Morphometric analysis of the larval branchial chamber in the dragonfly Aeshna cyanea Müller (Insecta, Odonata, Anisoptera)

The aquatic larvae of anisopteran dragonflies possess tracheal gills located in the rectum. Using stereological methods, we estimated the morphometric diffusing capacity for oxygen (D(MO2)) across the gill epithelium, i.e., from rectal water to the gill tracheoles, in the larvae of Aeshna cyanea. A 271-mg larva has a total branchial surface area of approximately 12 cm(2). Tracheoles make up 6% of the epithelial volume of the gills; the harmonic mean thickness of the water-tracheolar diffusion barrier is 0.27 microm and consists mainly of cuticle. The calculated D(MO2) is 23.0 microl min(-1) g(-1) kPa(-1), which, using published values for oxygen consumption in a similar species, would result in a mean driving pressure of 0.2 kPa at rest and 1.3 kPa during activity. Since these driving pressures are similar to those reported for other arthropods, we conclude that the D(MO2) of the gill is not rate-limiting for aerobic metabolism in Aeshna cyanea larvae. J Morphol. 261:81-91, 2004.     

Histologie und feinstruktur der larvalen kiemenkammer von Aeshna cyanea Müller (Odonata: Anisoptera)

Zusammenfassung Ein großer Teil des Rectums der Larven von Aeshna cyanea wird von der Kiemenkammer eingenommen. Hier ist das Epithel im Bereich der Kiemenstreifen modifiziert. Ein Kiemenstreifen besteht aus Haupt- und Querfalten. Basale Teile der Querfalten (Basalwülste) werden von Zellen gebildet, die morphologisch einem transportaktiven Epithel ähneln (unter der Cuticula apikaler Mikroleistensaum mit zahlreichen Mitochondrien, basales Labyrinth). Die aus zwei Basalwülsten hervorgehenden Kiemenblättchen, locker aufeinanderliegende, einschichtige Epithelien, zeigen Strukturmerkmale respiratorischer Epithelien (niedrige Zellen, reiche Tracheen- und Tracheolenversorgung in Invaginationen der basalen Plasmamembran, die bis nahe unter die Cuticula reichen). Man darf daher annehmen, daß diesem Enddarmabschnitt neben der Respiration auch ionentransportierende Tätigkeit zukommt.

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Histology and fine structure of the larval branchial chamber in Aeshna cyanea Miller (Odonata: Anisoptera)

Summary In larval Aeshna cyanea the anterior portion of the rectum has been modified into a branchial chamber with six gill folds (corresponding to the pads in the posterior rectal portion). A gill fold is made up of a longitudinal main fold with obliquely placed transverse folds at each side. Each transverse fold consists of two basal pads enclosing fat body cells. The cells of the basal pads show some details reminiscent of transporting epithelia (e.g. apical and basal infoldings, numerous mitochondria). The apical parts of the pads are constricted into lamellae projecting into the rectal lumen. These lamellae are typical respiratory epithelia, which consist of two very flat layers with extracellular tracheoles arranged in invaginations of the basal plasma membrane just beneath the cuticle. Thus it may be assumed that this part of the rectum is concerned with respiration as well as absorption of ions.

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Abkürzungen äE äußere Epicuticula - Bl Basallamelle - Bw Basalwulst - Cu Cuticula - cM circuläre Muskulatur - Dl Darmlumen - En Endocuticula - Fk Fettkörper - Gl Glykogen - hC homogene Cuticula - Hc Hämocoel - Hf Hauptfalte - iE innere Epicuticula (=dense layer) - Kb Kiemenblättchen - Ks Kiemenstreifen - Mi Mitchondrium - Mt Mikrotubuli - Mu Muskeln - Mvb Multivesikularörper - N Kern - Pi Pigmentgranum - Qf Querfalte - Re Rectumepithel - RER rauhes endoplasmatisches Reticulum - sL subcuticulare Lage - TR Trachee - Tr Tracheole Herrn Professor Dr. R. Altevogt, Münster, zum 50. Geburtstag gewidmet.     

