Intranuclear dynamics of chromosome 6 in nurse cells of Calliphora erythrocephala Mg. (Diptera: Calliphoridae)

Intranuclear dynamics of chromosome 6 in nurse cell nuclei of Calliphora erythrocephala Mg. (Diptera: Calliphoridae) was studied. The 3D FISH method was used for the first time to study chromosome territories in highly polyploid nuclei whose chromosomes undergo morphological changes. A considerable change in the intranuclear location of chromosome 6 and a morphological alteration of the chromosome territory in the course of chromatin polytenization were revealed.     

Visualization of chromosome territories in interphase nuclei of ovarian nurse cells in Calliphora erythrocephala Mg. (Diptera: Calliphoridae)

Analysis of localization of chromosomes 2, 3, and 6 of Calliphora erythrocephala Mg. in ovarian nurse cell nuclei with different chromatin structure has shown that the regions of DNA probe hybridization reduced with increasing chromatin compaction. Hybridization of DNA probes of chromosomes 3 and 6 to secondary reticular nuclei demonstrated that chromosomes retain their territories in the nuclei when the chromatin acquires a reticular structure. These results suggest regular organization of the chromosomal apparatus at all stages of the endomitotic cycle, including the stage of highly polyploid reticular nuclei. FISH of DNA probe of the chromosome 2 telomeric region to secondary reticular nuclei revealed a peripheral distribution of the signal. Zones of more intensive DNA probe hybridization have been distinguished. These zones probably are the regions of accumulation of telomeric and (or) centromeric chromosome regions.     

Cyst geometry in the egg chambers of Calliphora erythrocephala Mg. (Diptera: Calliphoridae) ovaries

In the germarium of polytrophic ovarioles of Calliphora erythrocephala (Mg.) fly, four mitotic divisions of cystoblasts give rise to 16-cell germ-line cysts. One cell differentiates into an oocyte, while the remaining 15 cells become nurse cells. Concomitantly actin-rich ring canals are formed at the intercellular junctions. The present study considers a mutual arrangement of the ring canals formed after the second to fourth mitoses relative to the ring canal formed after the first mitotic division in different regions of the germarium and egg chambers. During the cyst formation and its movement to the posterior end of the germarium, the ring canals are displaced relative to one another, thereby giving different branching variants of the cyst. The pattern of cell interconnections becomes stable in germarium region 2b and does not change during the cyst movement along the ovariole despite the cyst polarizes and increases in size.     

Intranuclear dynamics of chromosome 6 in nurse cells of <Emphasis Type="Italic">Calliphora erythrocephala</Emphasis> Mg. (Diptera: Calliphoridae)

Intranuclear dynamics of chromosome 6 in nurse cell nuclei of Calliphora erythrocephala Mg. (Diptera: Calliphoridae) was studied. The 3D FISH method was used for the first time to study chromosome territories in highly polyploid nuclei whose chromosomes undergo morphological changes. A considerable change in the intranuclear location of chromosome 6 and a morphological alteration of the chromosome territory in the course of chromatin polytenization were revealed.     

The haemocytes of Calliphora erythrocephala (Meig.) (Diptera)

Summary The haemocytes of larvae and young pupae of Calliphora erythrocephala are studied by phase contrast and electron microscopy and three cell lineages are distinguished: plasmatocytes, thrombocytoids and oenocytoids. The plasmatocytes show important modifications during larval development and at the time of histolysis, which are described and discussed in relation to the function of these cells in the physiology of Calliphora. The thrombocytoids, haemocytes which had not been recorded so far, are characterized by a strong tendency to fragmentation, this process leading to the formation of the anucleated cytoplasmic fragments and the naked nuclei referred to by earlier authors. The ability of the cell fragments, which retain normal cytological characteristics, to agglutinate and form intricate networks, is discussed in relation to haemostasis in Calliphora.The ultrastructural study of the haemocyte accumulations in the vicinity of the posterior part of the dorsal vessel reveals the basic organization of haemocytopoetic tissue, as described recently in orthopteran insects. The functional importance of this tissue in the production of haemocytes is demonstrated by X-irradiation and ligation experiments in larvae of Calliphora.

