Female Accessory Glands Research Articles

Gonadal Transcriptomic Analysis Reveals Novel Sex-Related Genes in Bactrocera dorsalis.

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) is one of the most devastating agricultural pests worldwide due to its high reproductive and invasive abilities. The elucidation of its gonadal developmental characteristics and the identification of sex-related genes will provide a useful genetic basis for reproductive-based pest control. Here, the gonadal transcriptome of B. dorsalis was sequenced, and novel gonad-specific expressed genes were analyzed. A total of 1338, 336, 35, and 479 differentially expressed genes (DEGs) were found in the testis (TE), ovary (OV), female accessory gland (FAG), and male accessory gland (MAG), respectively. Furthermore, 463 highly expressed gonad-specific genes were identified, with the TE having the highest number of specific highly expressed genes, at 402, followed by 51 in the OV, 9 in the MAG, and only 1 in the FAG. Strikingly, approximately half of highly expressed gonad-specific genes were uncharacterized. Then, it was found that 35, 17, 3, 2, and 1 of 202 uncharacterized highly expressed TE-specific genes encoded proteins that contained transmembrane domains, signal peptides, high-mobility group boxes, the zinc finger domain, and the BTB/POZ domain, respectively. Interestingly, approximately 40% of uncharacterized highly expressed gonad-specific genes encoding proteins were not predicted to possess functional motifs or domains. Finally, the spatiotemporal expression and sequence characterization of six novel highly expressed gonad-specific genes were analyzed. Altogether, our findings provide a valuable dataset for future functional analyses of sex-related genes and potential target sites for pest control.

Identification and characterization of uridine diphosphate glycosyltransferases in Dioryctria abietella, with an emphasis on putative roles in olfaction and reproduction

AbstractMultiple physiological processes are involved in the interactions between the herbivore Dioryctria abietella and Pinaceae plants, with a key adaptation being the metabolic detoxification of host plant defensive substances. Moreover, the synthetic insecticides applied to control this coneworm are also shaping genes related to detoxification, such as the uridine diphosphate (UDP) glycosyltransferase (UGT) genes. Yet, the molecular mechanisms underpinning the resistance of D. abietella to toxic compounds remain unknown. In this study, we present an important UGT gene family involved in the detoxification mechanisms of D. abietella. Combining bioinformatic and transcriptomic approaches, a total of 37 UGT‐coding genes were identified from the transcriptome of D. abietella, with 20 full‐length sequences that shared high homology with UGTs found in other pyralid moths. These DabiUGTs were phylogenetically clustered into 12 subfamilies, with the three small clusters in the UGT33 and UGT40 clades mainly contributing to the size of the UGT gene repertoire in D. abietella. Expression profiles obtained from RNA sequencing (RNA‐Seq), reverse transcription PCR (RT‐PCR) and quantitative real‐time PCR (qPCR) revealed that over half of DabiUGTs had a broad tissue expression profile, with 28, 28, 31 and 35 genes detected separately in antennae, legs, abdomens and reproductive tissues. Notably, DabiUGT20, DabiUGT25, DabiUGT28 and DabiUGT34 were all significantly enriched in the antennae. Several DabiUGTs had significant expression levels in reproductive tissues, including DabiUGT6, DabiUGT9, DabiUGT24, DabiUGT26 and DabiUGT32 in female accessory glands, DabiUGT15 in male accessory glands and DabiUGT25 in male testes. Altogether, our results identify candidate DabiUGTs for mediating olfaction and reproduction in D. abietella, warranting further investigation into the roles of DabiUGTs in pesticide resistance, odorant detection and reproduction.

