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.     

High affinity recognition of a <Emphasis Type="Italic">Phytophthora</Emphasis> protein by <Emphasis Type="Italic">Arabidopsis</Emphasis> via an RGD motif

The RGD tripeptide sequence, a cell adhesion motif present in several extracellular matrix proteins of mammalians, is involved in numerous plant processes. In plant-pathogen interactions, the RGD motif is believed to reduce plant defence responses by disrupting adhesions between the cell wall and plasma membrane. Photoaffinity cross-linking of [¹²⁵I]-azido-RGD heptapeptide in the presence of purified plasma membrane vesicles of Arabidopsis thaliana led to label incorporation into a single protein with an apparent molecular mass of 80 kDa. Incorporation could be prevented by excess RGD peptides, but also by the IPI-O protein, an RGD-containing protein secreted by the oomycete plant pathogen Phytophthora infestans. Hydrophobic cluster analysis revealed that the RGD motif of IPI-O (positions 53–56) is readily accessible for interactions. Single amino acid mutations in the RGD motif in IPI-O (of Asp⁵⁶ into Glu or Ala) resulted in the loss of protection of the 80-kDa protein from labelling. Thus, the interaction between the two proteins is mediated through RGD recognition and the 80-kDa RGD-binding protein has the characteristics of a receptor for IPI-O. The IPI-O protein also disrupted cell wall-plasma membrane adhesions in plasmolysed A. thaliana cells, whereas IPI-O proteins mutated in the RGD motif (D56A and D56E) did not.Received 23 October 2003; received after revision 5 December 2003; accepted 12 December 2003     

Can we predict the mating pattern ofDrosophila females from the sperm length distribution in males?

Summary In order to test the validity of the prediction of the mating pattern of females from the sperm length distribution in males, three species ofDrosophila were analysed. Males in the three species are equally polygynous but females differ in the level of polyandry. A low recurrence polyandry is observed in the sperm dimorphic speciesD. affinis while a high recurrence polyandry is observed in the sperm monomorphic speciesD. latifasciaeformis andD. littoralis. These results are consistent with the hypothesis proposed previously that sperm dimorphism in males can only be maintained by a selective alternative in females (i.e. facultative female polygamy), whereas a stricter mating system (e.g., obligatory polyandry) should only result in sperm monomorphism irrespective of the absolute value of sperm length.     

New insights into the molecular mechanisms of sperm-egg interaction

At the moment of insemination millions of mammalian sperm cells are released into the female reproductive tract in order to find a single cell – the oocyte. The spermatozoa subsequently ignore the thousands of cells they make contact with during their journey to the site of fertilisation, until they reach the surface of the oocyte. At this point, they bind tenaciously to the acellular coat, known as the zona pellucida, that surrounds the oocyte and initiate the chain of cellular interactions that will culminate in fertilization. These exquisitely cell- and species-specific recognition events are among the most strategically important cellular interactions in biology. Understanding the cellular and molecular mechanisms that underpin them has implications for diagnosis of the aetiology of human infertility and the development of novel targets for fertility regulation. Herein, we describe two models indicating the plethora of highly orchestrated molecular interactions underlying successful sperm zona binding and sperm oocyte fusion. Received 17 December 2006; received after revision 31 January 2007; accepted 16 March 2007     

Pheromone behavioral responses in unusual male European corn borer hybrid progeny not correlated to electrophysiological phenotypes of their pheromone-specific antennal neurons

In genetic studies on the sex pheromone communication system of two races of European corn borer, which use opposite pheromone blends of the E and Z compounds, it was found that antennal olfactory cell response amplitudes to the two compounds were controlled by an autosomal factor, whereas behavioral responses to the blends were controlled by a sex-linked locus. Because of the difference in genetic controls, it was postulated that some unusual males would be produced in F₂ crosses between these two races. These unusual males would have antennal olfactory cells that respond as the Z-race males, but would respond behaviorally to the E blend. The present studies combined behavioral studies in a flight tunnel and single cell electrophysiological studies to show that these unusual males do indeed exist. These findings show that the spike amplitude of peripheral olfactory cells is not important in regulating species- or race-specific pheromone responses, as compared to some central nervous system factor assesses the spike frequencies from different pheromone-component-specific cells on the antenna. This factor seems to be essential in governing the pheromone-blend specific behavioral responses of male moths.     