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Résumé L'étude en microscopie en contraste de phase et au microscope électronique permet de distinguer dans le sang circulant de larves et de jeunes pupes de Calliphora erythrocephala trois lignées cellulaires: les plasmatocytes, les thrombocytoïdes et les oenocytoïdes. Les plasmatocytes, numériquement les plus importants, présentent au cours du développement larvaire et chez les pupes des modifications considérables, qui sont décrites et discutées en rapport avec la fonction évidente de ces hémocytes chez Calliphora. Les thrombocytoïdes, inconnus dans la littérature, se caractérisent par une forte tendance à la fragmentation, qui aboutit à la formation des «fragments cytoplasmiques anucléés» et des «noyaux nus» signalés par divers auteurs. Les phénomènes d'agglutination des «fragments cytoplasmiques anucléés», dont les caractères cytologiques restent normaux, sont discutés en rapport avec le problème de l'hémostase chez cet insecte.L'étude ultrastructurale des accumulations hémocytaires autour du vaisseau dorsal dans la partie postérieure de l'abdomen montre une organisation de base comparable à celle décrite dans les organes hématopoïétiques des Insectes Orthoptères. L'importance fonctionelle de ce tissu hématopoïétique de Calliphora dans la production des hémocytes au cours de la vie larvaire est démontrée par des irradiations de ce tissu et par des ligatures de la partie postérieure de l'abdomen.

     

Organization and differential staining of chromosomes from the endomitotic nucleus of trophocytes Calliphora erythrocephala (Diptera: Calliphoridae)

The structure of primary polytene chromosomes and general architecture of nurse cell nuclei was studied in Calliphora erythrocephala using various methods of differential chromosome banding(G-, R-, C-banding; Ag- and DAPI staining), chromospecific DNA probes and fluorescence in situ hybridization. This analysis revealed differential compaction of particular chromosome regions. The localization of material of polytene chromosome 6 is retained after its rearrangement and the formation of the internal reticular structure of the nucleus. Polytene chromosomes of ovarian nurse cells were shown to have blocks of dense compact material; some of them were more intensely stained by AgNO3. The dynamics of the nucleolus formation was traces at all stages of chromosome polytenization in the C. erythrocephala nurse cells.     

Chromosome Organization and Differential Banding in Endomitotic Nuclei of Nurse Cells of Calliphora erythrocephala (Diptera: Calliphoridae)

The structure of primary polytene chromosomes and general architecture of nurse cell nuclei was studied in Calliphora erythrocephala using various methods of differential chromosome banding(G-, R-, C-banding; Ag-, and DAPI staining), chromospecific DNA probes and fluorescence in situ hybridization. This analysis revealed differential compaction of particular chromosome regions. The localization of material of polytene chromosome 6 is retained after its rearrangement and the formation of the internal reticular structure of the nucleus. Polytene chromosomes of ovarian nurse cells were shown to have blocks of dense compact material; some of them were more intensely stained by AgNO₃. The dynamics of the nucleolus formation was traces at all stages of chromosome polytenization in the C. erythrocephala nurse cells.     

A quantitative analysis of mitotic gradients during early development in Calliphora erythrocephala Meig. (Diptera)

Mitotic waves during superficial cleavage and early gastrulation were analyzed quantitatively in Calliphora. Three consecutive patterns are present: (1) a monotonic anterioposterior mitotic gradient during early superficial cleavage; (2) a double mitotic gradient from the anterior and posterior poles during superficial cleavage, especially toward the end of the period; and (3) more complicated patterns with intermediate mitotic centers during the last superficial cleavage division and during early gastrulation. Mitotic gradients are absent in many eggs during early superficial cleavage, but they then become ubiquitous. The gradients are longitudinal; no transverse component was detected before gastrulation. Anterior and posterior gradient patterns are not mirror images of each other; mitotic activity always starts earlier anteriorly. The gradients are accompanied by a pronounced increase in interphase length. The mitotic gradients are compared with the morphogenetic gradients predicted in a current model for pattern specification in insect eggs.     

