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 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.     

Comparison of morphological characters in Irish and English populations of the acanthocephalan Pomphorhynchus laevis (Müller, 1776)

Pomphorhynchus laevis is believed on ecological evidence to exist as three strains in the British Isles. However, the strains have never been shown to be capable of being distinguished using morphological characters. A morphological comparison was made between a sample of P. laevis from Salmo trutta in L. Feeagh in the west of Ireland and a sample from Leuciscus cephalus in R. Culm in the south of England. The length and width of the trunk, neck, bulb, proboscis and hooks were measured. The number of hooks per row, the number of rows and the positions of the stoutest and longest hooks were also recorded. A Principal Components Analysis based on the morphological measurements confirmed the separation of the two populations and showed that two characters successfully identified the populations: the position of the stoutest hook and the ratio of numbers of anterior to posterior hooks.     

Short-term feeding of Temora longicornis Müller in the laboratory and the field

Short-term feeding (min to h) of the copepod Temora longicomis (Müller) was investigated in the laboratory and in the field by monitoring the increase in gut fluorescence of adult females. Laboratory feeding studies using the diatoms Thalassiosira pseudonana (Hust.), T. weissflogii (Hust.) and Ditylumbrightwellii (West) showed that ingestion rates increased with increased food concentration and cell size. The maximum ingestion rate was 9.5 ng chl. equiv. female ⁻¹. h ⁻¹ on a diet D. brightwellii. Clearance rates decreased with increased food concentration and decreased cell size. Maximum clearance rate was 1.9 ml. female⁻¹·h⁻¹ also on a diet of D. brightwellii. On two of three occasions, ingestion and clearance rates in the field were twice as high at night as during the day. Night rates were similar to laboratory rates in the chlorophyll range found in the field. Diel differences in feeding activity in the field appeared to have been behaviorally induced rather than being related to vertical migration of individuals into food rich layers.     

Micro-structure and chemical composition of vateritic deformities occurring in the bivalve Corbicula fluminea (Müller, 1774)

Vateritic deformities occurring in the invasive heterodont bivalve Corbicula fluminea from several locations in the UK were characterised in detail for the first time using scanning electron microscopy, X-ray diffraction and different geochemical techniques (electron microprobe, ICP-AES, and mass spectrometry). Large volumes of vaterite are produced abnormally in the animals' shells in the form of yellow-green bulges. These are distinguished from the aragonitic parts of the shell by their characteristic micro-structures, content of organic material, trace elemental composition and carbon stable isotope signatures. The most commonly observed micro-structures include columnar vaterite, lamellar vaterite and different irregular structures occurring in all parts of the shell. There are indications that organic material is present largely as intracrystalline impurities or nano-scale phases and not as envelopes around microstructural units. These micro-structures are novel, nothing equivalent having yet been described for other vateritic systems. Euhedral vaterite crystals also occur occasionally. The vaterite has generally higher Mg/Ca and lower Na/Ca, K/Ca than the aragonite. In addition, δ13C is also always lower. Microstructural characteristics would suggest loss of biological control probably due to physiological stress(es) inducing the switch to vaterite production. The vaterite might be stabilised by its higher content of organic material and magnesium.     

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.     

Phenotypic variability of a perch parasite-cestode proteocephalus percae (Müller, 1780) (Proteocephalidea) in different parts of the species range

Discrete variability of four P. percae characters of the main cestode functional complexes was identified. Phenotypic diversity of P. percae from different parts of the distribution range was analysed. Research revealed low geographic variability and high stability of the dominant variations in combination with morphometric plasticity. The conclusion was made that the patterns of P. percae morphological variability were shaped by the common fate and long-standing co-evolutionary relations between the parasite and the host--the perch Perca fluviatilis.     

Ionic regulation of the haemolymph in the larvae of the dragonfly Aeshna cyanea (Müller) (Odonata, Anisoptera).

The ionic regulation of the haemolymph of larvae of Aeshna cyanea (Müller) was studied by means of two types of experiments. In the first the change in internal ionic composition was followed as a function of the time spent in a given experimental medium. These experiments led to the conclusion that: 1. the haemolymph composition does not change when larvae are starved in tap water for 10 days; 2. the haemolymph ionic concentrations (Na and Cl) have initial marked increase when the animals are kept in hypertonic media of diluted sea water; after 80 hours however, both concentrations stay constant. In a second series of experiments the internal ionic concentration was compared to a series of different concentrations of external media. From this, the relation between the internal and external ionic concentration was elaborated : in hypotonic media the internal Na and Cl concentrations stay constant, in hypertonic media there is a parallelism between the increase of the external concentration and haemolymph concentration, the internal Na concentration being always slightly hypertonic, the Cl concentration markedly hypotonic. Finally, when larvae are placed in glass distilled water, the internal Na and Cl concentrations begin to decrease after 60 hours.     

