PROTEIN AND NUCLEIC ACID METABOLISM IN INSECT FAT BODY

1. The appearance of larval fat body as seen under the light or electron microscope depends on the nutritional state of the larva and on the stage of larval development at which the fat body is observed. 2. Early in the last larval instar the cells usually possess a well-developed endo-plasmic reticulum rich in ribosomes, numerous mitochondria, glycogen granules, a Golgi complex and fat droplets, while later in the instar the endoplasmic reticulum is much reduced and mitochondria are few, but glycogen and fat droplets are present in greater amount together with the appearance of large numbers of proteinaceous spheres. 3. Early in the last instar the fat body synthesizes proteins and exports them into the blood, while later in the instar proteins are sequestered from the blood into the fat body. 4. The rate of protein synthesis by the fat body is high in the early to mid part of the last instar, but then falls off rapidly to a low level, at which it remains until the larva pupates. In diapausing pupae, protein synthesis remains at this low level. 5. The similarity between the electrophoretic patterns of proteins from the fat body and those from the blood provides strong evidence that the fat body is the site of synthesis of many of the blood proteins. 6. Some of the blood proteins have been shown to possess enzymic properties, while others are thought to play a role in the transportation of various types of compounds. 7. Ecdysone and juvenile hormone both stimulate the rate of protein synthesis by larval fat body. Protein synthesis in fat body from diapausing pupae is stimulated after injury to the pupae. 8. The appearance of adult fat body and the amount of protein it contains is often closely linked with the nutritional and reproductive states of the insect. 9. An important role of the fat body in the adult female insect is the synthesis of yolk proteins, which are released into the blood and then taken up by the developing oocytes. This synthesis and uptake are under the control of hormones secreted by the corpora allata and by the median neurosecretory cells of the pars intercerebralis. 10. The RNA content of fat body in final-instar larvae is not constant throughout the instar. In some larvae it is at its highest level early in the instar, falling to a low level as the instar progresses, while in other larvae (e.g. Calliphora) the level of RNA in fat body does not decrease as the instar progresses. 11. In some dipterous insects the base composition of total RNA is DNA-like in that the guanine + cytosine content is low, accounting for 40 % of the bases. A similar composition is seen in rapidly labelled RNA isolated from insects of other orders (Coleoptera and Lepidoptera), but the base content of total RNA from these latter insects resembles ribosomal RNA from vertebrate tissues in that it has a high (ca. 60 %) guanine + cytosine content. 12. The RNA/DNA ratios in blowfly larval tissues are high compared with those found in any vertebrate tissue. 13. In larval fat body, RNA synthesis is low at the time of a moult, increases during the early and mid-instar period and subsequently falls during the latter part of the instar. During the pupal period, especially during pupal diapause, the rate of RNA synthesis is very low and then increases during the subsequent development of the pharate adult. Injury to diapausing pupae results in an increased rate of RNA synthesis in most of their tissues. 14. Ecdysone and juvenile hormone both stimulate RNA and DNA synthesis in larval and adult fat body and in other tissues, although there is evidence that in some tissues these two hormones may act antagonistically to each other. The insecticide DDT also has been shown to stimulate RNA synthesis in tissues of adult insects.     

拟南芥菜(Arabidopsis thaliana)子叶中蛋白体形成的超微结构和免疫电镜定位研究

本文对拟南芥菜(Arabidopsis thaliana)种子发育过程中贮藏蛋白的积累和蛋白体的形成进行了超微结构和免疫电镜定位的研究。常规超薄切片的电镜观察表明,在开花后第10天(10 DAF),高电子密度的蛋白质物质开始在子叶细胞的液泡中沉积。这一过程一直延续到种子接近成熟(14 DAF),这时液泡中充满了蛋白质物质,转变成为大的蛋白体。利用了该种植物主要种子贮藏蛋白之一的12 s球蛋白的单克隆抗体作为免疫探针,以蛋白质A-胶体金电镜技术对12 s种子蛋白进行了细胞内定位,证实了在液泡中积累的物质为种子贮藏蛋白。实验结果表明在拟南芥菜中,子叶细胞中的液泡是蛋白体的前体,肯定了蛋白体的发生起源于液泡的观点。本文还对应用胶体金电镜技术进行细胞内定位的某些问题作了初步探讨。     

THE STRUCTURE OF EOSINOPHIL LEUKOCYTE GRANULES IN RODENTS AND IN MAN

The structure of the specific granules of eosinophil leukocytes has been studied by electron microscopy in sections of tissues, buffy coats, and sediments of peritoneal washings of rats, mice, guinea pigs, and men. The core of eosinophil granules is a crystal which has a cubic lattice with a repeat of ~30 A in rodents and ~40 A in man. The chemical composition of the core is discussed in connection with recent cell fractionation studies, and the hypothesis that the core is a crystal of peroxidase is considered.     