Mechanisms of wing rotating regulation in Calliphora erythrocephala (Insecta,Diptera)

Summary The blowfly Calliphora erythrocephala rotates its wings, i.e. changes the geometrical angle of attack, generating forces and moments for flight steering. There are two possibile ways to regulate this angle. The mechanisms for these movements are described. (1) The leading edge and the anterior part of the wing — between the costal vein and radial vein 4 — are pronated automatically due to the interaction of the moving parts during the downstroke. They are supinated during the upstroke. This is basic automatic regulation. (2) The posterior part of the wing — behind the anterior cross vein — is pronated and supinated independently of wing-drive. This is wing-drive independent additional regulation.Abbreviations a.c anterior cross vein - a.n anal veins - a.t.l anterior tergal lever - a.w anterior part of the wing - b.z bending zone - co costal vein - cr crossing of the tendons of the posterior notal wing process - c.s cross section - cu cubital vein - f fit or turning point of ventral radial vein 1 - h.a horizontal axis of pterale III - h.c humeral cross vein - h.co head of costal vein - h.r head of radial vein - k Klöppel - l.a longitudinal axis - me median vein - mp middle plate - ms mesoscutum - p anterior process of the anal veins - p.c posterior cross vein - pl pleurum - p.n.w.p posterior notal wing process - p.n.w.p 1–4 muscles 1–4 of the posterior notal wing process - pt I–III pterale I–III - p.t.l posterior tergal lever - p.w. posterior part of the wing - p.w.j pleural wing joint - r 1–4 radial veins 1–4 - r.s. ring stiffenings - sc subcostal vein - s.p semicircular part of the middle plate - s.t subalar tendon - t.c tip cross vein - te tegula - t.st thin strips - t.v.r tooth of ventral radial vein - v.a. vertical axis of pterale III - w wing - III 1–4 muscles 1–4 of pterale III     

Hybrid carrot seed crop pollination by the fly Calliphora vicina (Diptera: Calliphoridae)

Many insect species can contribute to crop pollination; however, most growers remain highly dependent on the managed honey bee (Apis mellifera L.) for this service. The European Blue Blow Fly Calliphora vicina Robineau‐Desvoidy, 1830 is one species with potential use as a pollinator. It occurs worldwide and is easy to rear. Caged trials conducted within a hybrid carrot (Daucus carota L.) seed crop found C. vicina to be an effective pollinator. Seed yield (number and weight) from field‐grown carrot plants caged with C. vicina, but excluding all other large flower visitors (body width > 3 mm), was similar to seed yield from uncaged plants in the presence of honey bees (Apis mellifera L.) and other insects. In contrast, caged plants without C. vicina produced 10‐fold less seed. Under open field conditions, C. vicina spent an average of 71.0 s per umbel compared to 54.4 s for honey bees; however, under caged conditions, C. vicina spent more time on average per umbel (128.9 s). Counts of C. vicina and honey bees on umbels outside of cages and C. vicina inside cages found that honey bees were most abundant on days with maximum temperature > 25°C, while C. vicina was more abundant on cooler days around 20°C. C. vicina may therefore be a useful pollinator of crops grown in isolation cages for plant breeding purposes as well as in open fields when climatic conditions are less favourable for optimal honey bee activity.     

Endopolyploidy in follicle epithelial cells around the nurse chamber during oogenesis in Chrysomya putoria (Diptera,Calliphoridae)

Feulgen-DNA values and nuclear areas were evaluated microspectrophotometrically for epithelial cells of the ovarian follicle during oogenesis in Chrysomya putoria. The aim was to investigate whether polyploidization occurred in the cells surrounding the nurse chamber and/or in those around the oocyte as well as whether different DNA amounts were found regarding the cell types considered. Four DNA endoreduplicative cycles could be demonstrated for the epithelial cells regardless of their localization on the follicle, during oogenesis. A small percentage of epithelial cells reaches a 32C degree. The nuclear area, however, did not increase at the same rate in cells covering the oocyte as in those covering the nurse chamber, in some of the oogenesis stages. The meaning of endopolyploidy for these cells is discussed, considering reports on relevance of secretory activities and their maintenance in polyploid cell systems.     

The contribution of the fatbody to RNA and ribosomal changes during development of the blowfly Calliphora erythrocephala (Meig)

The initiation of blowfly metamorphosis is associated with a pronounced decrease in the number of larval ribosomes; this reduced number then remains constant throughout pharate adult development. Ribosomal RNA accounts for most of the total RNA in larvae shortly after the cessation of feeding and growth, but thereafter the amount of rRNA declines disproportionately to total RNA until early pharate adult development; thereafter, the ratio remains constant until adult emergence. Larval fatbody ribosomes, which constitute about half of the total in the entire organism, are destroyed in situ prior to pupariation. The progressive decrease in fatbody rRNA is accompanied by a corresponding increase in a degraded, relatively insoluble 4–7 S nucleic acid which, stored until adult emergence when it is discarded, accounts for the disparity between total RNA and rRNA. The extracellular ribosomes previously observed during pharate adult development are thus derived from dissolution of larval tissues other than fatbody.     

