The role of heliothine hairpencil compounds in female Heliothis virescens (Lepidoptera: Noctuidae) behavior and mate acceptance

Studies on numerous insect species suggest that male-produced sex pheromones play a role in attracting females; as aphrodisiacs, making females more quiescent; or as a means of inhibiting competing males. Male heliothine moths display abdominal hairpencils during courtship, but the specific effects of the odors released on female behavior have not yet been elucidated. This study investigates the role of male hairpencil compounds in female Heliothis virescens mating behavior. Female H. virescens were exposed to filter paper loaded with hairpencil extracts of male H. virescens, Heliothis subflexa and Helicoverpa zea, and observed for behavioral responses to odors. Single synthetic compounds found in the H. virescens hairpencil blend were also tested. In mating assays between single male and female H. virescens it was found that: (i) antennectomized females mated less frequently than sham-operated females; (ii) females mated less frequently with males whose hairpencils had been surgically removed; (iii) females mated with males with ablated hairpencils if a filter paper loaded with one male equivalent of H. virescens hairpencil extract was presented simultaneously; and (iv) this effect was species-specific, as presentation of H. subflexa or H. zea hairpencil extracts did not restore mate acceptance. This study suggests that odors released by male hairpencils are important in mate acceptance by female H. virescens, and may play a role in mate choice and species isolation.     

Behaviorally mediated contacts between scent organs: Another prerequisite for pheromone production inDanaus chrysippus males (lepidoptera)

Summary Males ofDanaus butterflies possess two binate glandular organs: abdominal hairpencils, which release aphrodisiac pheromones during courtship behavior, and pockets on the hindwings. Between these two types of organs contacts are established behaviorally: the hairpencils are dipped into the alar pockets (Fig. 2). GLC analyses of hairpencil and wing pocket extracts fromDanaus chrysippus show that this contact behavior is a prerequisite for the synthesis of the ketonic pheromone component (for which the term danaidone is suggested here) in physiologically normal amounts. Danaidone occurs on the hairpencils only i) after the males have ingested pyrrolizidine alkaloids as precursors, and ii) after the hairpencils have been dipped into the wing pockets. The appearance of danaidone in the wing pockets also depends on the ingestion of alkaloidal precursor, but is not affected by the above mentioned contact behavior. Mechanisms by which contact behavior might control pheromone biosynthesis are discussed.We appreciate the skillful and reliable greenhouse and laboratory assistance of C. Adrian and H. Mayr, and we thank Drs. R.A. Steinbrecht and P. Lounibos for kindly supplying us with danaids from Kenya. Partial support of this research by Grant No. AI-12020, from the National Institutes of Health (USA), is acknowledged with pleasure.     

Differentiation in the cocoonase producing silkmoth galea: ultrastructural studies.

The differentiation of the cocoonase zymogen-producing cell groups (organules) is described at the ultrastructural level. Two types of zymogen cells arise early in development. One cell of each type associates specifically with the other and with a single duct cell to form an organule. Each organule elaborates a complex cuticular duct apparatus, in a morphogenetic process that involves specific attachments and movements of the apical cell regions, formation of a pseudoflagellar scaffolding, secretion of cuticular material, and extracellular reorganization of this material. Halfway through development, the zymogen cells retract from the duct apparatus, and th resulting extracellular space (storage vacuole) expands as it fills with cocoonase zymogen. The ultrastructure of the zymogen cells is discussed in terms of their growth and sequential secretory activities. Comparison is made with other insect organules.     

The structure and development of the hairpencil glands in males of the queen butterfly, Danaus gilippus berenice

Queen butterflies do not mate until the male has brushed the tufts of his scented, abdominal ”?hairpencils? over the female's head and antennae. The trichogen cells located at the base of each hairpencil are secretory. Presumably, these cells produce the sex pheromone necessary for mating. The liquid secretion must move from a central, microvillus-lined vesicle through the cuticle of the hairs to coat numerous, free, cuticular ?dust”? particles which adhere to the hairs' surface. The dust carries the secretion to or near the female's antennae. In the pupal stage the dust particles develop as outpocketings of the hair epicuticle. An amorphous matrix, probably protein epicuticle, is deposited in the outpocketings between the cuticulin layer and plasma membrane of the hair. Before the butterfly emerges from the pupa the matrix becomes enclosed by cuticulin, and the particles pinch off from the hair.     