THE STRUCTURE OF INTERSEGMENTAL MUSCLE FIBERS IN AN INSECT, PERIPLANETA AMERICANA L

The organization of intersegmental muscle fibers associated with the dorsal abdominal sclerites of the cockroach is described. These fibers correspond closely, in the disposition and derivation of the membranes of the transverse tubular system and sarcoplasmic reticulum cisternae, with insect synchronous flight muscle fibers, but differ markedly from these in their fibrillar architecture and mitochondrial content. The mitochondria are small and generally aligned alongside the prominent I bands of the sarcomere, and, in the best-oriented profiles of the A bands, thick filaments are associated with orbitals of twelve thin filaments, a configuration that has also been observed in striated fibers of insect visceral muscle. These structural features of insect muscles are compared and discussed in terms of possible variations in the control of contraction and relaxation, and in the nature of their mechanical role.     

ORIGIN AND DEVELOPMENT OF PROTEIN GRANULES IN MAIZE ENDOSPERM

Protein granule development in the starchy endosperm of normal maize (inbred line, W46A) was studied with optical and electron microscopy. The granules were first observed 12 days after pollination as spherical deposits, usually single, within an enclosing membrane. They developed from vesicles produced by the endoplasmic reticulum and were formed both as small localized cisternal dilations and as enlargements at the ends of endoplasmic reticulum. Dictyosomes also appear to proliferate protein granule vesicles. The protein accumulated in the granules appears to be synthesized outside the membranes. Once initiated, the granules rapidly increase in size and number as the kernel develops; their final diameter ranges up to 2 μ. Immature granules stain with a variety of biological dyes in aqueous solution as well as with metal stains; mature granules stain lightly or not at all. At kernel maturity, the protein granules are embedded in a protein matrix.     

PROTEIN SYNTHESIS IN THE FAT BODY OF THE FIFTH LARVAL INSTAR PHILOSAMIA CYNTHIA LARVAE-RELATION TO TWO FEEDING PERIODS

In order to investigate the determinism of the metabolic variations associated with developmental change, protein synthesis was measured in vivo , in the fat body of Philosamia cynthia. During the fifth larval instar two periods, the necessary feeding period and the facultative feeding period, which constitute decisive developmental periods of the insect were xamined. eAfter incorporation of [³H]-glycine protein synthesis was estimated by a calculation which takes account of the dilution of the radioactive amino acids in the free amino acid pool. During the necessary feeding period (from ecdysis to day 4) the protein synthesis increased steadily whereas during the facultative feeding period (from day 4 to onset of spinning) the protein synthetic level decreased. Some preliminary hypotheses are suggested for the mechanism responsible for protein synthesis variations during a larval instar.     

CELL STRUCTURE AND THE METABOLISM OF INSECT FLIGHT MUSCLE

The biochemical properties of insect flight muscle were investigated to ascertain the mechanisms whereby energy is made available for the contractile processes. It was found: 1. The endogenous respiration of muscle homogenates was diminished by starving the flies. The substrate for this respiration was probably glycogen. 2. To obtain the maximal rate of oxidation of glucose, the homogenate had to be fortified with inorganic phosphate, Mg ions, ATP, and cytochrome c. The nucleotides, AMP and ADP, were not as effective as ATP. The addition of DPN or TPN was not necessary for this system. 3. Flight muscle homogenates oxidized glycogen, some sugars, and amino acids, as well as the intermediates of the glycolytic and tricarboxylic acid cycles. Other evidence demonstrated the substrate specificity of the muscle. 4. By centrifugation, the muscle homogenate was divided into two fractions: one, a soluble fraction representing the sarcoplasm; the other, the particulate fraction which contained the fibrils and the sarcosomes. 5. The particulate fraction, alone, oxidized all the citric acid cycle intermediates, α-glycerophosphate, phosphopyruvate, and the amino acids, glutamic, proline, and cysteine. Regardless of the substrate, no oxygen uptake was found with the sarcoplasm by itself. 6. A recombination of the sarcoplasm and the particulate component was required for the oxidation of glycogen, the hexoses, and all the phosphorylated intermediates of glycolysis, except phosphopyruvate. 7. Isolated mitochondria accounted for all the enzymatic activity of the particulate fraction. These results demonstrate that the enzymes of intermediate metabolism are localized in the sarcoplasm or sarcosomes. The third cytological entity, the myofibrils, plays no role in the energy-providing scheme. From a functional viewpoint, the sarcoplasm and the mitochondria, in combination, furnish the energy for the actomyosin contraction. The results are discussed in relation to analogous findings in other insects and vertebrates.     