Ultrastructure of the myocardial cell and its membrane systems in the adult fly Calliphora erythrocephala (Insecta: Diptera)

Summary Calliphora erythrocephala has cross-striated cardiac muscle cells with A, I and Z-bands. The diameters of the myosin and actin filaments are 200–250 Å and 85 Å respectively and the length of the myosin filaments (A-band) is approximately 1.5 . Usually 8–10 actin filaments surround each myosin filament.The myocardial cells show a well-developed membrane system and interior couplings. A perforated sheet of SR envelopes the myofibrils at the A-band, dilates into flattened cisternae at both A-I band levels before it merges into a three-dimensional net-work between the actin filaments of the I-bands and between the dense bodies of the discontinuous Z-discs. The T-system consists of broad flattened tubules running between the myofibrils at the A-I band levels forming dyads with the SR-cisternae. Longitudinal connections between the transverse (T-) tubules often occur.It is suggested that this well-developed SR may be an adaptation to facilitate a rapid contraction/relaxation frequency by an effective Ca²⁺ uptake.     

Ultrastructure and ontogeny of the hair sensilla on the funicle of Calliphora erythrocephala (Insecta,Diptera)

Summary Four envelope cells are responsible for the formation of the basiconical sensilla of Calliphora. They are the thecogen, trichogen, and tormogen cells, and envelope cell 4. In early stages of development the still subepithelial sensory cilia are completely enclosed by the innermost thecogen cell. The first formation movements are initiated by a growth thrust of the hair-forming cell into the exuvial space. The sensory cilia only begin to grow into the hair anlage when the hair-forming cell has almost reached its final length. As soon as growth is completed the trichogen cell, tormogen cell, and envelope cell 4 start to excrete cuticular material. The trichogen cell forms the perforated part of the hair shaft and the stimulus-conducting system consisting of the pore tubules. The tormogen cell is responsible for the excretion of the basal non-perforated hair shaft and sheath cell 4 forms the proximal part of the socket region. The thecogen cell only begin to produce dendritic sheath material when the sensory hair is almost complete.Approximately 7–8 days after pupation the tormogen cell degenerates, having, by this time, produced about two-thirds of the sensilla cuticle. The surrounding envelope cells incorporate cell fragments of the tormogen cell. The trichogen cell continues the secretion where the tormogen cell left off. When the secretion of cuticle is finished the sheath cells begin to withdraw towards the proximal direction and to form microvilli on the apical membrane. The resulting outer receptor lymph space is bordered by envelope cell 4 and the trichogen and thecogen cells. The tormogen cell is absent in the sensilla of the imago.Abbreviations DS dendritic sheath - E4 envelope cell 4 - Ex exuvial space - G glial cell - iD inner dendritic segment - iRL inner receptor lymph space - oRL outer receptor lymph space - oD outer dendritic segment - P pore - PT pore tubules - S sensory cell - T thecogen cell - TO tormogen cell - TR trichogen cell Part 1 of a dissertation accepted by the Faculty of Bio- and Geosciences, University of Karlsruhe     

Contact behaviour of tenent setae in attachment pads of the blowfly Calliphora vicina (Diptera, Calliphoridae)

To enable strong attachment forces between pad and substrata, a high proximity between contacting surfaces is required. One of the mechanisms, which can provide an intimate contact of solids, is a high flexibility of both materials. It has been previously presumed that setae of hairy attachment pads of insects are composed of flexible cuticle, and are able to replicate the surface profile. The aim of this work was to visualise the contact behaviour of the setae by freezing-substitution technique to understand setal mechanics while adhering to a smooth surface. This approach revealed considerable differences in the area of the setal tips between contacting and non-contacting pulvilli. Based on the assumption that setae behave like a spring pushed by the tip, a spring constant of 1.31 N m(-1) was calculated from direct measurements of single setae by atomic force microscopy. In order to explain the relationship between the behaviour of the attachment setae at a microscale and leg movements, high-speed video recordings were made of walking flies. This data show that some proximal movement of the leg is present during contact formation with the substrate.