Can we predict the mating pattern of Drosophila females from the sperm length distribution in males?

In order to test the validity of the prediction of the mating pattern of females from the sperm length distribution in males, three species of Drosophila were analysed. Males in the three species are equally polygynous but females differ in the level of polyandry. A 'low recurrence polyandry' is observed in the sperm dimorphic species D. affinis while a 'high recurrence polyandry' is observed in the sperm monomorphic species D. latifasciaeformis and D. littoralis. These results are consistent with the hypothesis proposed previously that sperm dimorphism in males can only be maintained by a selective alternative in females (i.e. facultative female polygamy), whereas a stricter mating system (e.g. 'obligatory' polyandry) should only result in sperm monomorphism irrespective of the absolute value of sperm length.     

Disruption of sperm release from insect testes by cytochalasin and β-actin mRNA mediated interference

Release of sperm bundles from moth testes is controlled by the local circadian oscillator. The mechanism which restricts migration of sperm bundles to a few hours each day is not understood. We demonstrate that a daily cycle of sperm release is initiated by the migration of folded apyrene sperm bundles through a cellular barrier at the testis base. These bundles have conspicuous concentrations of actin filaments at their proximal end. Inhibition of actin polymerization by cytochalasin at a specific time of day inhibited sperm release from the testis. Likewise, application of double-stranded actin RNA specifically inhibited sperm release. This RNA-mediated interference (RNAi) lowered the pool of actin mRNA in tissues involved in sperm release. The decline in mRNA levels resulted in the selective depletion of F-actin from the tip of apyrene sperm bundles, suggesting that this actin may be involved in the initiation of sperm release. Combined results of RNAi experiments at physiological, cellular and molecular levels identified unique cells that are critically involved in the mechanism of sperm release.Received 11 April 2003; received after revision 2 May 2003; accepted 27 May 2003     

Cbl signaling networks in the regulation of cell function

Cbl proteins control multiple cellular processes by acting as ubiquitin ligases and multifunctional adaptor molecules. They are involved in the control of cell proliferation, differentiation and cell morphology, as well as in pathologies such as autoimmune diseases, inflammation and cancer. Here we review recent advances in understanding the role of Cbl and the importance of a growing repertoire of Cbl-interacting proteins in the regulation of signaling pathways triggered by growth factors, antigens, cell adhesion, cytokines and hormones. We also address key issues of the nature of proteins that bind Cbl in particular cells, where they are located, and how they are altered or traffic within cells upon stimulation. It is becoming obvious that temporal and spatial changes in Cbl signaling networks are essential for the control of physiological processes in a variety of cells and organs and that their deregulation can result in the development of human diseases.Received 22 January 2003; received after revision 11 March 2003; accepted 26 March 2003     

Male production induced by antibiotic treatment inEncarsia formosa (Hymenoptera: Aphelinidae), an asexual species

The production of large numbers of males in the thelytokous speciesEncarsia formosa was induced by feeding antibiotics to their mothers. The males induced by antibiotic treatment produce sperm and sometimes mate with females, but insemination does not occur.     

Soluble proteins of chemical communication in the social wasp <Emphasis Type="Italic">Polistes dominulus</Emphasis>

Members of the odorant-binding protein (OBP) and chemosensory protein (CSP) families were identified and characterised in the sensory tissues of the social wasp Polistes dominulus (Hymenoptera: Vespidae). Unlike most insects so far investigated, OBPs were detected in antennae, legs and wings, while CSPs appeared to be preferentially expressed in the antennae. The OBP is very different from the homologous proteins of other Hymenopteran species, with around 20% of identical residues, while the CSP appears to be much better conserved. Both OBP and CSP, not showing other post-translational modifications apart from disulphide bridges, were expressed with high yields in a bacterial system. Cysteine pairing in the recombinant and native proteins follows the classical arrangements described for other members of these classes of proteins. OBPs isolated from the wings were found to be associated with a number of long-chain aliphatic amides and other small organic molecules. Binding of these ligands and other related compounds was measured for both recombinant OBP and CSP.Received 14 May 2003; received after revision 8 June 2003; accepted 12 June 2003     

Glycoconjugates in sperm function and gamete interactions: how much sugar does it take to sweet-talk the egg?

