Vitellogenesis in the milkweed bug,Oncopeltus fasciatus Dallas (Hemiptera). A light and electron microscopic investigation

Histochemical and electron microscopic methods have revealed that there are four types of cell inclusions in the late vitellogenic oocytes of Oncopeltus. (a) Type 1 is a vacuole which seems to be contributed from the tropharium via the nutritive tubes. It is suggested that this type consists partly at least of nucleolus-like material (ribonucleoprotein) emitted from the nuclei of the Zone III trophocytes. (b) Type 2 is lipid yolk which in early stage oocytes seems to be produced in the “Balbiani body.” In the vitellogenic oocytes these lipid spheres are apparently imported by the oocyte from the haemolymph either through the follicle cells, or through the extracellular space in the follicular epithelium. (c) Type 3 is carbohydrate/protein yolk where at least part of the protein (“vitellogenic protein”) is taken up from the haemolymph, transported through the extracellular space in the follicular epithelium, and deposited into the oocyte by pinocytosis. (d) Glycogen is deposited from the early phases of vitellogenesis. The tropharium may contribute, besides Type 1 vacuoles, ribosomes, mitochondria, stacks of annulated lamellae, and “food vacuoles” to the oocytes. Specialized cells which line the tropharium and send projections toward the trophic core have been called “peripheral trophocytes.” Contrary to the regular trophocytes, they contain glycogen and an abundance of Golgi complexes.     

Carbohydrases in the alimentary canal of the large milkweed bug, Oncopeltus fasciatus (Hemiptera: Lygaeidae)

Adults of the large milkweed bug, oncopeltus fasciatus (Dallas), deprived of food for 48 hr, were fed 5% solutions of either sucrose, maltose, lactose, raffinose or melezitose, for 15–20 min. The alimentary canals were then dissected out 30 min or 1 hour after the period of feeding, and the gut contents chromatographed. The results indicated the presence in the gut of the following carbohydrases: -glucosidase, -fructosidase, -galactosidase and -galactosidase.

---

Résumé Des adultes de la punaise de l'asclépiade, Oncopeltus fasciatus (Dallas), soumis au jeûne pendant 48 heures, ont été nourris ensuite durant 15 à 20 minutes avec des solutions aqueuses de 5% de glucides, comprenant soit le sucrose le maltose, le lactose, le raffinose ou le mélézitose. Les tubes digestifs furent prélevés 30 ou 60 minutes après l'ingestion des sucres, et leur contenu glucidique séparé et identifié par chromatographie sur papier filtre. Les résultats ont révélé la présence dans le tube digestif des carbohydrases suivantes: -glucosidase, -fructosidase, -galactosidase et -galactosidase.

     

Acquired humoral immune response of the large milkweed bug,Oncopeltus fasciatus (Dallas), to injected materials

The large milkweed bug, Oncopeltus fasciatus (Dallas) responded to a single injection of Pseudomonas aeruginosa (Schroeter) Migula vaccine with the elaboration of lytic substances that were demonstrated in vitro in the serum. These factors appear to be responsible for the increased resistance to bacterial infection acquired by O. fasciatus in response to vaccination. Both the acquired resistance and lytic substances were detected 4 hr after vaccination. They reached their maximum level by 24 hr and disappeared within 5 days. A weak transitory resistance to Pseudomonas infection and low titre of lytic substances could be provoked by the injection of heterologous substances. Normal insects inoculated with immune serum exhibited a resistance equal to that acquired by individuals to specific vaccine.The acquired lytic substances resisted exposure to 75°C, 1 N HCl, and 1 N NaOH for 1 hr and retained their activity after storage at −20°C for at least 6 months. They are associated with several of the proteins in electrophoretically fractionated serum, to which they appear attached, but retain their activity in the supernatant of immune serum that has had the protein precipitated by trichloroacetic acid. They are not antigenic for rabbits, nor do they affect the antigenicity of the normal serum proteins.     