Female accessory glands of Adoxophyes honmai contain elicitor inducing tea leaves to arrest the egg-larval parasitoid, Ascogaster reticulata

Female accessory glands of Adoxophyes honmai contain elicitor inducing tea leaves to arrest the egg-larval parasitoid, Ascogaster reticulata

Morphology and ultrastructure of the colleterial glands in Myzinum flower wasps (Tiphiidae, Hymenoptera)

The strategies used by females to ensure the survival of their offspring are one of the key elements in insect reproductive biology. Ectoparasitoid hymenopterans, such as tiphiid wasps, use their ovipositor to deliver a single egg on the host body, where the larva develops. Here we describe the colleterial glands in females of two species of Myzinum and discuss the homology with other glands among Hymenoptera. The glands appear as two large opaque round sac-like structures that are connected to the oviduct. Histological sections revealed a compound gland, with a single layer of class-1 epithelial cells surrounded by a layer of class-3 gland cells, which is uncommon in Hymenoptera. Ultrastructure points to active protein synthesis, consistent with the role of colleterial glands. Despite the consensus that colleterial glands in Hymenoptera evolved with two distinct forms, the venom gland and Dufour’s gland, the three glands were present in both Myzinum species. The few studies regarding the structure, function, and homology among the female accessory glands in Hymenoptera do not allow to make conclusive statements about the evolution of these glands. Therefore, the finding in Myzinum is probably the reflection of a lack of information, rather than an exception within Aculeata.

Male-derived transcripts isolated from the mated female reproductive tract in Drosophila melanogaster.

In species with internal fertilization, sperm, and seminal fluid are transferred from male to female during mating. While both sperm and seminal fluid contain various types of molecules, including RNA, the role of most of these molecules in the coordination of fertilization or in other possible functions is poorly understood. In Drosophila, exosomes from the accessory gland, which produces seminal fluid, are transferred to females, but their potential cargoes have not been described. Moreover, while the RNA composition of sperm has been described in several mammalian species, little work on this problem has occurred in Drosophila. Here we use single nucleotide polymorphism differences between males and females from a set of highly inbred lines of D. melanogaster, and transcriptome data from the female reproductive tract, sperm, testis, and accessory gland, to investigate the potential origin, male vs female, RNA molecules isolated from 3 female reproductive tract organs, the seminal receptacle and spermatheca, which store sperm, and the parovaria, which does not. We find that mated females carry male-derived transcripts from many genes, including those that are markers of the accessory gland and known seminal fluid proteins. Our observations also support the idea that intact sperm transcripts can be isolated from the female sperm storage organs.

Reproductive Biology of Muga Silk Moth <i>Antheraea assamensis</i> Helfer

Results of morphological and morphometric study of male and female reproductive systems of Antheraea assamensis Helfer (Saturniidae: Lepidoptera), a silk moth cultivated exclusively in the North Eastern Region of India are reported herein along with some observations on its reproductive performance in four seasons. The male reproductive system consists of a pair of testes, paired vasa deferentia, seminal vesicle, ductus ejaculatorius duplex, ductus ejaculatorius simplex, an aedeagus and a pair of male accessory glands. The female reproductive system consists of a pair of ovaries, each with four ovarioles, paired lateral oviducts, a common oviduct which has a dorsal protrusion, the vestibulum, a bursa copulatrix, a spermatheca and a pair of female accessory glands with their reservoirs. Fecundity has been found to be positively correlated with female pupal weight in each season. Hatching % of eggs has been found lowest and significantly different (p<0.001) in summer as compared to autumn, winter and spring seasons. Morphology of eupyrene and apyrene sperm bundles and sperm has also been reported.

Female factors modulate Sex Peptide’s association with sperm in Drosophila melanogaster