Glycoconjugates in the mammalian reproductive tract are critical components of the molecular mechanisms that control sperm maturation, sperm transport and gamete interactions. In the oviduct of many species, sperm transport and maturation are regulated by protein-carbohydrate interactions that form a sperm reservoir. Subsequently, gamete interactions are mediated by the binding of lectin-like sperm proteins with carbohydrate moieties on the zona pellucida. The sperm glycocalyx is extensively modified during sperm transport and maturation. Multiple functions have been proposed for this dense carbohydrate layer overlying the sperm plasmalemma, and sperm-surface carbohydrates have been implicated in immune-mediated human infertility. The structure and function of glycoconjugates in the oviductal sperm reservoir, the zona pellucida, and on the sperm surface are reviewed.     

Aptamer and its applications in neurodegenerative diseases

Aptamers are small single-stranded DNA or RNA oligonucleotide fragments or small peptides, which can bind to targets by high affinity and specificity. Because aptamers are specific, non-immunogenic and non-toxic, they are ideal materials for clinical applications. Neurodegenerative disorders are ravaging the lives of patients. Even though the mechanism of these diseases is still elusive, they are mainly characterized by the accumulation of misfolded proteins in the central nervous system. So it is essential to develop potential measures to slow down or prevent the onset of these diseases. With the advancements of the technologies, aptamers have opened up new areas in this research field. Aptamers could bind with these related target proteins to interrupt their accumulation, subsequently blocking or preventing the process of neurodegenerative diseases. This review presents recent advances in the aptamer generation and its merits and limitations, with emphasis on its applications in neurodegenerative diseases including Alzheimer’s disease, Parkinson’s disease, transmissible spongiform encephalopathy, Huntington’s disease and multiple sclerosis.     

Sperm predence in a hermaphroditic nematode (Caenorhabditis elegans) is due to competitive superiority of male sperm

When male and hermaphroditeCaenorhabditis elegans mate, the male's sperm outcompete the hermaphrodite's own sperm and fertilize a majority of the offspring. Here, we investigate the mechanism of male sperm precedence. We rule out the possibility that male sperm are stronger and more competitive because they are activated later than hermaphrodite sperm. We also find that a previously known gender difference in sperm activation does not influence sperm competition. Male sperm, rinsed free of seminal fluid, retained the capacity to take precedence after artificial insemination. Therefore, we conclude that male sperm themselves are competitively superior to hermaphrodite sperm. This trait maximizes outcrossing after mating and may increase both genetic diversity and heterozygosity of offspring whose parents, due to self-fertilization, may be highly homozygous.     

Entwicklungsleistungen transplantierter Genital- und Analanlagen vonMusca domestica undPhormia regina

Summary InDrosophila, both sexes develop their whole genital and anal apparatus from one single median genital disc. Larvae ofMusca domestica andPhormia regina are equipped with 3 post-abdominal discs. In females, the median genital disc develops into the external analia, the hind gut and the parovaria. The 2 lateral genital discs together yield the chitinized structures of the 8th abdominal segment and the whole internal genital apparatus except the parovaria and the gonads. When transplanted into larval hosts of the same age as the donors, each lateral genital disc gives rise to an almost complete internal genital apparatus, i.e. they show a regulative capacity without previously being cultured. The entire male genital and anal apparatus develops from the median genital disc. The lateral genital discs of the male, as revealed by transplantation and exstirpation, seem to take part in the development of the syntergite 7+8.     

Sperm enzyme activity analysis in individual sperm for detection of heritable mutations in mammals

Summary Male mice were injected i.p. with 2.5 mg/kg mitomycin C, 100 mg/kg ethyl nitrosourea or saline and mated with untreated virgin females five weeks later. Sperm from 64 of the F₁ male progeny were analyzed histochemically for acrosin, succinic dehydrogenase and alpha-glycerophosphate dehydrogenase activity. The frequency of F₁ males with sub-normal sperm enzyme activity was significantly higher among progeny from treated males than in controls. These results show that analysis of sperm enzyme activity in F₁ males is a practical method for detection of transmitted mutations induced in a treated parent.We gratefully acknowledge USPHS, NIEHS grant 1 RO1 ES02607-02 and technical assistance by G. M. Oldford.     

