Membrane specializations in the peripheral retina of the housefly Musca domestica L.

Membrane specializations of the peripheral retina of the housefly (Musca domestica) are revealed in thin sections and freeze fracture/etch replicas. Septate junctions are abundant in corner areas of the pseudocone enclosure bonding: between homologous corneal pigment cells (CPC); between homologous large pigment cells (LPC); between CPC-LPC; between Semper cells (SC); between SC-CPC. Spot desmosomes are present between Semper cells. It is likely that septate junctions function as strengthening adhesions in this area. A new membrane specialization similar to a continuous junction was observed between retinular cells of the same or adjacent ommatidium. This junction has indistinct septa in the 115 A intermembrane cleft and is intermittent in character. When this junction is absent, the apposed cells gape apart. In freeze fracture studies, this junction is characterized by bridges composed of fused membrane particles and randomly arranged particles on the P face, and noncorresponding grooves on the E face. The ridges are elongate and roughly parallel and sometimes they form enclosures. Mitochondria line up along these junctions, often within 90 A of the unit membrane. This membrane specialization has characteristics of tight and continuous junctions. In line with previous findings, we suggest that this junction assists in retinular cell orientation, possibly in enforcing the ommatidial twist and in maintaining localized ionic concentration gradients between retinular cells.     

Interneuronal and glial-neuronal gap junctions in the lamina ganglionaris of the compound eye of the housefly,Musca domestics

Summary The cell-body layer of the lamina ganglionaris of the housefly, Musca domestica, contains the perikarya of five types of monopolar interneuron (L1–L5) along with their enveloping neuroglia (Strausfeld 1971). We confirm previous reports (Trujillo-Cenóz 1965; Boschek 1971) that monopolar cell bodies in the lamina form three structural classes: Class I, Class II, and midget monopolar cells. Class-I cells (L1 and L2) have large (8–15 m) often crescentshaped cell bodies, much perinuclear cytoplasm and deep glial invaginations. Class-II cells (L3 and L4) have smaller perikarya (4–8 m) with little perinuclear cytoplasm and no glial invaginations. The midget monopolar cell (L5) resides at the base of the cell-body layer and has a cubshaped cell body. Though embedded within a reticulum of satellite glia, the L1–L4 monopolar perikarya and their immediately proximal neurites frequently appose each other directly. Typical arthropod (-type) gap junctions are routinely observed at these interfaces. These junctions can span up to 0.8 m with an intercellular space of 2–4 nm. The surrounding nonspecialized interspace is 12–20 nm. Freezefracture replicas of monopolar appositions confirm the presence of -type gap junctions, i.e., circular plaques (0.15–0.7 m diam.) of large (10–15 nm) E-face particles. Gap junctions are present between Class I somata and their proximal neurites, between Class I and Class II somata and proximal neurites, and between Class II somata. Intercartridge coupling may exist between such monopolar somata. The cell body and proximal neurite of L5 were not examined. We also find that Class I and Class II somata are extensively linked to their satellite glia via gap junctions. The gap width and nonjunctional interspace between neuron and glia are the same as those found between neurons. The particular arrangement and morphology of lamina monopolar neurons suggest that coupling or low resistance pathways between functionally distinct neurons and between neuron and glia are probably related to the metabolic requirements of the nuclear layer and may play a role in wide field signal averaging and light adaptation.     

Lateral connections between heart muscle cells as revealed by conventional and high voltage transmission electron microscopy

Summary The lateral surfaces of heart muscle cells are interconnected by a varied and extensive network of structures that exist in addition to intercalated discs. Ultrastructural images of this network are vastly improved over those from epoxy-embedded material, particularly for low density components, through the application of a method for removing the embedding matrix from thin or thick sections that are then stereoscopically analyzed with standard or high voltage transmission electron microscopy. The connections include cables, 3–20 nm in diameter, multi-strand cables, 10–40 nm-granules, meshlike mats, and sheets, all extensively interwoven. It is suggested that intercellular connections of varying strength and distribution aid in the integration of mechanical performance of the large population of myocytes during the contractile cycle of the heart.This study was supported by a grant from NIH Biotechnology Resources through the University of Colorado High Voltage E.M. Laboratory, NIH Research Grant HL 24336, a N.Y. Heart Association Grant-in-Aid, and NIH Research Career Development Award HL 00568I thank Dr. E.H. Sonnenblick for continual aid and encouragement and Dr. R. Terry, Ms. Y. Kress, and Ms. J. Fant for use of facilities. I also thank Dr. K.R. Porter for guidance in the use of the HVEM technique, Dr. J.J. Wolosewick and Dr. M. Fotino for valuable suggestions, and Ms. J. Fleming, Mr. G. Wray, and Mr. G. Charlie of HVEM staff at Boulder. I acknowledge Dr. F. Pepe for use of facilities, Dr. R. Bloodgood for comments, and Mrs. L. Cohen-Gould, Ms. T. Downey, Mr. F. Reingold, Mrs. T. Maio, and Mrs. R. Shamoon for excellent assistance     