AN AUTORADIOGRAPHIC AND ULTRASTRUCTURAL STUDY OF GLYCOGEN METABOLISM AND FUNCTION IN THE ADULT FAT BODY OF A BLACK-FLY DURING OÖGENESIS

Glycogen was abundant in fat-body cells of 1-day-old autogenous black-flies (Simulium vittatum Zett.) as seen by electron-microscopy, and decreased during the next 2 days. However, the ability of the fat body to synthesize glycogen remained undiminished. More C¹⁴-glucose was incorporated in vivo with added carrier glucose than without. The percentage of diastase-resistant particles, as seen by autoradiography, was higher when C¹⁴-glucose was injected without the carrier. Along with the abundance of diastase-resistant radioactive particles, the dense material enclosed in the Golgi vesicles, plus the well-developed endoplasmic reticulum and numerous Golgi complexes, suggested that glycogen in the fat-body cell provides carbohydrates for glycoprotein synthesis and export, as well as for an endogenous energy source.

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Zusammenfassung Wie im Elektronenmikroskop zu sehen ist, führen einen Tag alte Weibchen der autogenen, nordamerikanischen Kriebelmücke, Simulium vittatum Zett., in den Zellen des Fettkörpers reichlich Glykogen. Obwohl der Glykogengehalt während der nächsten zwei Tage der Oogenese rasch abnimmt, behält der Fettkörper unvermindert die Fähigkeit zur Glykogensynthese, wie der rasche Einbau von ¹⁴C-Glukose in vivo beweist. Bei Zugabe von Trägerglukose wurde mehr ¹⁴C-Glukose als ohne diese eingebaut. Jedoch war der Prozentsatz diastaseresistenter Partikel höher, wenn ¹⁴C-Glukose ohne Carrier injiziert wurde, wie autoradiographisch zu erkennen war.Die Häufigkeit dieser diastaseresistenten Partikel und das in den Golgi-Bläschen eingeschlossene elektronendichte Material sowie die zahlreichen Golgi-Komplexe in enger Nachbarschaft der Glykogen-Region und das wohlentwickelte endoplasmatische Retikulum lassen vermuten, daß Glykogen in den Fettkörperzellen das Kohlenwasserstoff-Reservoir für die Glykoprotein-Synthese und-Lieferung darstellt. Die ausschließliche Verbreitung von Mitochondrien in der Glykogen-Region, welche ebenfalls eng mit dem endoplasmatischen Retikulum verbunden ist, dürfte darauf hinweisen, daß Glykogen außerdem die endogene Energiequelle für den Stoffwechsel in den Fettkörperzellen darstellt.

     

THE EFFECTS OF RESERPINE AND HYPOXIA ON THE AMINE-STORING GRANULES OF THE HAMSTER CAROTID BODY

The carotid bodies from control, reserpine-treated, and hypoxia-treated hamsters were fixed with phosphate-buffered glutaraldehyde and osmium tetroxide, s-Collidine-buffered osmium tetroxide, or phosphate-buffered glutaraldehyde followed by potassium dichromate incubation. Following glutaraldehyde-osmium tetroxide fixation no differences in density or population of the electron-opaque granules in the glomus cells of either control or experimental animals were observed. With s-Collidine-buffered osmium tetroxide and the glutaraldehyde-dichromate technique a marked decrease in density without an appreciable reduction in number of granules was noted after reserpine treatment, while in hypoxia-treated hamsters the density and population of the granules were not different from those of the controls. The results indicate that reserpine depletes the amines without granule disappearance and that hypoxia does not affect the amine content of the granules. It is suggested that following glutaraldehyde-osmium tetroxide double fixation, persistence of the density of the granules in reserpine-treated animals is due primarily to the nonamine content, and that the amines in the glomus cells are probably not directly involved in the respiratory reflex.     

