The accessory gland proteins in maleDrosophila: structural,reproductive, and evolutionary aspects

Recent results from biochemical and molecular genetic studies of the accessory gland proteins in maleDrosophila are reviewed. The most prominent feature is the species-specific variability. However, the analysis of the sex peptide inD. melanogaster shows that there is a strong homology in the molecular structure to the closely related sibling species, and that divergence increases with increasing phylogenetic distance. For this reason the sex peptide, after being transferred to the female genital tract during copulation, reduces receptivity and increases oviposition only in virgin females belonging to the same species group and subgroup. Even though studies were hitherto limited to a small number of the secretory components, it is evident that the accessory gland proteins play a key role in reproductive success of the fruit fly by changing female sexual behavior, supporting sperm transfer, storage and displacement. Thus, genes encoding the accessory gland proteins are apparently under strong evolutionary selection.     

Distribution of (Na++K+)-ATPase in the hindgut ofGlossina morsitans morsitans Westwood

Summary (Na⁺+K⁺)-ATPase activity was higher in preparations from the ileum ofGlossina mortisans than in those from the rectum. This result suggests that the ileum as well as the rectum, may play a role in osmoregulation in the tsetse fly.     

Precocene treatment of the female tsetse flyGlossina morsitans morsitans sterilises her female offspring

Summary When adult femaleGlossina morsitans morsitans were treated with either precocene I or precocene II, females of the F₁ generation, emerging from pupae of the 2nd, 3rd and 4th reproductive cycles failed to develop oocytes.Acknowledgments. The authors wish to thank Dr G. C. Unnithan for helpful discussions.     

Sexual dimorphism in the lipid fraction of the defensive secretion ofGastrophysa viridula (Coleoptera: Chrysomelidae)

The lipid fraction of the defensive secretion of adultGastrophysa viridula was analyzed by gas-liquid chromatography and gas chromatography-mass spectrometry. Three long chain acetates and two methyl-branched hydrocarbons were identified and their amounts in the secretion of single field-collected and laboratory-reared beetles determined. The composition of the secretion was affected by the sex, reproductive status, and age of the beetle. The ratio of (Z)-11-eicosenyl acetate to (Z)-13-docosenyl acetate ranged from 0.30 to 0.78 in male beetles and from 1.05 to 2.06 in female beetles. The biological significance of this sexual dimorphism is discussed.     

Tissue- and stage-specific RNA patterns inDrosophila as revealed by gradient formamide gel electrophoresis

Summary The in vitro labeled RNAs in the accessory glands from male adults, and in the salivary glands and fat body from developing larvae ofDrosophila melanogaster were separated on linear gradient acrylamide gels in formamide. The patterns appear to be tissue- and stage-specific, and several lines of evidence indicate that the visualized radioactive bands include both hnRNA and messages.Acknowledgments. The financial support by the Swiss National Science Foundation and the George and Antoine Claraz-Schenkung is gratefully acknowledged. I thank Prof. Max Birnstiel for providing me with the sea urchin histone mRNA and Prof. Martin Billeter for providing me with the rabbit globin mRNA as RNA markers.     

Cytogenetic studies of Hynobiidae (Urodela). IX. Karyological characteristics ofHynobius abei Sato by means of R-and C-banding

Summary Chromosomal characteristics of the salamander speciesHynobius abei, from Ohimya (Kyoto) were revealed by the techniques of R-and C-banding. The karyotype ofH. abei was characterized by the shortness of an R-negative (C-positive) band in the terminal region of the long arm of chromosome 2 and a band encompassing the whole short arm of chromosome 10. These two bands inH. abei were the shortest among those of the variousHynobius species that have been examined. Otherwise no differences could be detected betweenH. abei and seven other pond-type species ofHynobius (2n=56) in terms of the banding patterns of 18 specifically identifiable pairs of their chromosomes.     

Spermatogenesis is extraordinarily accelerated in metamorphosis-arrested larvae of a salamander,Hynobius retardatus

Laboratory experiments were conducted to induce neoteny inHynobius retardatus, which had been reported to propagate in larval forms like axolotl. A large number of newly hatched larvae were reared in an aqueous solution of thiourea (TU) and sodium perchlorate (SPC) in order to arrest the metamorphosis. Gonadal development in the metamophosis-arrested larvae was compared with that in normally metamorphosing and metamorphosed controls. Metamorphosis-arrested male larvae produced morphologically mature spermatozoa approximately 4 months after hatching, when the gonads in the controls began to differentiate into testes, or to show the premeiotic proliferation of germ cells. Possible endocrine controls of these phenomena are discussed.     