Are elicitins cryptograms in plant-Oomycete communications?

Stimulation of plant natural defenses is an important challenge in phytoprotection prospects. In that context, elicitins, which are small proteins secreted by Phytophthora and Pythium species, have been shown to induce a hypersensitive-like reaction in tobacco plants. Moreover, these plants become resistant to their pathogens, and thus this interaction constitutes an excellent model to investigate the signaling pathways leading to plant resistance. However, most plants are not reactive to elicitins, although they possess the functional signaling pathways involved in tobacco responses to elicitin. The understanding of factors involved in this reactivity is needed to develop agronomic applications. In this review, it is proposed that elicitins could interact with regulating cell wall proteins before they reach the plasma membrane. Consequently, the plant reactivity or nonreactivity status could result from the equilibrium reached during this interaction. The possibility of overexpressing the elicitins directly from genomic DNA in Pichia pastoris allows site-directed mutagenesis experiments and structure/function studies. The recent discovery of the sterol carrier activity of elicitins brings a new insight on their molecular activity. This constitutes a crucial property, since the formation of a sterol-elicitin complex is required to trigger the biological responses of tobacco cells and plants. Only the elicitins loaded with a sterol are able to bind to their plasmalemma receptor, which is assumed to be an allosteric calcium channel. Moreover, Phytophthora and Pythium do not synthesize the sterols required for their growth and their fructification, and elicitins may act as shuttles trapping the sterols from the host plants. Sequence analysis of elicitin genes from several Phytophthora species sheds unexpected light on the phylogenetic relationships among the genus, and suggests that the expression of elicitins is under tight regulatory control. Finally, general involvement of these lipid transfer proteins in the biology of Pythiaceae, and in plant defense responses, is discussed. A possible scheme for the coevolution between Phytophthora and tobacco plants is approached.     

Schistosoma mansoni: A chemoattractive factor released by males and its receptor in females

Chemoattraction between developing (23- to 28-day-old) and adult (10- to 12-week-old)Schistosoma mansoni males and females has been described previously. We report here on in vitro attraction of mature worms to released products of worms of the opposite sex. In the absence of a stimulus worms migrated randomly and did not show any preference in their orientation. Males were not significantly attracted to released products of females. Females exhibited greatest attraction to released products of males, and some attraction to the lipid fraction of released products of males. The aqueous fraction of male-released products slightly repelled females. Prior exposure to released products of males rendered females unresponsive to subsequent exposure, suggesting receptor saturation. These findings have important potential implicatons for the control of schistosomiasis by intercepting attraction and mating between males and females.     

The role of assortative mating among age-classes inDrosophila melanogaster: preliminary results

Summary A series of experiments on age-related mating success and productivity provides evidence for assortative mating among three out of four age-classes inD. melanogaster. The preferred mating does not always result in the highest productivity. Three age classes of males contribute to reproduction while only females of the youngest age-class are involved. The progeny size is more affected by the age of the females than that of males. It is assumed that these findings must have important implications for generation overlap in natural populations.     

Infection of frog tadpoles (Amphibia) by insect parasitic nematodes (Rhabditida)

Summary Infective stage juveniles ofNeoaplectana carpocapsae (Steinernematidae) andHeterohabditis heliothidis (Heterorhabditidae) were able to penetrate through the alimentary tract of young tadpoles ofHyla regilla (Hylidae) andXenopus laevis (Pipidae) and enter the body cavity. Some infectives ofN. carpocapsae were able to release their symbiotic bacterium,Xenorhabdus nematophilus inside the host and in two cases, the nematodes developed into adult females before they perished. Tadpole mortality was associated with foreign bacteria entering the penetration holes made by the invading nematodes. The infective stage juveniles of both nematodes frequently encountered a host defense reaction upon reaching the tadpole's infective stage juveniles of both nematodes frequently encountered a host defense reaction upon reaching the tadpole's coelom.