopy.Each aesthetasc is innervated by approximately 300 bipolar neurons whose sensory dendrites penetrate the hair and extend toward the tip, and whose axons project towards the central nervous system. The neuronal somata and two types of glia form a cluster within the antennular lumen. The inner sheath-cell somata encircle the dendritic tract distal to the sensory somata. These cells appear to extend distal processes which wrap the dendritic tract to the base of the aesthetasc. Elongate outer sheath cells are interposed between the glia-wrapped dendritic tract and the hypodermis which underlies the antennule cuticle. A continuous investment of neural lamella separates the hypodermis, the entire cluster of somata, and sensillar nerve from the antennule lumen. The organization of the neuronal somata and their association with outer and inner sheath cells in this marine species appear similar to those of crustaceans from freshwater and terrestrial habitats.     

Ultrastructure of the neurosecretory system of the Colorado potato beetle,Leptinotarsa decemlineata (Say)

Summary The ultrastructure of seven types of neurosecretory cells (NSC) in the medial and lateral groups of the protocerebrum is described. The differences among cell types established earlier by light microscopy parallel differences in size and appearance of the neurosecretory particles observed in electron micrographs. No relationship was found between the affinity for Gomori's paraldehyde fuchsin stain and the nature of the particles.The secretions of the A-, A₁-, and C-types of NSC of the medial group are characterized by electron-dense neurosecretory granules of 1250 Å dia., medium-dense granules of 2100 Å, and electron-lucent vesicles of 1700 Å, respectively. The L-type NSCof the lateral group contain smaller (1300 Å) or larger (1700 Å) neurosecretory granules. The medial B- and E-types of NSC and the lateral L_B-type contain granulated vesicles (1200 Å) of the same appearance. These cell types differ in other respects and most likely have separate functions.The author wishes to thank the Laboratory of Virology of the Agricultural University for the use of the electron microscope, Mr. J. Groenewegen and Miss J. van Rinsum for technical assistance, and Professor J. Lattin for correcting the English text. Part of the work has been done while the author was in the service of the Netherlands Organization for the Advancement of Pure Research (ZWO, grant 942-48), and the National Council for Agricultural Research (TNO).     

Ultrastructure of the taste disc in the red-bellied toad Bombina orientalis (Discoglossidae,Salientia)

The taste disc of the red-bellied toad Bombina orientalis (Discoglossidae) has been investigated by light and electron microscopy and compared with that of Rana pipiens (Ranidae). Unlike the frog, B. orientalis possesses a disc-shaped tongue that cannot be ejected for capture of prey. The taste discs are located on the top of fungiform papillae. They are smaller than those in Ranidae, and are not surrounded by a ring of ciliated cells. Ultrastructurally, five types of cells can be identified (mucus cells, wing cells, sensory cells, and both Merkel cell-like basal cells and undifferentiated basal cells). Mucus cells are the main secretory cells of the taste disc and occupy most of the surface area. Their basal processes do not synapse on nerve fibers. Wing cells have sheet-like apical processes and envelop the mucus cells. They contain lysosomes and multivesicular bodies. Two types of sensory cells reach the surface of the taste disc; apically, they are distinguished by either a brush-like arrangement of microvilli or a rod-like protrusion. They are invaginated into lateral folds of mucus cells and wing cells. In contrast to the situation in R. pipiens, sensory cells of B. orientalis do not contain dark secretory granules in the perinuclear region. Synaptic connections occur between sensory cells (presynaptic sites) and nerve fibers. Merkel cell-like basal cells do not synapse onto sensory cells, but synapse-like connections exist between Merkel cell-like basal cells (presynaptic site) and nerve fibers.     

