Ecdysone Oxidase, an enzyme from the blowfly Calliphora erythrocephala (Meigen).

In the blowfly, the formation of 3-dehydroecdysone from the insect molting hormone ecdysone is catalyzed by an enzyme which carries hydrogen from ecdysone and ecdysterone to oxygen. The enzyme is therefore called "ecdysone oxidase". Two methods are described for the detection of ecdysone oxidase activity, one using a radiolabelled substrate which is separated from the product by thin-layer chromatography after the reaction, and the other using dichloroindophenol, which is discoloured by the redox reaction. The ecdysone oxidase is purified by a factor of 2200 from prepupae of Calliphora erythrocephala using salt precipitation and ion exchange chromatography. The ecdysone oxidase has a Km value for ecdysone of 42muM. The pH optimum is 6.5. The temperature optimum lies at 45 degrees C. The ecdysone oxidase has a molecular weight of 240000.     

Salivary gland development in the blowfly,Calliphora erythrocephala

The salivary gland of adult Calliphora erythrocephala is a tubular structure composed of secretory, reabsorptive, and duct regions. Development of these structures has been followed during the six days of larval and ten days of pupal growth. Two small groups of imaginal cells located at the junction between larval gland and duct give rise to the adult gland. These presumptive adult cells divide during all larval stages and appear to be functional components of the larval gland. Shortly after pupation, the larval gland breaks down and the imaginal cells proliferate rapidly, forming sequentially the duct, reabsorptive and secretory regions. Proliferating regions of the developing gland are frequently encrusted with haemocytes. As it elongates the gland establishes intimate contacts first with the basement membrane of the degenerating larval gland, later with an epithelial layer surrounding the main dorsal tracheal trunks, and then with the gut. Cell division continues until about five days after pupation, bu t the gland is unable to secrete fluid in response to 5-hydroxytryptamine stimulation until two hours after the adult fly emerges. The Golgi complex appears to be involved in forming the highly folded membranes of the canaliculi in the secretory region. Presumptive adult salivary gland cells appear to increase in number logarithmically from the time of hatching of the larva until five days after pupation. This contrasts with the development of classical imaginal discs, in which cell division ceases prior to pupation.     

On the landing response of the blowfly,Calliphora erythrocephala

The dependence of the landing response on the direction of moving stimuli (periodic gratings, single or double stripes) was studied in blowflies, Calliphora erythrocephala, of both sexes. Directions of motion eliciting maximally strong responses (preference direction) vary with the eye region stimulated: they are distributed radially from a common origin forming a flow-field. This origin lies at the intersection of the eye equators with the median plane of the animal. By changing its body posture relative to the direction of flight, the fly may align the pole of this flow field-with its direction of flight thus maximizing signal flow for the landing approach. Sex-specific differences were found for dorsal eye regions in which the shift of preference directions from vertical to obliquely inclined directions of motion (against the median plane) could only be determined for male flies.     

Hormonal control of resilin deposition in the blowfly, Calliphora erythrocephala

The deposition of the resilin tendon in the blowfly Calliphora erythrocephala was investigated in normal and in various experimental conditions. The results showed that the weight of the protein resilin that is deposited is controlled by diet as well as by the hormone secreted by the medial neurosecretory cells.Endocrinologically abnormal Calliphora adults deposited a tendon with normal ultrastructure but showed signs of premature ageing while Calliphora fed on a sugar diet deposited a tendon with abnormal ultrastructure.     

Elektrophysiologische Untersuchungen am Chemorezeptor von Calliphora erythrocephala Meigen

Zusammenfassung Bei Reizung des Chemorezeptors von Calliphora mit NaCl werden 2 Rezeptoren erregt: ein schnell adaptierender (L) und ein langsam adaptierender (M). Mit steigender Konzentration wird die Frequenz des L-Rezeptors erniedrigt und die des M-Rezeptors erhöht. Bei zusätzlicher Reizung mit Zucker antwortet der von Phormia her bekannte S-Rezeptor.Die Alkaloidglykoside Tomatin und Solanin rufen in Konzentrationen, die bei Leptinotarsa weit über dem Schwellenwert für den Einsatz einer Salve liegen, bei Calliphora lockere Salven vom S-Rezeptor hervor. Bei Zusatz von NaCl treten Salven mit hoher Frequenz auf, die vom M-Rezeptor und vermutlich auch S-Rezeptor stammen. Zusätzliche Reizung mit Zucker reduziert die Salven des M-Rezeptors augenblicklich zu Impulsgruppen, während der S-Rezeptor nach kurzer Zeit mit fortlaufender Erregung antwortet. Der L-Rezeptor wird durch die Alkaloidglykoside anscheinend nicht beeinflußt.     