Morphology and distribution of Müller cells in the retina of the toad Bufo marinus

We have previously shown that an antibody against neuron-specific enolase (NSE) selectively labels Müller cells (MCs) in the anuran retina (Wilhelm et al. 1992). In the present study the light- and electron-microscopic morphology of MCs and their distribution were described in the retina of the toad, Bufo marinus, using the above antibody. The somata of MCs were located in the proximal part of the inner nuclear layer and were interconnected with each other by their processes. The MCs were uniformly distributed across the retina with an average density of 1500 cells/mm². Processes of MCs encircled the somata of photoreceptor cells isolating them from each other by glial sheath, except for those of the double cones. Some of the photoreceptor pedicles remained free of glial sheath. Electron-microscopic observations confirmed that MC processes provide an extensive scaffolding across the neural retina. At the outer border of the ganglion cell layer these processes formed a non-continuous sheath. The MC processes traversed through the ganglion cell layer and spread beneath it between the neuronal somata and the underlying optic axons. These processes formed a continuous inner limiting membrane separating the optic fibre layer from the vitreous tissue. Neither astrocytic nor oligodendrocytic elements were found in the optic fibre layer. The significance of the uniform MC distribution and the functional implications of the observed pattern of MC scaffolding are discussed.     

Birth and growth of the shortnose guitarfish Zapteryx brevirostris (Müller & Henle, 1841) (Chondrichthyes,Rhinobatidae) in captivity

Endemic to the south-west Atlantic Ocean, the shortnose guitarfish (Zapteryx brevirostris) is a small species, classified as endangered by the IUCN. Although reproduction in captivity has been successful for some species, a range of factors can limit the success of captive breeding programmes for elasmobranchs. In Brazil, the Ubatuba Aquarium was the first public aquarium to reproduce small-sized elasmobranchs. Since 2018, at least five parturition events have been recorded for Z. brevirostris at the institution. From a total of 13 live neonates that rearing was attempted, the mean ± standard error of weight, total length (TL) and disc width at birth were 17.47 ± 1.6 g, 13.25 ± 0.7 cm and 6.53 ± 0.2 cm, respectively. The mean weight as well as mean TL were higher for females at all births, with 26.15 g and 15.07 cm for females in comparison with 17.09 g and 13.94 cm for males. Considering the increasing risk of extinction that the species is facing, age and growth studies are fundamental for the success of conservation, improving the knowledge of Z. brevirostris life cycle so that a more efficient and sustainable management can be carried out. This study provides important data, as well as directions for captive breeding of the species.     

An alternative isomerohydrolase in the retinal Müller cells of a cone-dominant species

Cone photoreceptors have faster light responses than rods and a higher demand for 11-cis retinal (11cRAL), the chromophore of visual pigments. RPE65 is the isomerohydrolase in the retinal pigment epithelium (RPE) that converts all-trans retinyl ester to 11-cis retinol, a key step in the visual cycle for regenerating 11cRAL. Accumulating evidence suggests that cone-dominant species express an alternative isomerase, likely in retinal Müller cells, to meet the high demand for the chromophore by cones. In the present study, we describe the identification and characterization of a novel isomerohydrolase, RPE65c, from the cone-dominant zebrafish retina. RPE65c shares 78% amino acid sequence identity with RPE-specific zebrafish RPE65a (orthologue of human RPE65) and retains all of the known key residues for the enzymatic activity of RPE65. Similar to the other RPE-specific RPE65, RPE65c was present in both the membrane and cytosolic fractions, used all-trans retinyl ester as its substrate and required iron for its enzymatic activity. However, immunohistochemistry detected RPE65c in the inner retina, including Müller cells, but not in the RPE. Furthermore, double-immunostaining of dissociated retinal cells using antibodies for RPE65c and glutamine synthetase (a Müller cell marker), showed that RPE65c co-localized with the Müller cell marker. These results suggest that RPE65c is the alternative isomerohydrolase in the intra-retinal visual cycle, providing 11cRAL to cone photoreceptors in cone-dominant species. Identification of an alternative visual cycle will contribute to the understanding of the functional differences of rod and cone photoreceptors.     

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.     