A Golgi-electron-microscopical study of the structure and development of the lamina ganglionaris of the locust optic lobe

Summary The gross structure as well as the neuronal and non-neuronal components of the lamina ganglionaris of the locust Schistocerca gregaria are described on the basis of light- and electron-microscopical preparations of Golgj (selective silver) and ordinary histological preparations. The array of optic cartridges within the lamina neuropile — their order and arrangement — and the composition of the cartridges are described. There are six types of monopolar neurons: three whose branches reach to other cartridges and three whose branches are confined to their own cartridges. Retinula axons terminate either in the lamina or the medulla neuropiles. There are three types of centrifugal neurons, two types of horizontal neuron, as well as glia and trachea in the lamina neuropile. The development of the lamina neuropile is described in terms of developing monopolar and centrifugal axons, growing retinula fibres, and composition of the developing optic cartridges.MSN was supported in part by a Fulbrights-Hays Scholarsship. We are grateful to the Science Research Council for its grant to PMJS.     

X-ray microanalysis of biological specimens by high voltage electron microscopy

For the purpose of analyzing and imaging chemical components of cells and tissues at the electron microscopic level, 3 fundamental methods are available, chemical, physical and biological. Among the physical methods, two methods qualifying and quantifying the elements in the structural components are very often employed. The first method is radioautography which can demonstrate the localization of radiolabeled compounds which were incorporated into cells and tissues after the administration of radiolabeled compounds. The second method is X-ray microanalysis which can qualitatively analyze and quantify the total amounts of elements present in cells and tissues. We have developed the two methodologies in combination with intermediate high or high voltage transmission electron microscopy (200–400 kV) and applied them to various kinds of organic and inorganic compounds present in biological materials. As for the first method, radioautography, I had already contributed a chapter to PHC (37/2). To the contrary, this review deals with another method, X-ray microanalysis, using semi-thin sections and intermediate high voltage electron microscopy developed in our laboratory.

X-ray microanalysis is a useful method to qualify and quantify basic elements in biological specimens. We first quantified the end-products of histochemical reactions such as Ag in radioautographs, Ce in phosphatase reaction and Au in colloidal gold immunostaining using semithin sections and quantified the reaction products observing by intermediate high voltage transmission electron microscopy at accelerating voltages from 100 to 400 kV. The P/B ratios of all the end products Ag, Ce and Au increased with the increase of the accelerating voltages from 100 to 400 kV. Then we analyzed various trace elements such as Zn, Ca, S and Cl which originally existed in cytoplasmic matrix or cell organelles of various cells, or such elements as Al which was absorbed into cells and tissues after oral administration, using both conventional chemical fixation and cryo-fixation followed by cryo-sectioning and freeze-drying, or freeze-substitution and dry-sectioning, or freeze-drying and dry-sectioning producing semithin sections similarly to radioautography. As the results, some trace elements which originally existed in cytoplasmic matrix or cell organelles of various cells in different organs such as Zn, Ca, S and Cl, were effectively detected. Zn was demonstrated in Paneth cell granules of mouse intestines and its P/B ratios showed a peak at 300 kV. Ca was found in human ligaments and rat mast cells with a maximum of P/B ratios at 350 kV. S and Cl were detected in mouse colonic goblet cells with maxima of P/B ratios at 300 kV. On the other hand, some elements which were absorbed by experimental administration into various cells and tissues in various organs, such as Al in lysosomes of hepatocytes and uriniferous tubule cells in mice was detected with a maximum of P/B ratios at 300 kV.

From the results, it was shown that X-ray microanalysis using semi-thin sections observed by intermediate high voltage transmission electron microscopy at 300–400 kV was very useful resulting in high P/B ratios for quantifying some trace elements in biological specimens. These methodologies should be utilized in microanalysis of various compounds and elements in various cells and tissues in various organs.     