棉铃虫雄性定向抑制剂的研究:雄性信息素组分化学结构鉴定(英文)

采用毛细柱气相色谱分析、酸性甲酯化反应和GC-MS证实等手段对棉铃虫雄蛾味刷抽提物进行了结构鉴定。从中鉴定了10种化合物。这些化合物是14:0H,14:Ac,14:COOH,Z11-16:OH,16:0H,16:COOH,16:Ac,18:OH,18:Ac和18:COOH。对各种化合物进行定量分析的结果表明,3种饱和醇在味刷中所占比例高达75％以上。味刷内化合物含量与雄蛾日龄有关,羽化10h以内的雄蛾味刷中不含以上鉴定的各种化合物,经过48h,雄蛾味刷中化合物的量达到高峰,在5日龄以后,化合物含量明显下降。     

马尾松毛虫雄蛾触角毛状感受器的细微结构

马尾松毛虫Dendrolimus punctagus(Walker)雄蛾有一对羽毛状触角。在触角鞭节的每对侧枝的内侧(迎风面)着生许多毛状感受器。每个毛状感受器由几丁质表皮毛及位于其下的三个感觉神经原和三个呈同心排列的辅助细胞-鞘原细胞、毛原细胞和膜原细胞构成。几丁质表皮毛上有许多孔。毛腔内充满感受器淋巴液。感觉神经原发出的树状突伸入毛腔,浸浴于感受器淋巴液内。这些结构特征表明它是一种司嗅觉的化学感受器。雄蛾终生不取食,推断它的嗅觉感受器主要用以感受雌蛾释放的性外激素,帮助寻找配偶。     

Structure and development of the secretory cavities of Myrtus communis leaves

The structure and development of Myrtus communis L. secretory cavities has been studied in young and expanded leaves, using light and scanning electron microscope. Secretory cavities are continuously formed during leaf development, but in mature leaves the rhythm of their appearance shows steep decrease. Each secretory cavity is developed from a single epidermal cell, which undergoes a periclinal division followed by anticlinal and several oblique cell divisions. The lumen of the secretory cavity is initiated by cell wall separation, i.e., schizogenously. The secretory cells line the cavity, where the secreted material is collected. Secretory cavities are covered by modified epidermal cells, which do not seem to form any special aperture. Essential oils seem to be discharged after mechanical treatment of the leaf.     

Persistent fusion pores but transient fusion in alveolar type II cells

The release of vesicle contents following exocytotic fusion is limited by various factors including the size of the fusion pore. Fusion pores are channel-like, narrow structures after formation and proceed through semi-stable states ('fusion pore flickering'), unless they fully expand (full fusion) or close again (transient fusion). Partial release of vesicle contents may occur during transient fusion, which was described to last between milliseconds and seconds, depending on the size of the vesicle. We studied fusion pores in a slow-secreting lung epithelial cell (type II cell) using fluorescence staining of vesicle contents (surfactant) and fluorescence recovery after photobleaching (FRAP). Surfactant is a lipidic material, which is secreted into the alveolar lumen to reduce the surface tension in the lung. We found release of surfactant to be a slow process, which can last for hours. Accordingly, fusion pores in these cells are stable structures, which appear to be a barrier for release. FRAP measurements suggest that transient fusions occasionally take place in these long-lasting fusion pores, resulting in partial release of surfactant into the extracellular space. These data suggest that postfusion mechanisms may regulate the amount of secreted surfactant.     

The Design of Silk Fiber Composition in Moths Has Been Conserved for More Than 150 Million Years