Development,cytology, lipid storage and motility of the Malpighian tubules of the nymphal dragonfly,Aeshna cyanea (Müller) (Odonata : Aeshnidae)

The Malpighian tubules of nymphal Aeshna cyanea (Odonata : Aeshnidae) were examined by light and electron microscopy. The 1st-instar nymphs have only 3 branchless tubules. With proceeding nymphal stages, these lengthen and branch. Also, additional tubules bud from the gut and show the same pattern of growth and branching, until in the final instar up to 21 separate tufts of branched tubules are present. A serpentine trachea/tracheole and a cross-striated muscle are helically wound around each tubule in close apposition. Isolated tubules show twisting movements for several days. Contraction of the muscle is responsible for fast coiling movements, while the slow decoiling movements probably depend on elastic deformations of the accompanying trachea, the basal lamina and the tubule cells, the latter showing an elaborate cytoskeleton and multiple adhesive junctions.The tubular epithelium consists of 4 types of cells. The distal segment is composed of ion transporting cells and terminates with a short, solid tip segment of undifferentiated cells. The intermediate segment consists of lipid cells which are densely filled with triglyceride droplets, as revealed by thin layer chromatography. Lipid cells are already present in the 1st instar before the nymphs have taken up any food. In later instars, the renal lipid content varies to some extent with the nutritional state and is nearly depleted during metamorphosis. The proximal segment is the region of tubular branching and may be conceived as the collecting duct of each tuft. Its epithelium consists of mucocytes.     

Übersprungputzen bei der Larve vonAeschna cyanea Müll. (Odonata)

Zusammenfassung 1. Wird bei der Larve vonAeschna cyanea Müll. zunächst das beim Beutefangverhalten auftretende Übersprungputzen durch häufiges Auslösen ermüdet, so nimmt die Anzahl der anschließend auslösbaren Schnappreaktionen ab mit der Zunahme der Anzahl der Übersprungputzhandlungen.2. Für eine Erklärung reicht die Übersprunghypothese, wenn sie in zu einfacher Form oder ausschließlich herangezogen wird, nicht aus.

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Displacement preening in the larva ofAeschna cyanea Müll. (Odonata)

Displacement preening in larvae ofAeschna cyanea is performed in the course of searching behaviour whenever a prey-dummy is suddenly removed. Experiments with a group of animals of the same age, size and origin were carried out to show whether or not there is an influence of displacement preening on the prey-catching behaviour performed afterwards. In control tests all animals showed the snapping-response to a certain dummy 700 to 900 times. After a high number of preening actions, a reduction of the snapping activity was observed. The fact that the displacement preening, in contrast to that of the autochthonous preening, influences the prey-catching activity can be interpreted by a sparking-over of the specific action potential; but one should avoid taking a too simple approach to the conception of a sparking-over by the surplus-hypothesis.

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Herrn Professor Dr.Adolf Bückmann zum 65. Geburtstag in Verehrung gewidmet.

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Mit Hilfe der Deutschen Forschungsgemeinschaft.     

The localization of inorganic elements,particularly vanadium and sulphur,in haemolymph from the ascidians Ascidia mentula (Müller) and Ascidiella aspersa (Müller)

Vanadium and sulphur were found by X-ray microanalysis in three different cell types, morula cells, signet-ring cells, and granular amoebocytes, from each of the ascidians Ascidia mentula (Müller) and Ascidiella aspersa (Müller). In addition to the cells containing vanadium and sulphur a few vacuolar cells containing high concentrations of iron, magnesium and calcium were also found. After fixation in the presence of strontium or barium chlorides another cell type containing large amounts of precipitated sulphur was also found, confirming our previous findings that many haemolymph cells in these species contain large amounts of sulphate only. The presence of these various cell types is discussed in relation to the question of the acidity of ascidian blood cells.     