The distribution of and interactions between GABA-immunoreactive and non-immunoreactive processes presynaptic to afferents from campaniform sensilla on the trochanter of the locust leg

Summary Campaniform sensilla on the trochanter of the mesothoracic legs of the locust were backfilled with cobalt salts or horseradish peroxidase for light and electron microscopy. The distribution of the terminal branches of afferent neurones in the thoracic ganglia were described from wholemount preparations and from thick slices through the ganglia. Ultrathin sections of identified branches were processed with GABA antibodies using a post-embedding immunogold technique and examined in the electron microscope. Input synapses were observed on fine varicose branches in all regions of the terminal arborisations close to the sites of afferent output. The major branches neither make nor receive synapses. Seventy-two percent of the input synapses are made by processes immunoreactive for GABA. Immunoreactive and non-immunoreactive processes synapse onto afferent terminals in close proximity. In some instances GABA-immunoreactive processes presynaptic to an afferent are also presynaptic to a non-immunoreactive presynaptic processes strongly suggesting that different presynaptic influences can interact directly with each other.     

A microscopic investigation of enamel in wombat (Vombatus ursinus)

Summary A study of the enamel of continuously growing Vombatus ursinus molars was carried out using the techniques of light microscopy, hardness testing, scanning electron microscopy and transmission electron microscopy. From the erupted end to within 8 mm of the growing end, mature enamel was observed and it was found that between comparable areas there were no significant ultrastructural differences in enamel; however, small (12nm diameter), loosely packed needle-like crystals characteristic of developing enamel were observed near the growing end. Mature enamel was found to consist of three optically-translucent regions interleaved with two opaque regions. Opaque enamel was softer than translucent enamel. The opacity and relative softness characteristic of two of the enamel regions was not related to prism pattern or orientation; it was, however, related to the presence of voids (28 nm diameter) in these regions.     

Electron microscopic study of the mandibular glands of Kalotermes flavicollis Fabr. (Isoptera; Calotermitidae)

Summary The mandibular glands of Kalotermes were examined in different castes. They showed sexual dimorphism in the soldiers and primary reproductives. Moreover, in female soldiers and queens, mandibular gland cells contained numerous crystalline structures of mitochondrial origin. The role of these glands (secretion of saliva or pheromone) is discussed.

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Résumé Les glandes mandibulaires de Calotermes sont étudiées dans les différentes castes. Elles présentent un dimorphisme sexuel chez les soldats et les sexués. Après la mue imaginale, les cellules glandulaires ont toujours une activité sécrétrice. De plus, chez les soldats femelles et les reines, elles contiennent de nombreuses formations cristallines d'origine mitochondriale. Le rôle de ces glandes (sécrétion de salive ou de phéromone) est discuté.

     

The fine structure of distal receptors on the labium of the aphid,Brevicoryne brassicae L. (Homoptera)

Summary Short peg receptors located at the distal tip of the aphid labium have the structure of mechanoreceptors. Each peg is innervated by a single sensory nerve which is anchored eccentrically to a basal cuticular tube and terminates in electron-dense material in the base of the peg. The arrangement and eccentric insertion of the eight pegs in the labial wall on one side of the stylet groove, with the eccentric insertions of their innervating neurones, provide a mirror image of the receptors on the opposite side. On the basis of a comparison of the structure of these receptors with that of tactile receptors for which electrophysiological data on sensitivity are available, it is possible to predict that the receptors detect both surface contact (pressure) and surface profile; and that the bilateral symmetry in the receptor arrangement facilitates the detection of vein contours which are preferred settling sites on the leaf. The structure of the dendritic terminal and its insertion is that of a well reinforced cytoskeleton designed to transmit tension to the cell membrane, in agreement with the concept that transduction is a membrane related phenomenon. The distal microtubules, fifty per-cent of which originate as well as terminate in the tubular body, are packed in electron-dense material which binds to the cell membrane. The membrane in turn is attached to cuticular components of the receptor. Abrupt changes in dimension of the dendritic outer segment may be designed to modulate the conduction of a membrane potential. On the other hand, lack of continuity in the microtubules makes these organelles poor candidates for the transduction of excitation from a distal site of stimulation to a proximal region.Supported by operating grants Nos. A 6063 and A 9856 from NRCC     