Variables affecting body burdens of lead,zinc and cadmium in a roadside population of the snailCepaea hortensis Müller

Summary Application of multiple regression analysis to the lead, zinc and cadmium contents of 985Cepaea hortensis collected from a suburban roadside site over two years showed that body weight, age and daylength were the major factors affecting soft tissue body burdens. Other highly significant influences were physiological changes at the juvenile-adult transition, vapour pressure deficit for the two days before sampling, and mean rainfall for the ten days before. Together these factors accounted for 69%, 79% and 87% of the total variance in lead, zinc and cadmium body burdens respectively. Unexplained variance was provisionally ascribed to individual differences in recent diet and digestive activity. The digestive gland contined the majority of the total body burden of each metal and played an important role in lead and zinc elimination. Whereas levels of lead inC. hortensis fluctuated rapidly, zinc was exchanged more slowly, perhaps by active regulation, and cadmium was effectively immobile, accumulating progressively with age. The implications of these results to the predators of land snails, and to terrestrial monitoring programmes, are discussed.     

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.     

Variation in ostracod (Crustacea,Ostracoda) communities in the alluvial valley of the upper Paraná River (Brazil) in relation to substrate

Large river floodplains are convenient model systems to test for variation in animal and plant community structure, as they have a variety of habitats and substrates and are generally dynamic systems through the occurrence of flood pulses with varying intensity. South American floodplain systems furthermore have unique types of substrates, in the form of root systems of floating macrophytes. Here, we investigate the variation in ostracod (small, bivalved crustaceans) communities in relation to substrates and related environmental variables. Sampling was effected in 2004 in the alluvial valley of the upper Paraná River, Brazil, in the wet and dry seasons. Five different substrates, including littoral sediment and four macrophyte species root and leaf systems, in four hydrological systems and a variety of habitat types, were sampled. Fifty-four species of Ostracoda were found. Variation partitioning analysis (RDA) showed that ostracod communities significantly differed between different substrates, mainly between the littoral and plants with small root systems (Eichhornia azurea) on the one hand, and plants with large and complex root systems on the other hand (Eichhornia crassipes and Pistia stratiotes). RDA analyses indicated that the pleuston (biotic communities associated with root systems of floating plants) of E. crassipes comprised more non-swimming species than the pleuston of the smaller roots of P. stratiotes, but species-level Kruskal–Wallis analyses could not detect significant differences between both macrophyte species. Also habitat type and hydrological systems contributed to variation amongst ostracod communities, but less so than the factor substrate. Abiotic factors also contributed to variation, but the ranges of all measured water chemistry variables were narrow. This uniformity in abiotic factors, which might be owing to the occurrence of large flooding events, unites all water bodies, even those that are generally separated.     

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.     

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.     

Distribution pattern of the dinoflagellateCeratium hirundinella (O. F. Müller) Bergh in a reservoir

Horizontal distribution of the dinoflagellateCeratium hirundinella (O. F. Müller) Bergh in the Ishitegawa Reservoir, Ehime Prefecture, Japan, was investigated. Water quality was also surveyed. It was observed that the population ofC. hirundinella exponentially decreased in number from the head of the reservoir to the dam site. Further investigation proved thatC. hirundinella initiated growth at the head of the reservoir, and later gradually expanded downstream. It was found during the period of increase in water temperature that the cell density ofC. hirundinella at the uppermost station exponentially increased.     

Biological studies and phenology of the slug Deroceras reticulatum (Müller, 1774) (Pulmonata: Stylommatophora)

Summary

In the southeast of Buenos Aires Province (Argentina), slugs cause reductions in crop yield by killing seeds and seedlings, by destroying stems and growing points, and by reducing leaf area. Deroceras reticulatum (Müller, 1774) was introduced into Argentina a long time ago, but the biology and ecology of this species in this country are still unknown. The aim of this study was to determine how temperature affects growth rate, reproduction and survival of D. reticulatum and also to assess the seasonal phenology of this species. The life cycle of D. reticulatum was investigated at 12°C, 20°C and at a temperature which alternated between 12°C for 16 h and 20°C for 8 h. The light/dark regime during the experiment was LD: 8/16 h. Slug populations were sampled from grassland located at the Experimental Station of INTA Balcarce. Eggs hatched after 16.3 to 39.8 days and fertility varied between 83.9% and 91.1%. The mean time from hatching to oviposition was longer and fecundity lower at 20°C than 12°C and 12/20°C (p <0.05). The net reproductive rate was 1.49 to 70.53 and the mean generation time varied between the 27 and 30 weeks. In natural conditions, eggs of D. reticulatum were found from early winter until the end of the spring, when soil temperature was 8–15°C and humidity was above 15%. These results indicate that D. reticulatum has one generation per year, univoltine phenology, with peak densities of slugs from the beginning of winter to the end of spring, which coincides with the sowing and emergence of winter and summer crops.     

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.