BackgroundMale-derived seminal fluid proteins (SFPs) that enter female fruitflies during mating induce a myriad of physiological and behavioral changes, optimizing fertility of the mating pair. Some post-mating changes in female Drosophila melanogaster persist for ~10–14 days. Their long-term persistence is because the seminal protein that induces these particular changes, the Sex Peptide (SP), is retained long term in females by binding to sperm, with gradual release of its active domain from sperm. Several other “long-term response SFPs” (LTR-SFPs) “prime” the binding of SP to sperm. Whether female factors play a role in this process is unknown, though it is important to study both sexes for a comprehensive physiological understanding of SFP/sperm interactions and for consideration in models of sexual conflict.ResultsWe report here that sperm in male ejaculates bind SP more weakly than sperm that have entered females. Moreover, we show that the amount of SP, and other SFPs, bound to sperm increases with time and transit of individual seminal proteins within the female reproductive tract (FRT). Thus, female contributions are needed for maximal and appropriate binding of SP, and other SFPs, to sperm. Towards understanding the source of female molecular contributions, we ablated spermathecal secretory cells (SSCs) and/or parovaria (female accessory glands), which contribute secretory proteins to the FRT. We found no dramatic change in the initial levels of SP bound to sperm stored in mated females with ablated or defective SSCs and/or parovaria, indicating that female molecules that facilitate the binding of SP to sperm are not uniquely derived from SSCs and parovaria. However, we observed higher levels of SP (and sperm) retention long term in females whose SSCs and parovaria had been ablated, indicating secretions from these female tissues are necessary for the gradual release of Sex Peptide’s active region from stored sperm.ConclusionThis study reveals that the SP-sperm binding pathway is not entirely male-derived and that female contributions are needed to regulate the levels of SP associated with sperm stored in their storage sites.

Functions of Egg-Coating Substances Secreted by Female Accessory Glands in Alderflies, Fishflies and Dobsonflies (Megaloptera).

Simple SummaryEgg masses of the insect order Megaloptera are coated with female accessory gland substances and are diverse in shape, color, and surface structure. Female alderflies (family Sialidae) and fishflies (family Corydalidae, subfamily Chauliodinae) lay eggs in a single layer on the substrate (although the eggs are occasionally multi-layered in the fishfly genus Nigronia), and the accessory gland substances are usually brown. In contrast, dobsonflies (Corydalidae, Corydalinae) lay a multi-layered hemispherical egg mass, and the accessory gland substances differ in color and chemical properties among genera and species. Egg masses of the dobsonfly genus Protohermes are coated with a sticky substance, while those of other dobsonflies are covered with hardened white powders, both of which contribute to the prevention of egg desiccation. The accessory gland substances of most species of Megaloptera also function to protect eggs from attack by oophagous predators, such as ladybird beetles and ants. The information on the parasitoids of Megaloptera egg masses are also discussed based on the present and previously reported observations.Eggs of insects are immobile and must endure harsh environmental conditions (e.g., low temperatures in winter and aridity in summer) and avoid attack by egg-eating predators, egg parasites, and microbes. Females of Megaloptera lay their eggs as a single- or multi-layered egg mass, which is coated with chemical substances secreted from the female reproductive accessory glands. In this study, we observed the egg masses laid by females of two species of Sialidae (alderflies), nine species of Chauliodinae (fishflies), and 23 species of Corydalinae (dobsonflies) belonging to the order Megaloptera and examined the functions of accessory gland substances coating the laid eggs. The female accessory gland is a single tube in alderflies and fishflies but a paired pouch in dobsonflies. The amount and color of the gland substances differ greatly among species. These substances prevent egg desiccation, inhibit egg feeding by ladybird beetles, and repel ants. Most characteristics of the egg mass structures and the effectiveness of accessory gland substances reflect the phylogeny of Megaloptera, although some differ among closely related taxa.

Female accessory gland fluid promotes sperm survival in yellow dung flies

Female and male reproductive traits co-evolve through pre- and post-copulatory sexual selection and sexual conflict. Although males typically transfer many sperm during copulation, only a small proportion reach the fertilization site because females often actively or passively reduce sperm number in their reproductive tract. Males may transfer accessory substances to protect their ejaculates against female selective processes, which benefits males but can harm females. In turn, females may use accessory gland fluids to control paternity or sperm storage. Female yellow dung flies (Scathophaga stercoraria) have paired accessory glands that produce fluids involved in fertilization and egg laying. One proposed function for these fluids is spermicide. Alternatively, female accessory gland fluid may help keep sperm alive to avoid fertilization failure or encourage sperm competition. Using yellow dung flies, we investigated the interaction of female accessory gland fluid with sperm in vitro. Significantly more sperm remained alive when exposed to accessory gland fluid compared to buffer only (63% vs. 44%). We conclude that female accessory gland fluid in yellow dung flies can help nourish rather than kill male sperm, although selective nourishment of sperm is as consistent with cryptic female choice as is selective spermicide.