Scanning electron microscopy of the muscle system of Hydra magnipapillata

The fine structure of the ectodermal and endodermal muscle layers of Hydra magnipapillata has been analyzed by scanning electron microscopy after hydrolytic removal of the mesoglea with NaOH and subsequent exposure of the basal and lateral aspects of the layers by mechanical dissection. The ectodermal muscle layer consists of fibrous processes of epithelial cells extending longitudinally to the body axis, whereas the endodermal muscle layer comprises cells with hexagonal bases and several strands of myonemes oriented circularly. In each layer, the muscular elements tightly interdigitate, extending a continuous muscle sheet along the mesoglea. The ectodermal and endodermal muscle sheets communicate with each other via foliate microprojections penetrating the mesoglea. On the lateral aspect of the ectodermal epithelium, spiny nerve fibers run along the upper surface of the muscle processes. The spines are often attached to muscle processes, suggesting that the former monitor muscle contraction. Nerve fibers occasionally come into contact with the mesoglea through narrow gaps between the muscle processes. In the hypostomal ectoderm, a small spindle-shaped cell, probably sensory in nature, extends an apical cilium and a long basal process.     

Electron microscopic and histochemical investigations of the atretic oocyte of Perca fluviatilis L. (Teleostei)

Summary The regression of atretic oocytes in Perca fluviatilis was studied by histochemical, light and electron microscopic methods. The course of regression can be divided into three stages, the first two comprising the dissolution of the atretic oocyte and its phagocytosis by the granulosa cells of the follicular epithelium, and the third stage consisting of the dissolution of the granulosa cells themselves. The ultrastructure in all three stages shows only features related to phagocytosis and lysosome formation. In particular, there is no agranular endoplasmic reticulum formed within the phagocytically active granulosa cells, nor is there any 3-hydroxysteroid dehydrogenase activity (3-HSD). Large yellow-orange pigments, formed during the third stage of regression, are ascribed to a relative deficiency of lysosomes in lipid digestion, and do not result from a preceeding steroid-synthesising phase as in mammalian corpora lutea. Thus, the atretic oocyte of P. fluviatilis is considered not to give rise to a corpus luteum formation with endocrine function, but merely represents a degenerative structure.     

Electron microscopic study on the larval and adult corpus allatum of Oncopeltus fasciatus dallas (insecta,heteroptera)

Summary 1. The ultrastructure of the corpora allata of last larval instars and adults of Oncopeltus was studied. The unpaired gland undergoes submicroscopic alterations and shows signs of degradation in old animals. The organ is partly covered and penetrated by corpus cardiacum tissue. Axons with different types of neurosecretory granules form synaptoid contacts with the corpus allatum cells.2. Dark and light gland cells can be differentiated on account of the degree of electron density. The former predominate during the last larval stage and in the young imago, the latter in mature males and females. It is highly probable that the light cells are the active (i.e. hormone producing) ones and the dark cells the inactive ones.3. The active cells are characterized by rough endoplasmatic reticulum (often in whorls), small amounts of smooth endoplasmatic reticulum and many multivesicular bodies. Abundant free ribosomes, a not particularly well developed Golgi apparatus, dense bodies, and cytolysomes are present in active and inactive cells.4. The nuclei contain one to four prominent and variously shaped nucleoli, which show differences between adult males and females with respect to their location in the nucleus.5. The corpus allatum cells of Oncopeltus are obviously engaged in extensive protein synthesis. Tangible structural indications for the manufacture of juvenile hormone were not observed. Possible sites of hormone release are discussed.This study was made possible by a fellowship and grants from the Deutsche Forschungsgemeinschaft and was supported by research grants, administered by Prof. Scharrer, from the U.S.P.H. Service (NB-05219; NB-00840 and NS-07512). Present address of author: Institut für Allgemeine Zoologie der Johannes Gutenberg-Universität, D-6500 Mainz, Saarstraße 21, Bundesrepublik Deutschland.I am indebted to Prof. Scharrer for guidance and criticism. I also wish to express my appreciation to Mrs. S. Wurzelmann and Mr. S. Brown for their excellent technical assistance.     