Regeneration of appendages in the large milkweed bug, Oncopeltus fasciatus

An analysis was made of the regeneration of legs and antennae of Oncopeltus. Amputations were performed on first instar larvae within 24 hr after hatching, and on later instars within 24 hr after ecdysis. The resulting regenerates were then measured at each instar. When amputations were performed soon after hatching, there was no significant effect on the duration of any instar. The regenerate was usually visible after the second post-operative ecdysis, and was smaller than a normal appendage (hypomorphic). Removal of the three distal segments of the antenna usually resulted in regeneration of only one segment which was abnormally long and showed a combination of the bristle patterns characteristic of the two most distal segments of the control. In a few such cases a partial intersegmental membrane was present in the regenerated segment. Removal of the tarsus resulted in a structurally complete regenerate which was smaller than the control tarsus. The largest leg regenerates were obtained when amputation was performed through the tibia. With amputation through the femur, a decrease in length of the remainder of this segment was observed after the first ecdysis. This type of amputation and amputation through the trochanter in some cases resulted in the formation of a globular stump containing tarsal claws. The results indicate that amputation of part of an appendage in Oncopeltus does not stimulate an increased growth rate in the stump, but merely causes reorganization of the stump material which subsequently grows at the normal rate. Since even the most hypomorphic regenerates contained well-formed claws, even though proximal parts were missing, it appears that the reorganization process must begin at the most distal point and proceed proximally.     

Supernumerary regeneration in the large milkweed bug Oncopeltus fasciatus.

As an approach to the problem of pattern formation in the insect appendage, various graft combinations were studied in the legs of the large milkweed bug Oncopeltus fasciatus. Metathoracic legs of fourth instar larvae were amputated through the tibia within 24 hr after ecdysis and grafted back onto the stumps. The orientation of the graft was altered by rotation through 90 or 180° and/or by exchanging right and left stumps and grafts, yielding seven possible orientations in addition to the control. Many of these grafts resulted in the production of one or two supernumerary regenerates of the distal segments, which appeared at the graft junction after the second postoperative ecdysis. When two supernumerary regenerates resulted, one appeared to be produced from the stump and the other from the graft. When one regenerate was present, it appeared to be a composite of material produced from both the stump and the graft. In contrast to the results obtained in cockroaches, the external face of the leg appeared to be the only one capable of giving rise to a supernumerary regenerate.     

Studies on the pteridines in the milkweed bug,Oncopeltus fasciatus (Dallas)

A quantitative re-examination of the pteridines in Oncopeltus has given results at variance with those published previously. The precursors of the red pteridine, erythropterin, are xanthopterin and, probably, oxalacetic acid. Xanthopterin is not present as such in Oncopeltus eggs but exists as 7,8-dihydroxanthopterin. Chrysopterin and biopterin have also been isolated and identified from later growth stages of the bug.     

Intercalary leg regeneration in the large milkweed bug Oncopeltus fasciatus.

Grafting operations were performed on the metathoracic legs of fourth instar Oncopeltus fasciatus within 24 hr after ecdysis. Different levels along the tibia were combined so that a lengthened tibia, approximately 1.5 times the normal length, or a shortened tibia, about half the normal length, were created. Intercalary regeneration occurred between the graft and stump in both combinations and the extra tissue was visible on the adult leg. The intercalary regenerate produced by the lengthened tibia showed reversed bristle polarity, while that produced in the shortened segment showed normal polarity. It is suggested that a regenerate with reversed polarity represents a mirror image duplication of the graft and might originate from the graft, whereas a regenerate with normal polarity may originate, as in normal regeneration, from the stump. It appears that each level in the appendage has the developmental capacity to produce only more distal structures. This conclusion is supported by the results of a grafting operation in which a portion of the tibia was grafted back on to the stump with its proximo-distal axis reversed. Regeneration of appropriate distal structures proceeded from the free proximal surface of the grafted tibia.     

