Activation of latent ribonuclease in the fat-body of fleshfly (Sarcophaga peregrina) larvae on pupation.

A crude extract of the fat-bodies of third-instar larvae of Sarcophaga peregrina (fleshfly) was found to contain latent RNAase (ribonuclease) consisting of RNAase and inhibitor protein that is sensitive to p-chloromercuribenzoic acid. The RNAase activity in the crude extract of fat-bodies became detectable with time after puparium formation, indicating that the inhibitor is selectively inactivated and RNAase is released from the RNAase-inhibitor complex during metamorphosis.     

Comparative studies on ecdysone metabolism between mature larvae and pharate pupae in the fleshfly,Sarcophaga peregrina

Metabolites of radioactive ecdysone or 20-hydroxyecdysone in larvae and pharate pupae of Sarcophaga peregrina were separated and identified by using thin-layer chromatography, high-performance liquid chromatography, and chemical methods. At the larval stage ecdysone was metabolized to biologically less active ecdysteroids predominantly through 20-hydroxyecydsone, at the pharate pupal stage, to other ecdysteroids which were tentatively identified as 26-hydroxyecdysone, 3-epi-26-hydroxyecdysone, and 3-epi-20,26-dihydroxyecdysone. Ecdysteroid acids were found in the polar metabolites during pharate pupal-pupal transformation, but scarcely detected in the larval metabolites. These acids were presumed to be ecdysonoic acid, 20-hydroxyecdysonoic acid, and their epimers. The conjugates of ecdysteroid that released the free ecdysteroids by enzymatic hydrolysis were produced more in larvae than in pupae, whereas the very polar ecdysteroids that were not affected by the enzyme were found more in pupae. Therefore, there are different metabolic pathways of ecdysone between these two successive developmental stages, and the alteration of the metabolic pathway may serve as one of the important factors in a regulatory mechanism of molting hormone activity which is responsible for normal development of this insect.     

Participation of hemocyte proteinase in dissociation of the fat body on pupation of Sarcophaga peregrina (flesh fly)

In holometabolous insects, larval tissues are degraded on pupation. Previously, we established an in vitro system in which Sarcophaga fat body dissociates in the presence of pupal hemocytes, mimicking the situation in vivo (Kutata et al., J. Insect Physiol.35, 559–565, 1989). In this paper, we showed that chymostatin and phenylmethylsulfonyl fluoride prevented dissociation of fat body in this system. Moreover, of the various proteinases tested, only chymotrypsin degraded the fat body in vitro. These results suggested that a chymotrypsin-like proteinase of hemocytes participates in dissociation of the fat body at the early pupal stage.     

Induction of pupal diapause and photoperiodic sensitivity during early development of Sarcophaga peregrina larvae

Effects of long day and short day treatments during the embryonic and larval stages on induction of pupal diapause were studied on a diapausing race of Sarcophaga peregrina. Two long day (15L 9D) cycles during 2 days before or after the larviposition completely stopped the induction of pupal diapause on larvae which grow in short day condition before and after the long day treatment. The sensitivity appeared to decrease during the early stage of the third instar and to increase again to some extent in the prepupal stage.     

Involvement of Sarcophaga lectin in the development of imaginal discs of Sarcophaga peregrina in an autocrine manner

The imaginal discs of Sarcophaga were found not to develop normally in the presence of galactose, a hapten sugar of Sarcophaga lectin, or anti-Sarcophaga lectin antibody. Wing and leg discs cultured with these substances became morphologically abnormal and no imaginal discs reached the stage of terminal differentiation, even in the presence of 20-hydroxyecdysone. The development of the imaginal discs was shown to be autonomously regulated in an autocrine manner by Sarcophaga lectin; namely Sarcophaga lectin was secreted by the imaginal discs in the presence of 20-hydroxyecdysone, and the stimulus of self-induced Sarcophaga lectin seemed to be indispensable for further development of the imaginal discs. Sarcophaga lectin was originally found as a defense protein, but these results show that it plays independent roles in both defense and development.     

Identification of storage-protein messenger RNA of the fleshfly Sarcophaga peregrina.

Storage-protein mRNA was found to be abundant in poly(A)-containing RNA extracted from the fat-body of third-instar larvae of Sarcophaga peregrina (fleshfly). This RNA sedimented at the position of 19S on sucrose-density-gradient centrifugation and the product of its translation in vitro was 75K protein (protein of mol.wt. 75 000), which was precipitated specifically with antibody against storage protein. This product was suggested to contain a signal sequence that is missing in mature storage protein. The poly(A)-containing RNA was also found to contain much of another mRNA coding for 25K protein (protein of mol.wt. 25 000), but the function of this protein is unknown.     

Molecular cloning of cDNA for sapecin and unique expression of the sapecin gene during the development of Sarcophaga peregrina

A cDNA clone for sapecin, an antibacterial protein produced by an embryonic cell line of Sarcophaga peregrina, was isolated and characterized. This clone was found to encode a precursor of sapecin consisting of 94 residues, with sapecin (40 residues) constituting its carboxyl-terminal half. RNA blot hybridization revealed that the gene for the sapecin precursor is activated in the hemocytes of the third instar larvae of Sarcophaga in response to body injury. Thus, sapecin is probably a defense protein synthesized by Sarcophaga to prevent bacterial infection through the damaged body wall. This gene was also found to be activated in the embryonic and early pupal stages, suggesting that sapecin also plays a role in the ontogenetic processes of Sarcophaga.     