The silk of caterpillars is secreted in the labial glands, stored as a gel in their lumen, and converted into a solid filament during spinning. Heavy chain fibroin (H-fibroin), light chain fibroin (L-fibroin), and P25 protein constitute the filament core in a few species that have been analyzed. Identification of these proteins in Yponomeuta evonymella, a moth from a family which diverged from the rest of Lepidoptera about 150 million years ago, reveals that the mode of filament construction is highly conserved. It is proposed that association of the three proteins is suited for long storage of hydrated silk dope and its rapid conversion to filament. Interactions underlying these processes depend on conserved spacing of critical amino acid residues that are dispersed through the L-fibroin and P25 and assembled in the short ends of the H-fibroin molecule. Strength, elasticity, and other physical properties of the filament are determined by simple amino acid motifs arranged in repetitive modules that build up most of the H-fibroin. H-Fibroin synergy with L-fibroin and P25 does not interfere with motif diversification by which the filament acquires new properties. Several types of motifs in complex repeats occur in the silks used for larval cobwebs and pupal cocoons. Restriction of silk use to cocoon construction in some lepidopteran families has been accompanied by simplification of H-fibroin repeats. An extreme deviation of the silk structure occurs in the Saturniidae silkmoths, which possess modified H-fibroin and lack L-fibroin and P25. [Reviewing Editor: Dr. David Pollok]     

MALE ORIENTATION INHIBITOR OF COTTON BOLLWORM: IDENTIFICATION OF COMPOUNDS PRODUCED BY MALE HAIRPENCIL GLANDS1)

Abstract  Extracts of hairpencils ffom male cotton bollworm moth were analyzed by capillary gas chromatography, acid methanolysis, and GC-MS. Ten components have been identified as_: 14; OH, 14: Ac, 14: COOH, 211–16: OH, 16: OH, 16: Ac, 16: COOH, 18: OH, 18: Ac, and 18: COOH. Based on the statistics of titer of each chemical, the total amount of three saturated alcohols is over 75%. The amounts of the chemicals in the hairpencils are related to the age of males. There are no chemicals identified in the extracts from males less than 10 h after eclosion, then the quantity of compounds increased rapidly during 48 h after adult eclosion. After 5 days, the quantity decreased.     

Cholesterol-dependent cytolysins: from water-soluble state to membrane pore

The cholesterol-dependent cytolysins (CDCs) are a family of bacterial toxins that are important virulence factors for a number of pathogenic Gram-positive bacterial species. CDCs are secreted as soluble, stable monomeric proteins that bind specifically to cholesterol-rich cell membranes, where they assemble into well-defined ring-shaped complexes of around 40 monomers. The complex then undergoes a concerted structural change, driving a large pore through the membrane, potentially lysing the target cell. Understanding the details of this process as the protein transitions from a discrete monomer to a complex, membrane-spanning protein machine is an ongoing challenge. While many of the details have been revealed, there are still questions that remain unanswered. In this review, we present an overview of some of the key features of the structure and function of the CDCs, including the structure of the secreted monomers, the process of interaction with target membranes, and the transition from bound monomers to complete pores. Future directions in CDC research and the potential of CDCs as research tools will also be discussed.     

Biosynthesis and antimicrobial evaluation of backbone-cyclized α-defensins

Defensins are antimicrobial peptides that are important in the innate immune defense of mammals. Upon stimulation by bacterial antigens, enteric α-defensins are secreted into the intestinal lumen where they have potent microbicidal activities. Cryptdin-4 (Crp4) is an α-defensin expressed in Paneth cells of the mouse small intestine and the most bactericidal of the known cryptdin isoforms. The structure of Crp4 consists of a triple-stranded antiparallel β-sheet but lacks three amino acids between the fourth and fifth cysteine residues, making them distinct from other α-defensins. The structure also reveals that the α-amino and C-terminal carboxylic groups are in the proximity of each other (d ≈ 3 ?) in the folded structure. We present here the biosynthesis of backbone-cyclized Crp4 using a modified protein splicing unit or intein. Our data show that cyclized Crp4 can be biosynthesized by using this approach both in vitro and in vivo, although the expression yield was significantly lower when the protein was produced inside the cell. The resulting cyclic defensins retained the native α-defensin fold and showed equivalent or better microbicidal activities against several Gram-positive and Gram-negative bacteria when compared to native Crp4. No detectable hemolytic activity against human red blood cells was observed for either native Crp4 or its cyclized variants. Moreover, both forms of Crp4 also showed high stability to degradation when incubated with human serum. Altogether, these results indicate the potential for backbone-cyclized defensins in the development of novel peptide-based antimicrobial compounds.     