Utilization of sugars by the large milkweed bug, Oncopeltus fasciatus (Hemiptera: Lygaeidae) maintained on chemically defined diets

The utilization of carbohydrates by the large milkweed bug, Oncopeltus fasciatus (Dallas), was studied by feeding the nymphs on chemically defined diets and recording the effects of the various sugars on growth, development, survival, and reproduction. The carbohydrate utilization by the early nymphal stages was largely similar to that observed for later ones. The results showed that some dietary carbohydrate is necessary for maintaining O. fasciatus on chemical diets. Arabinose and rhamnose were of poor value, but the insect was able to utilize glucose, fructose, mannose, galactose, sucrose, maltose, trehalose, lactose, turanose, melibiose, raffinose, and melezitose. The degree of utilization for each sugar, however, differed. Glucose promoted best growth and development among monosaccharides, sucrose among disaccharides, and melezitose among trisaccharides. The optimum level of sugar in the diet was around 4%, and a higher or lower concentration (2, 8, 12%) usually reduced growth. Viable eggs were obtained on some of the diets.

---

Résumé La valeur nutritive de plusieurs glucides pour la punaise de l'asclépiade, Oncopeltus fasciatus, a été déterminée en élevant des larves sur des régimes nutritifs de composition chimique connue. Les effets des sucres sur la croissance, le développement, la longévité et la reproduction ont été établis, et ces effets apparaissent semblables chez les très jeunes larves de même que chez les larves plus âgées. Les résultats indiquent que la présence d'un sucre dans le régime nutritif est indispensable pour la survie de l'insecte. Cependant certains sucres, tels que l'arabinose ou le rhamnose n'ont que peu ou pas de valeur nutritive, tandis que d'autres (glucose, fructose, mannose, galactose, sucrose, maltose, tréhalose, lactose, turanose, mélibiose, raffinose et mélézitose) ont une valeur nutritive variant de faible à élevée. Les sucres qui ont donné les meilleurs résultats sont le glucose, le sucrose, et le mélézitose. La concentration glucidique optimale dans le régime se situe à 4% et une concentration moindre (2%) ou plus élevée (8 ou 12%) ralentit généralement la croissance. Les larves sur certains régimes se sont développées jusqu'au stade adulte et des ufs fertiles ont été pondus.

     

Embryology of the thoracic and abdominal ganglia of the large milkweed bug, Oncopeltus fasciatus (Dallas), (Hemiptera, Lygaeidae)

In this study, the condensation of the three thoracic and 11 abdominal segmental ganglia to form a prothoracic and central nerve mass during embryogenesis is described. During katatrepsis, many changes occur in the organization of these ganglia; this study suggests that some of these changes are caused by mechanical forces acting on the ventral nerve cord at this time. The ventral nerve cord begins its anterior migration and coalescence ten hours after katatrepsis and is completed 63 hours later. The central ganglion is made up of the meso- and metathoracic ganglia and seven abdominal ganglia. Intrasegmental median cord nuclei are shown to form glial elements in the median sagittal plane of the neuropile and in the longitudinal connectives. Intersegmental median cord neuroblasts migrate into the posterior gangliomeres but, apparently, degenerate soon after katatrepsis. Lateral cord cells bordering on the neuropile form a glial investment that surrounds this fiber tract region. Peripheral lateral cord cells are shown to form the cells of the outer ganglionic sheath, the perineurium.     

Pteridines in the milkweed bug,Oncopeltus fasciatus (Dallas): I. Identification and localization

The milkweed bug, Oncopeltus fasciatus (Dallas), contains 3 pteridines, xanthopterin, isoxanthopterin, and 2-amino-4-hydroxypteridine. The 2-amino-4-hydroxypteridine is found only in the midgut; isoxanthopterin and bound xanthopterin are present in all body tissues and excrement throughout the life cycle. Xanthopterin may be freed by heat or acid or base treatment. Pteridines are present in members of 4 subfamilies of the Scutelleridae.     