Reproductive Systems, Transfer and Digestion of Spermatophores in Two Asian Luciolinae Fireflies (Coleoptera: Lampyridae).

Simple SummaryDuring mating, fireflies may transfer sperm to the female wrapped in food materials (spermatophores). We investigate in two firefly species structures in both male and female that indicate production and receipt of spermatophores. Their structure, how they attach and discharge the sperm inside the female, and how long they persist might indicate if these fireflies mate only once or more. Potential differences between males and females of both fully winged and species with flightless females are determined. An overview of present knowledge of female reproductive anatomy is given. An argument is mounted suggesting direct observation of biological structures like spermatophores might be the only reliable way of determining their presence.The internal reproductive anatomy of males and females of two Asian Luciolinae fireflies Emeia pseudosauteri (Geisthardt, 2004) and Abscondita chinensis (L., 1767) is described, and the time course for spermatophore transfer and digestion examined. E. pseudosauteri is sexually dimorphic, with a flightless female, and Abs. chinensis is sexually monomorphic, with the female flighted. Both are monandrous. Possible female accessory glands are described for the first time for both species. An overview of present knowledge of female reproductive anatomy in the Luciolinae reveals males of 18 species in 10 genera may produce spermatophores and permits speculation about spermatophore production in another 16 genera.

Multi-level analysis of reproduction in an Antarctic midge identifies female and male accessory gland products that are altered by larval stress and impact progeny viability

The Antarctic midge, Belgica antarctica, is a wingless, non-biting midge endemic to Antarctica. Larval development requires at least 2 years, but adults live only 2 weeks. The nonfeeding adults mate in swarms and females die shortly after oviposition. Eggs are suspended in a gel of unknown composition that is expressed from the female accessory gland. This project characterizes molecular mechanisms underlying reproduction in this midge by examining differential gene expression in whole males, females, and larvae, as well as in male and female accessory glands. Functional studies were used to assess the role of the gel encasing the eggs, as well as the impact of stress on reproductive biology. RNA-seq analyses revealed sex- and development-specific gene sets along with those associated with the accessory glands. Proteomic analyses were used to define the composition of the egg-containing gel, which is generated during multiple developmental stages and derived from both the accessory gland and other female organs. Functional studies indicate the gel provides a larval food source as well as a buffer for thermal and dehydration stress. All of these function are critical to juvenile survival. Larval dehydration stress directly reduces production of storage proteins and key accessory gland components, a feature that impacts adult reproductive success. Modeling reveals that bouts of dehydration may have a significant impact on population growth. This work lays a foundation for further examination of reproduction in midges and provides new information related to general reproduction in dipterans. A key aspect of this work is that reproduction and stress dynamics, currently understudied in polar organisms, are likely to prove critical in determining how climate change will alter their survivability.

Proteome profiling reveals tissue-specific protein expression in male and female accessory glands of the silkworm, Bombyx mori.

Male accessory gland (MAG) and female accessory gland (FAG) of the reproductive system are, respectively, responsible for producing seminal proteins and adhesive proteins during copulation and ovulation. Seminal proteins are ejaculated to female along with sperms, whereas adhesive proteins are excreted along with eggs. Proteins from the male and female reproductive organs are usually indicative of rapid adaptive evolution. Understanding the reproductive isolation and species divergence requires identifying reproduction-related proteins from many different species. Here, we present our proteomic analyses of male and female accessory glands of the silkworm, Bombyx mori. Using LC/MS-MS, we identified 2133 MAG proteins and 1872 FAG proteins. In total, 652 proteins were significant more abundant in the MAG than in the FAG, including growth factors, odorant-binding proteins, enzymes, and proteins of unknown function. Growth factors and odorant-binding proteins are potential signaling molecules, whereas most of proteins of unknown function were found to be Lepidoptera-specific proteins with high evolutionary rates. Microarray experiments and semi-quantitative RT-PCR validated that MAG-specific proteins were expressed exclusively in male moths. Totally, 192 proteins were considered as FAG-specific proteins, including protease inhibitors, enzymes, and other proteins. Protease inhibitors were found to be the most abundant FAG-specific proteins, which may protect eggs from infection by inhibiting pathogen-derived proteases. These results provide comprehensive insights into copulation and oviposition. Moreover, the newly identified Lepidoptera-specific MAG proteins provide useful data for future research on the evolution of reproductive proteins in insects.