Paneth cell cryptdins act in vitro as apical paracrine regulators of the innate inflammatory response

Intestinal-specific antimicrobial alpha-defensins, termed cryptdins, are secreted into the intestinal lumen by mouse Paneth cells in response to microbial pathogens. Cryptdins kill microbes by forming pores in their limiting membranes. The cryptdin isoforms 2 and 3 also can form anion-conductive pores in eukaryotic cell membranes, thus affecting cell physiology. Here, we find that when applied to apical membranes of the human intestinal cell line T84, cryptdin 3 (Cr3) induces secretion of the proinflammatory cytokine interleukin 8 (IL-8) in a dose-dependent manner. The induction of IL-8 secretion is specific to the cryptdins that form channels in mammalian cell membranes because cryptdin 4, which does not form pores in T84 cells, does not induce IL-8 secretion. Cr3 induces inflammatory cytokine secretion by activating NF-kappaB and p38 mitogen-activated protein kinase in a Ca2+-dependent manner, but influx by extra-cellular Ca2+ is not involved. Unlike other known inflammatory agonists, signal transduction by Cr3 occurs slowly, suggesting a novel mechanism of action. These results show that selective cryptdins may amplify their roles in innate immunity by acting as novel paracrine agonists to coordinate an inflammatory response with the antimicrobial secretions of Paneth cells.     

Characteristics of Structure and Differentiation in the Sieve Element of Lower Vascular Plants

The structure and differentiation of the sieve element of lower vascular plants is reviewed using data obtained primarily from ultrastructural investigations conducted during the last ten years. During the last decade the phloem of representatives from every major group of the ferns and fern allies has been examined with the electron microscope and from these studies a rather clear picture has emerged of the structure of the sieve element protoplast in this diverse group of plants. Present data indicate that although the details of sieve-element differentiation may differ, the protoplasts of the mature sieve elements in the various groups of lower vascular plants are remarkably similar in structure. Each consists of a plasmalemma, a parietal, anastomosing network of smooth ER, plastids, mitochondria and, with the exception of the lycopods, variable numbers of refractive spherules. The protoplasts of mature sieve elements are joined by plasmalemma-lined connections, each arising from a single plasmodesma during the course of sieve element differentiation. The size of the connections in the mature elements range from plasmodesmata-like structures to relatively wide sieve-area pores, depending on the species. Moreover, the contents of the cytoplasmic connections vary somewhat according to the species. Whereas in the lycopods, the sieve-area pores are virtually unoccluded by any cytoplasmic material, the cytoplasmic connections of all other lower vascular plants examined with the electron microscope contain variable amounts of membranous material, apparently tubular elements of ER. In Equi-setum hyemale, Psilotum nudum and the eusporangiate and protoleptosporangiate ferns, the ER membranes are very numerous and virtually occlude the pores. Furthermore, the membranes apparently are not connected with the parietal ER in the lumen of the cell. The sieve-area pores of the leptosporangiate ferns also contain ER membranes, however, they are not as abundant as the membranes of the eusporangiate and protoleptosporangiate ferns. In addition, in the leptosporangiate ferns the pore membranes apparently are united with the parietal ER in the lumen of the cell.     

The netrin protein family

The name netrin is derived from the Sanskrit Netr, meaning ''guide''. Netrins are a family of extracellular proteins that direct cell and axon migration during embryogenesis. Three secreted netrins (netrins 1, 3 and 4), and two glycosylphosphatidylinositol (GPI)-anchored membrane proteins, netrins G1 and G2, have been identified in mammals. The secreted netrins are bifunctional, acting as attractants for some cell types and repellents for others. Receptors for the secreted netrins include the Deleted in Colorectal Cancer (DCC) family, the Down''s syndrome cell adhesion molecule (DSCAM), and the UNC-5 homolog family: Unc5A, B, C and D in mammals. Netrin Gs do not appear to interact with these receptors, but regulate synaptic interactions between neurons by binding to the transmembrane netrin G ligands NGL1 and 2. The chemotropic function of secreted netrins has been best characterized with regard to axon guidance during the development of the nervous system. Extending axons are tipped by a flattened, membranous structure called the growth cone. Multiple extracellular guidance cues direct axonal growth cones to their ultimate targets where synapses form. Such cues can be locally derived (short-range), or can be secreted diffusible cues that allow target cells to signal axons from a distance (long-range). The secreted netrins function as short-range and long-range guidance cues in different circumstances. In addition to directing cell migration, functional roles for netrins have been identified in the regulation of cell adhesion, the maturation of cell morphology, cell survival and tumorigenesis.     