THE STRUCTURE OF INSECT VIRUS PARTICLES

Thin sections have been cut of the virus particles from four types of insect virus diseases: cytoplasmic polyhedroses of lepidopterous larvae, a nuclear polyhedrosis of Tipula paludosa (Diptera), a granulosis from Melanchra persicariae (Lepidoptera), and a new virus disease without polyhedra from T. paludosa. The cytoplasmic polyhedral viruses are thought to have composite particles in some cases. The shape and enveloping membranes of the different virus particles are compared. In the new virus disease of T. paludosa some of the virus particles appear to be empty; inclusion bodies surrounded by complicated membranes are also demonstrated.     

ON THE MOLECULAR STRUCTURE OF STARCH GRANULES

It is a problem how to fit the molecules of amylose and amylopectin into the ultrastructural apposition layers of starch granules which have a width of only 0.1μ. Stretched amylose chains with a length up to 1 μ can be ruled out. In addition to the proposal of Miihlethaler, who visualizes folded amylopectin chains, a model with a helical amylose chain is discussed. A helix with six glucose residues per turn would be compatible with the crystal lattice found by Kreger; and the disorder caused by this helix in the hexagonal lattice of the parallel side chains of the amylopectin would account for the low optical anisotropy of the starch granules as compared to the parallel β-polyglucosan chains in cellulose. The proposed model, fitted into the 0.1 μ apposition ring of the starch granule, shows tangential strata with a layer repeat of about 80 A and a radial chain lattice with the fibre period 10.6 A; i.e., it shows a combination of a lamellar and a fibrous structure.     

WALL STRUCTURE AND LATERAL FORMATION IN THE ALGA BRYOPSIS

Green , Paul B. (U. Pennsylvania, Philadelphia.) Wall structure and lateral formation in the alga Bryopsis. Amer. Jour. Bot. 47(6) : 476–481. Illus. 1960.—The large multinucleate cells of Bryopsis pennata regularly produce laterals at the apical tip. Cells were freed of protoplasm and the remaining cell wall was flattened and studied in polarized light and interference microscopes. Both the cylindrical main axis and the cylindrical laterals are negatively birefringent and thus apparently have generally a transverse arrangement of microfibrils. Lateral formation is first seen in the formation of round fields of concentric microfibrils on the main axis which persist as lateral-bases. The center of a field protrudes, making a small out-pouching of the main axis. The thin-walled protrusion increases first mostly in diameter and then in length to make a lateral. There is a definite correlation between wall structure (optical anisotropy) and cell shape in Bryopsis.     

THE STRUCTURE OF THE SMOOTH MUSCLE FIBRES IN THE BODY WALL OF THE EARTHWORM

1. The structure of the smooth muscle fibres in the longitudinal muscle coat of the body wall of Lumbricus terrestris has been investigated by phase contrast light microscopy and electron microscopy. 2. The muscle fibre is ribbon-shaped, and attached to each of its two surfaces is a set of myofibrils. These are also ribbon-shaped, and they lie with their surfaces perpendicular to the surfaces of the fibre, and their inner edges nearly meeting in the middle of the fibre. These fibrils are oriented at an angle to the fibre axis, and diminish greatly in width as they approach the edge of the fibre. The orientation of the set of fibrils belonging to one surface of the fibre is the mirror image of that of the set belonging to the other surface; thus, when both sets are in view in a fibre lying flat on one face, the fibre exhibits double oblique striation. A comparison of extended and contracted fibres indicates that as the fibre contracts, the angle made between fibre and fibril axes increases (e.g. from 5 to 30°) and so does the angle made between the two sets of fibrils (e.g. from 10 to 60°). 3. The myofibril, throughout its length, contains irregularly packed filaments, commonly 250 A in diameter, which are parallel to its long axis and remain straight in contracted muscles. Between them is material which probably consists of much finer filaments. Thus A and I bands are absent. 4. Bound to one face of each fibril, but not penetrating inside it, is a regularly spaced series of transverse stripes. They are of two kinds, alternating along the length of the fibril, and it is suggested that they are comparable to the Z and M lines of a cross-striated fibril. The spacing of these stripes is about 0.5 µ ("Z" to "Z") in extended muscles, and 0.25 µ in contracted muscles. A bridge extends from each stripe across to the stripeless surface of the next fibril.