Molecular cloning, expression analysis and cellular localization of an LFRFamide gene in the cuttlefish Sepiella japonica

Neuropeptides are important regulators of physiological processes in metazoans, such as feeding, reproduction, and heart activities. In this study, an LFRFamide gene was identified from the cuttlefish Sepiella japonica (designated as SjLFRFamide). The full-length sequence of SjLFRFamide cDNA has 841bp, and the open reading frame contains 567bp encoding 188 amino acids, which shared high similarity with precursor SOFaRP2 from Sepia officinalis. The deduced SjLFRFamdie precursor protein contains a signal peptide and four different FLPs (FMRFamide-like peptides): one pentapeptide (TIFRFamide), two hexapeptides (NSLFRFamide and GNLFRFamide) and one heptapeptide (PHTPFRFamide). Multiple sequence alignment showed that SjLFRFamide contains rather conserved mature peptides, which all ended in FRF. The phylogenetic analysis suggests that SjLFRFamide belongs to the LFRFamide subfamily. The tissue distribution analysis through quantitative real-time PCR method showed that SjLFRFamide mRNA is significantly expressed in the brain, and slight trace are detected in female nidamental gland and accessory nidamental gland. In situ hybridization assay of the brain indicated that SjLFRFamide is transcribed in several different functional lobes, suggesting SjLFRFamide might associate with multiple physiological regulations, such as feeding, chromatophore regulation and reproduction. This is the first study describing LFRFamide in S. japonica, which might have great importance for cuttlefish artificial breeding.

How Egg Case Proteins Can Protect Cuttlefish Offspring?

Sepia officinalis egg protection is ensured by a complex capsule produced by the female accessory genital glands and the ink bag. Our study is focused on the proteins constituting the main egg case. De novo transcriptomes from female genital glands provided essential databases for protein identification. A proteomic approach in SDS-PAGE coupled with MS unveiled a new egg case protein family: SepECPs, for Sepia officinalis Egg Case Proteins. N-glycosylation was demonstrated by PAS staining SDS-PAGE gels. These glycoproteins are mainly produced in the main nidamental glands. SepECPs share high sequence homology, especially in the signal peptide and the three cysteine-rich domains. SepECPs have a high number of cysteines, with conserved motifs involved in 3D-structure. SDS-PAGE showed that SepECPs could form dimers; this result was confirmed by TEM observations, which also revealed a protein network. This network is similar to the capsule network, and it associates these structural proteins with polysaccharides, melanin and bacteria to form a tight mesh. Its hardness and elasticity provide physical protection to the embryo. In addition, SepECPs also have bacteriostatic antimicrobial activity on GRAM- bacteria. By observing the SepECP / Vibrio aestuarianus complex in SEM, we demonstrated the ability of these proteins to agglomerate bacteria and thus inhibit their growth. These original proteins identified from the outer egg case ensure the survival of the species by providing physical and chemical protection to the embryos released in the environment without any maternal protection.