Enteric defensins: antibiotic peptide components of intestinal host defense

Five intestinal defensins, termed cryptdins 1-5, have been purified from mouse small bowel, sequenced, and localized to the epithelium by immunohistochemistry. Although identified as members of the defensin peptide family by peptide sequencing, enteric defensins are novel in that four cryptdins have amino termini which are three to six residues longer than those of leukocyte-derived defensins. A fifth cryptdin is the first defensin to diverge from the previously invariant spacing of cysteines in the peptide structure. The most abundant enteric defensin, cryptdin-1, had antimicrobial activity against an attenuated phoP mutant of Salmonella typhimurium but was not active against the virulent wild-type parent. Immunohistochemical localization demonstrated that cryptdin-1, and probably cryptdins 2 and 3, occur exclusively in Paneth cells, where the peptides appear to be associated with cytoplasmic granules. Biochemical and immunologic analysis of the luminal contents of the small intestine suggest that cryptdin peptides are secreted into the lumen, similar to Paneth cell secretion of lysozyme. The presence of several enteric defensins in the intestinal epithelium, evidence of their presence in the lumen, and the antibacterial activity of cryptdin-1 suggest that these peptides contribute to the antimicrobial barrier function of the small bowel mucosa.     

Cellular mechanisms of netrin function: long-range and short-range actions.

Netrins are secreted proteins that direct axon extension and cell migration during neural development. They are bifunctional cues that act as an attractant for some cell types and as a repellent for others. Several lines of evidence suggest that two classes of receptors, the deleted in colorectal cancer (DCC) family and the UNC-5 family, mediate the attractant and repellent response to netrin. Although netrins were first identified as diffusible long-range cues for developing axons, recent findings provide evidence that they also function as short-range cues close to the surface of the cells that produce them. This short-range function of netrin contributes to guiding neurite outgrowth and mediating cell-cell interactions during development and perhaps also in adults.     

Ricerche Sulle Infrastrutture Cellulari Dello Spadice Di Arum

Abstract

Electronmicroscope studies on fine structure of the ARUM spadix. — Three main stages of spadix development have been recognized. In the first stage — during wigh the growth would be embrional and by cell division, while respiration would sharply increase — new mitochondria are continously generated, mainly by the activity of long organules (chondriochontha). The morphology of these mitochondria is quite peculiar, showing bended cristae, often ring — or arc-shaped, and closed. In the second stage — during wich the growth would be by cell enlargment and respiration would further increase — the production of new mitochondria is decreasing, but thed are still numerous in each cell, and show a larger number of cristae (or tubules) than mitochondria observed in the first stage. A generalized vesicle production by the endoplasmic reticulum and the cristae of mitochondria is apparent at the beginning of the third stage — during wich the spadix gradually involves and respiration would fall down. Later, presumably in concurrence with the loss of cell vitality, the membranes of mitochopdria are disrupted, and all the cell structuree less evident. A great interest is presented by the vesicle production; indeed visicle production by the endoplasmic reticulum, and often also by mitochondria, can be observed in the senescent cells, already present and scattered in the tissues of the very young spadices. This ultimate process is howerer often preceded by the re-absorption of the outer membranes of these organules, in connection of the cristae, wich therefore become opened and in communication with the cell cavity.     

Dynamics of annulate lamellae in Drosophila syncytial embryos

The mitotic events in eukaryotic cells are controlled by a family of evolutionary conserved cyclin-dependent kinases (cdk) that phosphorylate cell proteins, which results in the structural reorganization of the entire cell. Our recent studies of Drosophila syncytial embryos have demonstrated that changes in cdk1 activity controlling the assembly and disassembly of nuclear pore complexes also affect the structure of cytoplasmic pores in annulate lamellae. Here, we report a comparative electron microscopic analysis of the dynamics of these organelles during mitosis throughout the development of a Drosophila syncytial embryo. We presume that the distribution of annulate lamellae containing mature cytoplasmic pores across the cytoplasm reflects local reductions in the mitotic kinase cdk1 activity during the development of Drosophila syncytial embryos.