Low and high voltage electron microscopy of mitosis and cytokinesis in maize roots

The structure and distribution of cytoplasmic membranes during mitosis and cytokinesis in maize root tip meristematic cells was investigated by low and high voltage electron microscopy. The electron opacity of the nuclear envelope and endoplasmic reticulum (ER) was enhanced by staining the tissue in a mixture of zinc iodide and osmium tetroxide. Thin sections show the nuclear envelope to disassemble at prophase and become indistinguishable from the surrounding ER and polar aggregations of ER. In thick sections under the high voltage electron microscope the spindle is seen to be surrounded by a mass of tubular (TER) and cisternal (CER) endoplasmic reticulum derived from both the nuclear envelope and ER, which persists through metaphase and anaphase. At anaphase strands of TER traverse the spindle between the arms of the chromosomes. The octagonal nuclear pore complexes disappear by metaphase, but irregular-shaped pores persist in the membranes during mitosis. It is suggested that these form a template for pore-complex reformation during telophase. Phragmoplast formation is preceded by an aggregation of TER across the spindle at anaphase. Evidence is presented to suggest that the formation of the desmotubule of a plasmodesma is by the squeezing of a strand of endoplasmic reticulum between the vesicles of the cell plate.Abbreviations CER cisternal endoplasmic reticulum - ER endoplasmic reticulum - HVEM high voltage electron microscope - TER tubular endoplasmic reticulum - ZIO zinc iodide/osmium tetroxide     

Direct connections between mitochondria and catecholamine-storage vesicles demonstrated by high voltage electron microscopy in rat adrenal medulla

Summary Tubular channels from mitochondria to catecholamine-storage vesicles have been demonstrated in thick sections of adrenal medullary tissue from hypoglycemia-stressed rats by the use of the high voltage electron microscope. The function of these connections is not presently known although they may serve as channels for the transport of materials such as high-energy nucleotides from one organelle to the other. The present study has examined only the adrenal medulla, but it should be considered that such connections may also exist in other neural cells and possibly other cells in which there is intracellular transport of ATP.This work was supported by NIH DRR 70-4136, WVU Medical Corporation Research Grants, Biomedical Research Support Grant 5 SOI RR05433, WVU Senate Research Grant, and West Virginia Heart Association Research Grant. We wish to thank Barbara Coalgate, Billie Pack, and the staffs of the HVEM laboratories at U.S. Steel and University of Colorado.     

Evaluation of commercial and field‐expedient baited traps for house flies,Musca domestica L. (Diptera: Muscidae)

A comparison of nine commercial baited fly traps on Florida dairy farms demonstrated that Terminator traps collected significantly more (13,323/trap) house flies (Musca domestica L.) than the others tested. Final Flight, Fly Magnet, and FliesBeGone traps collected intermediate numbers of flies (834‐2,166), and relatively few were caught with ISCA, Advantage, Fermone Big Boy, Squeeze & Snap, or OakStump traps (<300). Terminator traps collected about twice as many flies (799.8/trap) as FliesBeGone traps (343.8) when each trap was baited with its respective attractant, but when the attractants were switched between the two trap types, collections were significantly lower (77‐108) than was observed with traps baited with their respective attractant. Solutions of molasses were significantly more attractive to house flies than honey, maple syrup, or jaggery (date palm sugar). Field‐expedient traps constructed from discarded PET water bottles were much less effective than commercial traps, but painting the tops of such traps with black spray paint resulted in a six‐fold increase in trap capture.     

Imaging of plant microtubules with high resolution scanning electron microscopy

Summary High resolution scanning electron microscopy was used to obtain images of cortical microtubules and associated structures in onion root tips. Specimens were prepared using a modified quick-freeze deep-etch technique utilising cytosolic extraction with saponin and conductive staining with osmium.Abbreviations DMSO dimethylsulfoxide - HRSEM high resolution scanning electron microscope/microscopy - MTSB microtubule stabilising buffer - TEM transmission electron microscope/microscopy     

Genetic control of LDH isozymes in the house fly,Musca domestica

Electrophoretic variations in lactate dehydrogenase from adult whole body homogenates are described for three laboratory strains of house fly, Musca domestica. Several crosses between different electrophoretic forms provided evidence that the observed variations are due to segregation of alleles at two distinct loci (designated as A and B loci) and that the LDH isozymes of house flies are dimers formed by a random association of subunits controlled by the two loci.     