Reproductive biology in Anophelinae mosquitoes (Diptera, Culicidae): Fine structure of the female accessory gland

The morphology and ultrastructure of female accessory reproductive glands of Anopheles maculipennis s.s., Anopheles labranchiae and Anopheles stephensi were investigated by light and electron microscopy. The reproductive system in these species is characterized by two ovaries, two lateral oviducts, a single spermatheca and a single accessory gland. The gland is globular and has a thin duct which empties into the vagina, near the opening of the spermathecal duct. Significant growth of the accessory reproductive gland is observed immediately after blood meal, but not at subsequent digestion steps. At ultrastructural level, the gland consists of functional glandular units belonging to type 3 ectodermal glands. The secretory cells are elongated and goblet shaped, with most of their cytoplasm and large nucleus in the basal part, close to the basement lamella. Finely fibrous electron-transparent material occupies the secretory cavity that is in contact with the end of a short efferent duct (ductule) emerging from the gland duct. The present study is the first detailed description of female accessory gland ultrastructure in Anophelinae and provides insights into the gland's functional role in the reproductive biology of these insects.

The molecular biology of the olive fly comes of age.

BackgroundOlive cultivation blends with the history of the Mediterranean countries since ancient times. Even today, activities around the olive tree constitute major engagements of several people in the countryside of both sides of the Mediterranean basin. The olive fly is, beyond doubt, the most destructive pest of cultivated olives. The female fly leaves its eggs in the olive fruit. Upon emergence, the larvae feed on the olive sap, thus destroying the fruit. If untreated, practically all olives get infected. The use of chemical insecticides constitutes the principal olive fly control approach. The Sterile Insect Technique (SIT), an environmentally friendly alternative control method, had been tried in pilot field applications in the 1970's, albeit with no practical success. This was mainly attributed to the low, non-antagonistic quality of the mixed-sex released insects. Many years of experience from successful SIT applications in related species, primarily the Mediterranean fruit fly, Ceratitis capitata, demonstrated that efficient SIT protocols require the availability of fundamental genetic and molecular information.ResultsAmong the primary systems whose understanding can contribute towards novel SIT approaches (or its recently developed alternative RIDL: Release of Insects carrying a Dominant Lethal) is the reproductive, since the ability to manipulate the reproductive system would directly affect the insect's fertility. In addition, the analysis of early embryonic promoters and apoptotic genes would provide tools that confer dominant early-embryonic lethality during mass-rearing. Here we report the identification of several genes involved in these systems through whole transcriptome analysis of female accessory glands (FAGs) and spermathecae, as well as male testes. Indeed, analysis of differentially expressed genes in these tissues revealed higher metabolic activity in testes than in FAGs/spermathecae. Furthermore, at least five olfactory-related genes were shown to be differentially expressed in the female and male reproductive systems analyzed. Finally, the expression profile of the embryonic serendipity-α locus and the pre-apoptotic head involution defective gene were analyzed during embryonic developmental stages.ConclusionsSeveral years of molecular studies on the olive fly can now be combined with new information from whole transcriptome analyses and lead to a deep understanding of the biology of this notorious insect pest. This is a prerequisite for the development of novel embryonic lethality female sexing strains for successful SIT efforts which, combined with improved mass-reared conditions, give new hope for efficient SIT applications for the olive fly.

The homeodomain protein ladybird late regulates synthesis of milk proteins during pregnancy in the tsetse fly (Glossina morsitans).

Regulation of tissue and development specific gene expression patterns underlies the functional specialization of organs in multi-cellular organisms. In the viviparous tsetse fly (Glossina), the female accessory gland is specialized to generate nutrients in the form of a milk-like secretion to support growth of intrauterine larva. Multiple milk protein genes are expressed specifically in the female accessory gland and are tightly linked with larval development. Disruption of milk protein synthesis deprives developing larvae of nutrients and results in extended larval development and/or in abortion. The ability to cause such a disruption could be utilized as a tsetse control strategy. Here we identify and delineate the regulatory sequence of a major milk protein gene (milk gland protein 1:mgp1) by utilizing a combination of molecular techniques in tsetse, Drosophila transgenics, transcriptomics and in silico sequence analyses. The function of this promoter is conserved between tsetse and Drosophila. In transgenic Drosophila the mgp1 promoter directs reporter gene expression in a tissue and stage specific manner orthologous to that of Glossina. Analysis of the minimal required regulatory region of mgp1, and the regulatory regions of other Glossina milk proteins identified putative homeodomain protein binding sites as the sole common feature. Annotation and expression analysis of Glossina homeodomain proteins identified ladybird late (lbl) as being accessory gland/fat body specific and differentially expressed between lactating/non-lactating flies. Knockdown of lbl in tsetse resulted in a significant reduction in transcript abundance of multiple milk protein genes and in a significant loss of fecundity. The role of Lbl in adult reproductive physiology is previously unknown. These results suggest that Lbl is part of a conserved reproductive regulatory system that could have implications beyond tsetse to other vector insects such as mosquitoes. This system is critical for tsetse fecundity and provides a potential target for development of a reproductive inhibitor.