Analysis of the emergence of the Scarce Chaser Libellula fulva O.F. Müller, 1764 (Odonata: Libellulidae)

The Scarce Chaser (Libellula fulva) is widespread in Europe and in many regions is relatively common. It is also abundant in the lake districts of western and northern Poland. Although the species frequently occurs in large numbers its biology is poorly known. Emergence of L. fulva was investigated in this study as a contribution to the life history of the species. Studies were carried out in the vicinity of Chojno (52°41′ N, 16°12′ E) on the edge of the Notecka Forest in 2003 and 2004. The stem habitat of the population is a stream with the adjacent swamps. Libellula fulva belongs to the spring species sensu Corbet (1999). The emergence of the studied population lasted 19 days (between 18 May and 5 June) and 31 days (between 13 May and 12 June) in 2003 and 2004, respectively. It was characterised by synchronous and, especially in the first phase, mass emergence. The high synchronisation is reflected in the fact that 50% of the population had emerged by the seventh (in 2003) and the fifth (in 2004) days of the process. This is vital to the imagines, which mature and return to the water body as soon as possible to reproduce. Timing of the end of emergence depends on atmospheric conditions during its duration and the conditions preceding the start of emergence on a given day. In the daily course there is a tendency for emergence to finish as soon as possible. Extension of the emergence is synonymous with increased mortality in the dragonfly population caused by predators.     

Contribution of Müller cells toward the regulation of photoreceptor outer segment assembly

The assembly of photoreceptor outer segments into stacked discs is a complicated process, the precise regulation of which remains a mystery. It is known that the integrity of the outer segment is heavily dependent upon surrounding cell types including the retinal pigment epithelium and Müller cells; however the role played by Müller cells within this photoreceptor-specific process has not been fully explored. Using an RPE-deprived but otherwise intact Xenopus laevis eye rudiment preparation, we reveal that Müller cell involvement in outer segment assembly is dependent upon the stimulus provided to the retina. Pigment epithelium-derived factor is able to support proper membrane folding after inhibition of Müller cell metabolism by alpha-aminoadipic acid, while isopropyl beta-D-thiogalactoside, a permissive glycan, requires intact Müller cell function. These results demonstrate that both intrinsic and extrinsic redundant mechanisms exist to support the ability of photoreceptors to properly assemble their outer segments. Our study further suggests that the receptor for pigment epithelium-derived factor resides in photoreceptors themselves while that for permissive glycans is likely localized to Müller cells, which in turn communicate with photoreceptors to promote proper membrane assembly.     

Role of Ca2+ and calmodulin-dependent enzymes in the regulation of glycine transport in Müller glia

Glycine (Gly) is considered an obligatory co-agonist at NMDA receptors. Müller glia from the retina harbor functional NMDA receptors, as well as low and high affinity Gly transporters, the later identified as GLYT1. We here studied the regulation of Gly transport in primary cultures of Müller glia, as this process could contribute to the modulation of NMDA receptor activity at glutamatergic synapses in the retina. We demonstrate that neither glutamate stimulation nor the activation or inhibition of protein kinases A or C modify transport. In order to assess a function for Ca2+ and calmodulin (CaM)-dependent processes in the regulation of Gly transport, we explored the participation of Ca2+ concentration, CaM and Ca2+/CaM-dependent enzymes on Gly transporter activity. ATP and carbachol, known to induce Ca2+ waves in Müller cells, as well as caffeine-induced Ca2+ release from intracellular stores stimulated transport, whereas Ca2+ chelation by BAPTA-AM markedly reduced transport. CaM inhibitors W-7, ophiobolin A, R-24571 and trifluoperazine, induced a specific dose-dependent inhibition of transport. The inhibition of CaMKII by the autocamtide-2-related inhibitory peptide or by KN62 caused a decrease in transport which, in the case of KN62, was due to the abolition of the high affinity component, ascribed to GLYT1. Our results further suggest that Gly transport is under cytoskeletal control, as activation of calpain by major increases in [Ca2+]i induced by ionophores, as well as actin destabilization clearly inhibit uptake. We here demonstrate for the first time the participation of CaM, CaMKII and the actin cytoskeleton in the regulation of Gly transport in glia. Ca2+ waves are induced in Müller cells by distinct neuroactive compounds released by neurons and glia, hence the regulation of [Gly] by this system may be of physiological relevance in the control of retinal excitability.     