Tissue- and stage-specific RNA patterns in Drosophila as revealed by gradient formamide gel electrophoresis

The in vitro labeled RNAs in the accessory glands from male adults, and in the salivary glands and fat body from developing larvae of Drosophila melanogaster were separated on linear gradient acrylamide gels in formamide. The patterns appear to be tissue- and stage-specific, and several lines of evidence indicate that the visualized radioactive bands include both hnRNA and messages.     

Evolutionary and functional epitopes of the Spätzle protein: New insights into activation of the Toll receptor

Sp?tzle, a dimeric ligand, binds to the Drosophila Toll receptor and activates the signal pathway functioning in both embryonic patterning and innate immunity. Here, we used the evolutionary trace approach based on phylogenetic information to predict the evolutionary epitope of Sp?tzle and found that it mainly clusters in several adjacent loops of Sp?tzle far from the cystine-knot structural domain. We designed six mutants of Sp?tzle based on the evolutionary epitope and transfected them into a stable cell line expressing the luciferase reporter gene under the control of the drosomycin promoter. Luciferase assays showed that these mutants cannot significantly activate the drosomycin promoter, suggesting the involvement of these sites in binding of Sp?tzle to the Toll receptor. These data highlight the importance of the Trp-loop of the mushroom-shaped Sp?tzle dimer in Toll receptor activation and demonstrate that evolution-guided site-specific mutagenesis represents a useful and promising strategy for understanding the ligand-receptor interaction. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Received 14 January 2009; received after revision 14 February 2009; accepted 09 March 2009     

Abnormalities of meioses in male reciprocal hybrids between the Hiroshima and Ichinoseki populations ofRana japonica

In order to elucidate cytogenetically the cause of male sterility in intraspecific hybrids ofRana japonica, the behavior of chromosomes in the first meiosis was observed in spermatocytes from male reciprocal hybrids between two populations from Hiroshima and Ichinoseki. In the parental Hiroshima and Ichioseki populations, 2530 (96.7%) meiotic spreads had 13 bivalents and 78 (3.0%) contained 12 bivalents and two univalents, whereas in reciprocal hybrids only 337 (7.0%) contained 13 bivalents and the other 4445 (93.0%) had 2–26 univalents. A total of 31647 (93.4%) bivalents was ring-shaped and the other 2234 (6.6%) were rod-shaped in both parental populations, whereas in reciprocal hybrids 26352 (57.1%) and 19819 (42.9%) bivalents were ring- and rod-shaped, respectively. These results show that meiotic chromosomes of reciprocal hybrids are characterized by a remarkable increase in univalents and rod-shaped bivalents.     

Hydroxyproline-rich glycoproteins in plant reproductive tissues: structure, functions and regulation

The plant reproductive process of pollination involves a series of interactions between the male gametophyte (the pollen grain or pollen tube) and extracellular matrix (ECM) molecules secreted by different cell types along the pollen tube growth pathway in the female organ, the pistil. These interactions are believed to signal and regulate the pollen tube growth process to effect successful delivery of the sperm cells to the ovules where fertilization takes place. Hydroxyproline-rich glycoproteins secreted by plant cells are believed to play a broad range of functions, ranging from providing structural integrity to mediating cell-cell interactions and communication. The pistil and pollen tube ECM is enriched in these highly glycosylated proteins. Our discussions here will focus on a number of these proteins for which most information has been available, from Nicotiana tabacum, its self-incompatible relative N. alata, and Zea mays. In addition, the regulation of the synthesis and glyco-modification of one of these proteins, TTS (transmitting tissue-specific) protein from N. tabacum will be discussed in the light of how differential glycosylation may be used to regulate molecular interactions within the ECM.     

The male pheromone of the old house borerHylotrupes bajulus (L.) (Coleoptera: Cerambycidae): Identification and female response

We report here the identification of the long-range, male-produced sex pheromone of the Old house borerHylotrupes bajulus. Chemical analysis of hexane extracts obtained by surface extraction from dissected prothoracic glands and from headspace samples of the two sexes, revealed male-specific compounds: (3R)-3-hydroxy-2-hexanone, 2-hydroxy-3-hexanone, the diastereomeric diols (2R, 3R)-2,3-hexanediol and (2S, 3R)-2,3-hexanediol, 2,3-hexanedione, as well as 1-butanol.In wind tunnel bioassays we tested the influence of these male-specific compounds from the prothoracal glands on the behaviour of unmated and mated females. Specific behavioural sequences of the tested females (activity, running behaviour, searching, cleaning, flying, extension of ovipositor) were recorded. Unmated females were attracted by male beetles, headspace extracts of males, synthetic blends of the major pheromone compounds as well as by the components (3R)-3-hydroxy-2-hexanone, and the diastereomeric diols. Hexane, female beetles and 2,3-hexanedione did not attract unmated females. The reactions of mated females to male beetles and headspace samples did not differ significantly from those of the controls.The results of the bioassays show that the two-stage premating behaviour is initiated by emission of a long-range sex pheromone from the male prothoracal glands, which functions as an activator, attractant, and possibly aphrodisiac for unmated females.     