The superposition eye of Cloeon dipterum: The organization of the lamina ganglionaris

Summary The lamina ganglionaris of the superposition eye of Cloeon dipterum is composed of separate optic cartridges arranged in a hexagonal pattern. Each optic cartridge consists of one central, radially branched monopolar cell (Li) surrounded by a crown of seven retinula cell terminals and two more unilaterally branched monopolar cells (La1/La2) situated close together outside the cartridge. Projections to neighbouring cartridges have not been observed.In most cases, synaptic contacts could be seen between a presynaptic retinula cell and more than two other postsynaptic profiles, which belong to monopolar cells or sometimes to glial cells.Seven retinula cell fibers of one ommatidium pass in a bundle through the basement membrane, run into their respective cartridges without changing orientation and terminate at approximately equal levels in the lamina. Long visual fibers with endings in the medulla are not visible in the superposition eye lamina, but are present in the lateral apposition eye. The relationship between the behaviour of the animal, optic mechanisms of the superposition eye and the structure of the lamina is discussed.     

Kinetic characteristics of native gamma-glutamylcysteine ligase in the aging housefly, Musca domestica L

The catalytic activity of gamma-glutamylcysteine ligase (gamma-GCL; EC 6.3.2.2) was compared between relatively young (4-day-old) and old (19-day-old) houseflies (Musca domestica) in order to understand the mechanism of putative deterioration of glutathione homeostasis during the aging process. Hanes-Woolf analyses ([S]/v vs [S]) indicated that gamma-GCL had significantly higher affinities for its substrates in the young than in the old flies. The K(m) values in the young and old flies were, respectively, for glutamate 0.6 and 5.5 mM; for cysteine 0.3 and 4.6 mM; and for ATP 1.2 and 2.9 mM. Furthermore, young but not old flies exhibited substrate-dependent inhibition of gamma-GCL activity at >5 mM cysteine indicating a loss of metabolic regulation during aging. The age-associated differences in the affinity of native gamma-GCL towards its substrates suggest that de novo synthesis of glutathione would be relatively less efficient in the old houseflies.     

Degradation of cuticle during larval-pupal and pupal-adult development of the housefly, Musca domestica

Abstract. The patterns of changes in cuticle weight, its chitin content and chitinase activity have been studied during postembryonic development of the housefly, Musca domestica L. During pupariation the larval cuticle loses weight. During the early part of this weight-loss the decline in chitin content parallels the overall change in cuticle weight. A simultaneous elevation in chitinase activity suggests that at this time the larval cuticle is being enzymatically degraded. Later weight loss may be due to sclerotization. No significant changes in cuticle weight or its chitin content occur in pharate cuticle until one day before eclosion. However, a peak of chitinase activity found at mid-late pupal stage suggests the timing of pupal cuticle breakdown.     

Evaluation of (Z)-9-tricosene baited targets for control of the housefly (Musca domestica) in outdoor situations

Houseflies (Musca domestica L.) are a major pest species in a variety of outdoor situations, notably on and around livestock farms and landfill used for the disposal of domestic waste. Currently no effective options are available for control of houseflies outdoors, because many populations exhibit at least some resistance to all available synthetic pesticides. (Z)‐9‐tricosene is the only commercially available pheromone for use in lure‐and‐kill approaches to housefly control, and it is widely used in indoor livestock units in combination with sugar/insecticide bait. Here we examine the potential of this approach for use outdoors, on a landfill site. We investigate the efficacy of toxic targets painted with a sugar/insecticide/(Z)‐9‐tricosene mix. The effects of target size and pheromone concentration were examined in two replicate trials, conducted in June and September 2003. As expected, catch consisted largely of males, was consistently higher on larger traps, and generally increased with (Z)‐9‐tricosene concentration even up to very high levels. However, in repeated trials, and despite mass release of marked flies, catch rates appeared to be insufficient to provide adequate control. We suggest that this is probably because (Z)‐9‐tricosene is primarily a short‐range attractant, and fly populations in outdoor situations are generally distributed over a large area. Catch declined rapidly to zero within 2 weeks, indicating that improved formulation or target design is needed to slow the weathering of active ingredients on the targets. It seems unlikely that (Z)‐9‐tricosene is sufficiently attractive to houseflies to provide an effective and economic lure in outdoor situations.