Spermatological characters of the pseudophyllidean cestode Bothriocephalus scorpii (Müller, 1776)

Spermiogenesis of Bothriocephalus scorpii (Cestoda, Pseudophyllidea) includes an orthogonal development of two flagella, followed by a flagellar rotation and a proximo-distal fusion with the median cytoplasmic process. The fusion occurs at the level of four attachment zones. The presence of dense material in the apical region of the differentiation zone in the early stage of spermiogenesis appears to be a characteristic feature for the Pseudophyllidea. The mature spermatozoon possesses two axonemes of 9+"1" pattern of the Trepaxonemata, nucleus, cortical microtubules, electron-dense granules and crested body. The anterior part of the gamete exhibits a centriole surrounded by electron-dense tubular structures arranged as incomplete spiral. When the crested body disappears, the electron-dense tubular structures are arranged into a ring encircling the axoneme. The electron-dense tubular structures and their arrangement appear to be a specific feature for the clade "Bothriocephalidea". The organization of the posterior extremity of the gamete with the nucleus is described for the first time in the Pseudophyllidea.     

Studies on the ovotestis of the slug Agriolimax reticulatus (Müller)

Summary The follicle cells, nurse cells and germinal epithelia, which are closely associated with the oocyte of Agriolimax reticulatus (Müller) during its development in the ovotestis, have been studied using light and electron microscopy. The various secretory, digestive and phagocytic activities of these cells have also been investigated using electron cytochemical tests for oxidisable polysaccharide, acid phosphatase and electron-opaque tracer molecules. The oocyte lies initially between the germinal epithelia and a layer of nurse cells but, as oocyte vitellogenesis proceeds, it becomes encapsulated by a layer of follicle cells. Both the follicle and the nurse cells are active in secretion and digestion and contain Golgi apparatus, granular endoplasmic reticulum and acid phosphatase-rich digestive vacuoles. The significance of these activities is discussed in relation to oocyte vitellogenesis, secondary envelope formation and the digestion and recycling of cellular material.     

Possible sites of ultrafiltration in Tubifex tubifex Müller (Annelida,Oligochaeta)

Summary The endothelia of Tubifex tubifex Müller consist of myoendothelial cells, chloragocytes, or podocytes. The latter seem to occur only as windows on the ventral vessel which has an endothelium of myoendothelial cells elsewhere. The podocytes are large cells, with several processes on the inner side which ramify into several pedicels. These are aligned upon the outside of the basement membrane which lines the inside of the endothelium. The gaps between adjacent pedicels are about 40 nm wide. In capillaries fenestrated endothelia occur with irregular spacings measuring up to 0.4–1 m. A diaphragm in podocytes or capillary fenestrations do not seem to exist. The basement membrane is the only continuous layer lining the blood vessels and capillaries of Tubifex with a rather uniform diameter in the range of 50 nm. It is the only permeability barrier between blood and coelomic fluid.     

Studies on the ovotestis of the slug Agriolimax reticulatus (Müller)