六斑月瓢虫触角及感觉器扫描电镜观察

[目的] 明确六斑月瓢虫雌雄成虫触角感觉器种类、分布及形态特征。[方法] 利用扫描电子显微镜对六斑月瓢虫雌、雄成虫触角形态及触角感受器超微结构进行观察。[结果] 六斑月瓢虫成虫触角由柄节、梗节和鞭节组成,柄节长度与宽度显著大于梗节长度与宽度;鞭节分为9个亚节,末端3节横向膨大呈锤状。雌雄成虫触角上共有8种感觉器:刺形感觉器(SC)、毛形感觉器(ST)、锥形感觉器(SB)、腔形感觉器(CaS)、钟形感觉器(CS)、哑铃形感觉器(DS)、香肠形感觉器(SS)及B?hm氏鬃毛感觉器(BB)。以毛形感觉器和刺形感觉器分布最广,遍布触角;B?hm氏鬃毛仅存在于触角柄节与梗节;触角鞭节第9亚节顶端密布7种触角感觉器。六斑月瓢虫雌雄成虫触角长度、触角感觉器类型及分布无显著差异。[结论] 六斑月瓢虫成虫触角上共有8种感觉器,其触角可能具有感知机械刺激、识别化学信息素及感受温湿度变化的作用。本研究为进一步了解六斑月瓢虫触角与其行为间的关系提供基础资料。     

Ultrastructural localization of acetylcholinesterase in the synganglion of the tick,Dermacentor variabilis (Say)

Summary Light- and electron-microscopic enzyme cytochemistry was used to localize acetylcholinesterase (AChE) activity in the synganglion (brain) of the tick Dermacentor variabilis. High AChE activity was observed throughout the neuropil as well as adjacent to most neuronal perikarya. Intracellular activity was not observed by light microscopy. By electron microscopy, reaction product was localized at the plasma membrane of glia and neurons. Enzyme activity was not associated with the olfactory globuli neurons. In other types of neurons, small amounts of reaction product were observed in the Golgi apparatus and nuclear envelope. Large neurosecretory neurons contained activity that appeared to be associated with deep invaginations of the plasma membrane as well as intracellular membranes. AChE activity was also associated with processes of both neurons and glia. In most peripheral nerves AChE activity was associated with virtually all axons. Clearly then, AChE is associated with glia and non-cholinergic neurons as well as with presumed cholinergic neurons. The widespread localization and large amounts of AChE in the tick brain exceeds that reported for other invertebrates and vertebrates. As has been suggested for other animals, AChE in the tick brain may have functions in addition to its known role in cholinergic neurotransmission.     

Electron microscopic study of Burkholderia cepacia biofilms

Using the methods of transmission electron microscopy, the structure of the biofilms formed by the bacterium Burkholderia cepacia (clinical isolate and mutants with an increased and decreased ability to produce biofilm) were investigated. The biofilms were obtained on a liquid nutrient medium or on an abiotic surface (polystyrene). It has been demonstrated that the cultures of the studied strains differ in some morphological and functional characteristics. In biofilms, changes in the size and submicroscopic organization of all the components of bacterial cells occur. Staining biofilms with ruthenium red revealed the presence of exopolysaccharides in the intercellular space. The differences in the ultrastructure of bacterial films formed on nutrient medium and abiotic surfaces were demonstrated.     

Histochemical and ultrastructural studies on the enterochromaffin-like cell in the gastric mucosa of the opossum Didelphis albiventris (Marsupialia)

Summary An ultrastructural study of enterochromaffin-like (ECL) cells in the gastric mucosa of the white-belly opossum Didelphis albiventris (Marsupialia) was carried out. In parallel, histochemical methods were used at the light-microscopical level to demonstrate argentaffin cells, argyrophilic cells, and serotonin- and histamine-immunoreactive elements. Argentaffin and serotonin-immunoreactive cells were scattered, and argyrophilic cells were numerous, within the full thickness of the mucosa. Argyrophilic cell distribution was similar to that of histamine-immunoreactive elements. At the electron-microscopical level, the oxyntic mucosa of D. albiventris presented endocrine cells with secretory granules morphologically similar to those of the ECL cell of eutherian mammals. However, in this marsupial, the ECL cell exhibited a variable mixture of two distinct types of secretory granules: (1) granules with the morphological appearance of the eutherian ECL cell, and (2) granules morphologically similar to those of the eutherian enterochromaffin (EC) cells. Based on this morphological pattern of the ECL cell granules, it is proposed that in the oxyntic mucosa of the opossum D. albiventris, the EC and ECL cells represent distinct steps in the same line of cell differentiation; the ECL cell should also be a site of histamine storage.