Changes in ecdysone titre during pupal-adult development in the silkworm, Bombyx mori

Changes in the ecdysone titre of the silkworm, Bombyx mori, during pupal-adult development were estimated. The average value of the maximum titre, which was observed on the second day after pupal ecdysis, was about 0·8 μg equivalent of β-ecdysone/g of live weight in both sexes.

There is a distinct sexual dimorphism in the pattern of the ecdysone titre. The male exhibited a single sharp peak on the second day whereas the female showed the second peak on the fifth day. When the female was ovariectomized, the ‘female type’ ecdysone pattern was converted to the ‘male type’. In the female pharate adult 7 days after pupal ecdysis, ecdysone activity accumulated in the ovaries.

The relationship between the ecdysone titre and adult differentiation, especially during ovarian development, is discussed.     

Purification of lectin induced in the hemolymph of Sarcophaga peregrina larvae on injury

A lectin was purified from the hemolymph of Sarcophaga peregrina larvae, obtained after injury of their body wall. This lectin agglutinated sheep red blood cells markedly and the hemagglutinating activity was inhibited by galactose and lactose. The active lectin was found to have a molecular weight of 190,000 and to consist of four alpha subunits and two beta subunits, with molecular weights of 32,000 and 30,000, respectively. During the early pupal stage, similar hemagglutinating activity in the hemolymph increased to several times than in larval hemolymph. This activity was completely inhibited by the antibody prepared against the lectin purified from the hemolymph of injured larvae. Thus, the same protein having lectin activity is apparently induced under two different physiological conditions: injury of the body wall of larvae and during pupation. The biological significance of this lectin is discussed.     

Determination and Regulation Capacities of the Wing Disc of the Fleshfly, Sarcophaga peregrina

Unilateral extirpation of the wing discs was performed on mid- and late-third instar larvae of Sarcophaga peregrina to investigate the regulation capacities of the thoracic discs. Out of 636 operated larvae, 175 became adult and all of them exhibited hemithoracic structures, lacking any adult structures in the operated side, which were normally produced from a wing disc. These findings suggest that the wing disc does not possess the capacity of bicentric regulation to produce the thoracic structures of the opposite side.     

Excretion of cadmium and change in the relative ratio of iso-cadmium-binding proteins during metamorphosis of fleshfly (Sarcophaga peregrina)

Cadmium (Cd) was accumulated in the larvae of Sarcophaga peregrina (fleshfly) by exposure through diet, and the metabolic fate of the accumulated Cd during metamorphosis was examined. Most of the Cd accumulated in the larvae was retained in the pupae, and 53% of that was excreted from the adults immediately after emergence. The Cd-binding protein induced in the larvae was degraded after pupation and the relative ratio of iso-Cd-binding proteins re-induced in the adults was different from that in the larvae. Copper content in the larvae was significantly reduced by Cd administration.     

Purification and Heterogeneous Localization of Sarcophaga Lectin Receptor on the Surface of Imaginal Discs of Sarcophaga peregrina (Flesh Fly)

Previously, we reported autocrine involvement of Sarcophaga lectin in the development of Sarcophaga imaginal discs (Kawaguchi et al. , Dev. Biol. 144 , 86–93 (1991)). In this study, we purified Sarcophaga lectin binding protein from the membrane fraction of cultured embryonic cells of Sarcophaga to near homogeneity and raised a monoclonal antibody against it. Histochemical analysis using the monoclonal antibody revealed that this binding protein is distributed heterogeneously on the surface of leg imaginal discs. This binding protein was especially clearly localized in the central region of the basal side of leg discs which forms the junction between the leg and body, suggesting the participation of Sarcophaga lectin in morphogenesis of the basal region of the developing leg.     

Visualizing the postembryonic development of Sarcophaga peregrina (flesh fly) by NMR microscopy

The postembryonic development of the flesh fly was studied using high resolution nuclear magnetic resonance imaging. Because this development occurs in a puparium, this process cannot be observed directly using standard histological techniques. The remodelling of histolysing larval tissues to developing imaginal tissues including the yellow body, a transient alimentary structure, and the integration of the developed adult structure were revealed in the images. Most surprisingly it was found that a large gas space that forms in the central region of the prepupa moves to the dorso-anterial region in less than 5 min due to the larval–pupal apolysis together with separation of the developing pupal epidermis from the puparium.     

Change in Free Amino Acids and Phospholipids in the Head of Adult Sarcophaga peregrina with Age

Abstract. The amino acid and phospholipid contents in the head of adult Sarcophaga peregrina (flesh-fly) were measured with age. Only aspartic acid was found to increase significantly with aging. A significant difference was also detected in the fatty acid composition of the phospholipids. Phospholipids isolated from old flies contained more unsaturated fatty acid than young flies, suggesting that the properties of the membrane of head cells change with age. These changes were observed irrespective of sex or rearing conditions. Thus, it is possible that certain metabolic pathways in post-mitotic cells are selectively affected by aging.