Fluorescence replication banding of frog chromosomes

To identify individual chromosomes of a frog karyotype by their fluorescence banding patterns, chromosomes were stained with actinomycin D and 4,6-diamidino-2-phenylindole (DAPI) after incorporation of BrdU during the late S-phase. The chromosomes of three Rana species which were selected for this study (R. ridibunda, R. lessonae and R. japonica) showed well-defined late replication bands. The fluorescence patterns obtained were the reverse of those produced by a 4Na-EDTA Giemsa-staining technique. Fluorescence patterns of the two water frog species (R. ridibunda and R. lessonae) were similar to each other, except for the different fluorescence of the centromeric heterochromatin, which gave extremely bright signals in R. ridibunda but no signal in R. lessonae. Experiments also showed differences between the fluorescence patterns of R. lessonae chromosome 13 in the Italian and Luxembourgian populations. These results sho w that the fluorescence replication banding using actinomycin D and DAPI is very effective in identifying individual frog chromosomes and detecting their structural changes. Received 7 June 1996; received after revision 23 July 1996; accepted 21 August 1996     

Physiological role of apyrene spermatozoa ofBombyx mori

Summary Observation by electron and phase-contrast microscopy demonstrated that inBombyx mori the dissociation of eupyrene bundles, apparently through digestion of the prostatic secretion or an endopeptidase, was promoted by the vigorously flagellating movement of apyrene spermatozoa in the spermatophore.     

The oak processionary caterpillar (Thaumetopoea processionea L.) an urticating caterpillar related to the pine processionary caterpillar (Thaumetopoea pityocampa Schiff.) (Lepidoptera,Thaumetopoeidae)

Summary The hairs ofThaumetopoea processionea caterpillars (Lepidoptera) provoke a cutaneous reaction in man and animals. The urticating apparatus, the urticating gland which produces hairs, and the urticating hairs, are similar to those of theT. pityocampa caterpillar. The irritant fraction extracted from hairs contains soluble proteins; one of these shows immunological identity with thaumetopoein, the urticating protein of theTh. pityocampa caterpillar. This thaumetopoein-like protein is currently undergoing isolation and will be subjected to dermatological tests.     

Ultrasonic vocalizations by adult prairie voles,Microtus ochrogaster

Summary Male and femaleMicrotus ochrogaster were presented with anesthetized and awake conspecifics while ultrasonic vocalizations (USVs) were monitored. Males produced significantly more USVs than females during 5-min testing sessions. Males tended to produce more USVs to unfamiliar females than to familiar female siblings. Sexual experience led to increased USV scores by males. These results suggest that USVs by male prairie voles communicate to females the male's gender and his availability for reproductive behavior.     

Symbionts in the female tsetse flyGlossina morsitans morsitans

Summary A rickettsia-like symbiont, located in the ovaries ofG. m. morsitans is maternally transmitted to the offspring via the egg. It is suggested that they may be essential for normal ovarian development.

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Résumé Un symbionte de type rickettsien localisé dans les ovaires deG. m. morsitans est transmis maternellement par l'uf à la progéniture. Le symbionte est probablement indispensable au développement normal des ovaires.

     

Mixed life cycles in the transitional zone between voltinisms in the fall webworm,Hyphantria cunea

The fall webworm,Hyphantria cunea Drury, was introduced from North America into Japan at half a century ago. At present, the populations north of 36° N are bivoltine and those south of 36° N are trivoltine. In the present study, the life cycle in the transitional zone between voltinisms was clarified in Tsukuba (36.1° N). When fourth instar larvae of the second generation were collected in the field and maintained in the laboratory, the incidence of pupal diapause increased when the sampling date was delayed from 16 August to 5 September. The fact that some developed without diapause inidcates the occurrence of a trivoltine life cycle in this locality, where a bivoltine life cycle is predominant. The critical photoperiod for diapause induction at 20 and 25°C was 14 h 13 min and 14 h 10 min, respectively. The photoperiodic response may explain the rapid increase in diapause incidence in late summer. In laboratory-reared diapause pupae, the time and period of chilling (5°C) greatly influenced the time required for adult emergence at the final incubation temperature of 25°C. However, diapause pupae chilled for a sufficiently long period developed to adulthood rapidly, irrespective of the conditions before chilling. Therefore, the difference in timing of entering diapause between pupae of the second and third generations would not result in their temporal reproductive isolation in the following spring. It is concluded that the mixed voltinism in Tsukuba is not a result of a mixture of genetically distinct bivoltine and trivoltine populations, but a result of the phenotypic plasticity in a genetically rather homogeneous population.