Identification of the adipokinetic hormone of the large milkweed bug Oncopeltus fasciatus

Abstract The adipokinetic hormone (AKH) of the large milkweed bug Oncopeltus fasciatus is isolated from an acidified methanolic extract of 200 corpora cardiaca, purified by single step reversed phase high‐performance liquid chromatography (HPLC) and N‐terminally deblocked using pyroglutamate aminopeptidase. The sequence is identified by Edman degradation and matrix assisted laser desorption/ionization‐time of flight mass spectroscopy as pGlu‐Leu‐Asn‐Phe‐Ser‐Pro‐Asn‐Trp amide. This structure is confirmed by chemical synthesis and coelution of native and synthetic peptide on HPLC. The AKH of O. fasciatus is identical to Tenmo‐HrTH, a member of the adipokinetic/red pigment‐concentrating hormone peptide family that had been isolated earlier from several tenebrionid beetles. Tenmo‐HrTH causes a significant rise in the concentration of haemolymph lipids when injected into adult male and female O. fasciatus, but displays no hyperglycaemic activity. There is no indication of the presence of other AKHs in O. fasciatus. The large milkweed bug represents the first member of the seed bugs (Lygaeidae) for which the endogenous AKH has been identified.     

Ouabain-resistant Na,K-ATPases and cardenolide tolerance in the large milkweed bug,Oncopeltus fasciatus

Oncopeltus fasciatus tolerated 1954× and 7288×, respectively, the LD₅₀ ouabain dose of Schistocerca gregaria and Periplaneta americana when ouabain was injected into the haemocoel of these insects. The maximal ouabain dose that could be injected into O. fasciatus (200 nmol) resulted in no mortality; this dose is higher than the lethal ouabain doses recorded for vertebrates and invertebrates. The ouabain concentration resulting in 50% inhibition (I₅₀) of Na,K-ATPase activity was determined in lyophilates of nervous tissue of O. fasciatus and brain and recta of S. gregaria and were 2.0 × 10⁻⁴, 2.0 × 10⁻⁶ and 1.0 × 10⁻⁶ M, respectively. The I₅₀ value for ouabain inhibition of Na,K-ATPase activity in the nervous tissue of O. fasciatus is higher than the I₅₀ values for nervous tissue in most other insects as well as many other invertebrate and vertebrate tissues. Thus, the presence of ouabain-resistant Na,K-ATPases appears to be a factor in the tolerance and sequestration of plant cardenolides in O. fasciatus.     

Synthesis of 5 S and transfer RNA during embryogenesis in the milkweed bug, Oncopeltus fasciatus

Synthesis of 5 S RNA and transfer RNA has been shown to begin at the onset of gastrulation in eggs of the milkweed bug, Oncopeltus fasciatus. 5 S RNA synthesis parallels that of ribosomal RNA, but transfer RNA synthesis is somewhat different.     

Feeding, nutrient flow, and digestive enzyme release in the giant milkweed bug, Oncopeltus fasciatus

Abstract.  The giant milkweed bug, Oncopeltus fasciatus , is an intermittent feeder, spending most of its time in compact groups away from the food, with feeding bouts lasting 30–60 min. They macerate a small area of the seed with their stylets, secreting large amounts of saliva (which suspends and emulsifies the seed tissue), and quickly suck the emulsion into the gut. Therefore, little digestive activity occurs in the seed tissue. Freshly ingested food in the first gut region (V1) consists of 50% water, 46% lipid, 5% protein, and 1% soluble carbohydrates. The junction of the midgut and hindgut lacks a lumen in the larvae stages, and a 3 μL large oil drop (> 85% triacylglycerides) accumulates in the third gut region (V3). Approximately 24 h after the imaginal moult, this large volume of oil and faeces is defecated, and thereafter an oily faeces is defecated several times daily. It is postulated that the delayed egestion in larvae protects them against suffocation due to self-soiling with oily faeces. Serine proteases are absent in the gut and, instead, a cystein proteinase, inhibited by E64, is present (gut pH 6.0). High lipase activity was found throughout the gut, but only limited amylase activities. Basal levels of all three enzymes occurred throughout the gut, but at lower levels in starved bugs. Salivary gland extracts contain a cysteine proteinase and a lipase, but no amylase.