Summary The ovarian oocytes of Agriolimax reticulatus (Müller) have been studied by light and electron microscopy and electron cytochemistry. The development of the oocyte in the ovotestis may be divided into three stages.During Stage I the oocyte cytoplasm contains mainly ribosomes and also strands of endoplasmic reticulum, scattered mitochondria and Golgi systems. The nucleus contains both a paranucleolus and an eunucleolus. By Stage II the oocyte has enlarged, especially in a plane parallel to the basement membrane. In addition to the above mentioned organelles, the cytoplasm contains lipid, glycogen and early yolk platelets. During Stage III, the oocyte continues to enlarge, but mainly in a plane perpendicular to the basement membrane. A considerable degree of cytoplasmic differentiation has also taken place. The plasma membrane of the oocyte has become specialized with the appearance of a polysaccharide-rich glycocalyx, microvilli and pinocytotic tubules. Elsewhere, much of the background cytoplasm, containing Golgi-derived, polysaccharide and acid phosphatase-rich multivesiculate bodies, lipid and glycogen, is sequestered by smooth membranes and ultimately fuses with the growing yolk platelets. The nucleus contains an amphinucleolus, characteristic of many gastropods.The findings of this study are discussed in relation to results from other studies on oogenesis.     

Biology of Pseudopotamilla reniformis (Müller 1771) in the White Sea,with description of asexual reproduction

The tube-dwelling polychaete Pseudopotamilla reniformis (Sabellidae) forms dense and complex aggregations of flexible tubes on hard substrates in the subtidal zone of the White Sea. No sexual reproduction was observed in this study and recruitment appeared to be due to asexual reproduction by architomy in winter, from October to March. The posterior part of the abdomen undergoes spontaneous fission into from 2 to 4 fragments and depending on their position, the fragments regenerate their anterior ends or both anterior and posterior ends. Regeneration in P. reniformis takes place via a combination of epimorphosis (replacement of missing parts by cell proliferation and the growth of new tissue) and morphallaxis (the remodelling of pre-existing structures without cell proliferation). The morphogenetic events during regenerative restoration include de novo formation of branchial crown, formation of thoracic segments and restoration of the posterior end. Asexual reproduction appears to play a crucial role for formation of P. reniformis aggregations and is very important for the population in the White Sea, at the margin of the species’ range.     

Comparative studies of the carotenoids in some fauna of the gullmar fiord (Bohuslän,Sweden). IV-Ascidiella aspersa (O. F. Müller), (Tunicata).

The author investigated the presence of various carotenoids in Ascidiella aspersa (O. F. Müller), (Tunicata) by means of columnar and thin-layer chromatography.The investigations revealed the presence of the following carotenoids: cynthiaxanthin (2.9%), capsanthin (32.6%), capsorubin (4.8%), capsorubin-like (15.2%), astaxanthin (4.7%) and astacene (39.8%).     

Fish-induced variation in abdominal spine length of Leucorrhinia dubia (Odonata) larvae?

The lengths of dorsal and lateral spines on abdominal segments 4, 6 and 9 were measured on last-instar larvae of Leucorrhinia dubia (Odonata) from seven natural lentic systems containing fish and nine systems lacking fish. Larvae from systems with fish had significantly longer spines than larvae from systems without fish. In contrast, lake/pond area and pH had no effect on the length of spines. The length of the spines was not correlated with larval size, but there was a high correlation between the length of the three spines measured. Also, abundances of L. dubia larvae differed between systems, being significantly lower in systems with fish. Laboratory experiments showed that perch (Perca fluviatilis) handled long-spined larvae for significantly longer times than short-spined larvae. The results suggest that fish predators may induce morphological defences in insects.     

Role of Müller glia in neuroprotection and regeneration in the retina

Glial cells are thought to protect neurons from various neurological insults. When there is injury to retina, Müller cells, which are the predominant glial element in the retina, undergo significant morphological, cellular and molecular changes. Some of these changes reflect Müller cell involvement in protecting the retina from further damage. Müller cells express growth factors, neurotransmitter transporters and antioxidant agents that could have an important role in preventing excitotoxic damage to retinal neurons. Moreover, Müller cells contact to endothelial cells to facilitate the neovascularization process during hypoxic conditions. Finally, recent studies have pointed to a role of Müller cells in retina regeneration after damage, dedifferentiating to progenitor cells and then giving rise to different neuronal cell types. In this article we will review the role of Müller glia in neuroprotection and regeneration after damage in the retina.     

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.