Extracellular ribosomes during metamorphosis in the blowfly Calliphora erythrocephala

During metamorphosis of the blowfly Calliphora erythrocephala extracellular ribosomes, in the form of monosomes, appear in the body fluid. The total number of ribosomes, i.e. intracellular and extracellular, remains approximately constant during this period, whereas the proportion of extracellular ribosomes first rises, plateaus, and then declines in a manner suggesting that their appearance is a result of larval tissue breakdown.     

Cholinergic neurons in the lamina ganglionaris of the blowfly Calliphora erythrocephala

Summary The distribution of putative cholinergic neurons in the lamina of the blowfly Calliphora erythrocephala was studied by immunocytochemical and histochemical methods. Three different antibodies directed against the AChsynthesizing enzyme, choline acetyltransferase (ChAT), revealed a cholinergic population of fibres running parallel to the laminar cartridges, which have branch-like structures at the distal lamina border. Cell bodies in the chiasma next to the lamina border were also labelled by the anti-ChAT antibodies. Monopolar cell bodies in the nuclear layer were faintly labelled. The distribution of the acetylcholine hydrolyzing enzyme, acetylcholine esterase (AChE), was revealed by histochemical staining and was similar to the ChAT immunocytochemistry. The arrangement of ChAT positive fibres in transverse and longitudinal sections and the distribution of AChE stained fibres indicate that the amacrine cells of the lamina are cholinergic cells.We dedicate this work to Prof. F. Zettler who passed away in fall 1988: K.-H. Datum, I. Rambold     

Changes in acid mucopolysaccharides during the development of the blowfly, Calliphora erythrocephala.

Acid mucopolysaccharides have been isolated from different developmental stages of Calliphora. Hyaluronic acid and a ‘larval AMPS’ were identified during all developmental stages. During the later part of the development chondroitin and a poorly-sulphated keratin sulphate-like compound were also present. Chondroitin sulphate and heparan sulphate could be detected at all development stages and during the latter part possibly keratin sulphate. The variation of acid mucopolysaccharides during development is discussed.     

In vitro synthesis of peritrophic membranes of the blowfly, Calliphora erythrocephala.

Tyrode solution containing added glutamine and Leloup's medium 1 has been used as a basic medium for the in vitro culture of the so-called proventriculus of adult Calliphora erythrocephala to elucidate some of the factors controlling the synthesis of peritrophic membranes (PM) in vitro. The formation rate was chosen as a quantitative criterion for the evaluation of the modifications of the incubation media.After systematic variation of osmolarity, pH, and temperature optimal formation rates were obtained in media with an osmolarity of 320 to 360 mOsmol, a pH of 6·8, and an incubation temperature of 27°C. Under these conditions the average rate of formation was in the modified Tyrode solution 3·0±1·1 mm PM/hr, and in Leloup's medium 3·6±0·8 mm PM/hr. In the modified Tyrode solution the formation of PM was complete after 5 to 7 hr, whereas in Leloup's medium it continued up to 24 hr. The addition of β-ecdysone caused an increase of the formation rate of PM to 4·5 to 5·5 mm PM/hr.The results obtained led to the hypothesis that an osmotically regulated enzyme system could be the limiting factor of the formation rate of peritrophic membranes, i.e. a system which could regulate the internal osmolarity of the formative cells by the interconversion of a bulk polymer and its monomer which are needed for the synthesis of PM.     

Ocellar interneurones in the blowfly Calliphora erythrocephala: Morphology and central projections

Summary The morphology and central projections of first-order ocellar interneurones were analysed in the blowfly, Calliphora erythrocephala after cobalt and horseradish-peroxidase labelling. Three classes of interneurones can be distinguished on the basis of axon diameters: large, medium and small neurones. In total there are 12 large, 10 medium and an unknown number of small interneurones. These interneurones connect the fused first-order ocellar neuropil (underlying the three ocelli) with various areas of the central nervous system. The large neurones terminate in three subregions of the posterior slope (ocellar foci); the medium neurones arborise in several regions of the lateral protocerebrum, in the posterior slope, the lobula, the ventral medulla, and in the pro- and mesothoracic ganglia. The thin fibers arborise in all the above regions (except in the thoracic ganglia), and in addition in the neuropil dorsal to the oesophagus and antero-ventral to posterior slope (tritocerebrum). The anatomy of the ocellar pathway in C. erythrocephala is compared with those in other studied insects. Possible interactions between ocellar interneurones and other pathways are discussed.