Identification of pheromone-like compounds in male reproductive organs of the oriental locust Locusta migratoria

Despite the great economical interest of locusts in agriculture, knowledge on their chemoreception systems is still poor. Phenylacetonitrile is recognised as a pheromone of the desert locust Schistocerca gregaria, triggering gregarization, promoting aggregation and inhibiting courtship. However, in the other major locust species, Locusta migratoria, pheromones have not been reported. We have identified the two isomers of naphthylpropionitrile from the male reproductive organs of L. migratoria. Chemical synthesis has confirmed the identity of the two compounds. Both isomers show significant affinity to CSP91, a protein reported in the testis, but not to three other proteins of the same family (CSP180, CSP540 and CSP884) expressed in female accessory glands. The striking similarity of these compounds with phenylacetonitrile and the unusual nature of such chemicals strongly suggest that naphthylpropionitrile could be pheromones for L. migratoria, while their site of expression and binding activity indicate a role in communication between sexes.

Juvenile hormone and insulin suppress lipolysis between periods of lactation during tsetse fly pregnancy

Tsetse flies are viviparous insects that nurture a single intrauterine progeny per gonotrophic cycle. The developing larva is nourished by the lipid-rich, milk-like secretions from a modified female accessory gland (milk gland). An essential feature of the lactation process involves lipid mobilization for incorporation into the milk. In this study, we examined roles for juvenile hormone (JH) and insulin/IGF-like (IIS) signaling pathways during tsetse pregnancy. In particular, we examined the roles for these pathways in regulating lipid homeostasis during transitions between non-lactating (dry) and lactating periods. The dry period occurs over the course of oogenesis and embryogenesis, while the lactation period spans intrauterine larvigenesis. Genes involved in the JH and IIS pathways were upregulated during dry periods, correlating with lipid accumulation between bouts of lactation. RNAi suppression of Forkhead Box Sub Group O (FOXO) expression impaired lipolysis during tsetse lactation and reduced fecundity. Similar reduction of the JH receptor Methoprene tolerant (Met), but not its paralog germ cell expressed (gce), reduced lipid accumulation during dry periods, indicating functional divergence between Met and gce during tsetse reproduction. Reduced lipid levels following Met knockdown led to impaired fecundity due to inadequate fat reserves at the initiation of milk production. Both the application of the JH analog (JHA) methoprene and injection of insulin into lactating females increased stored lipids by suppressing lipolysis and reduced transcripts of lactation-specific genes, leading to elevated rates of larval abortion. To our knowledge, this study is the first to address the molecular physiology of JH and IIS in a viviparous insect, and specifically to provide a role for JH signaling through Met in the regulation of lipid metabolism during insect lactation.

Ultrastructure of female accessory glands in the scorpionfly Panorpa sexspinosa Cheng (Mecoptera: Panorpidae)

The histology and ultrastructure of the female accessory glands in the scorpionfly Panorpa sexspinosa Cheng was studied using light and transmission electron microscopy. The glands consist of a pair of distal elongate gland tubes and a basal common duct, which opens in the genital cavity at the dorsal side of the genital plate. The whole gland tubes and common duct are similar histologically and ultrastructurally. The epithelium of female accessory glands consists of two cell types: the outer secretory cells and the inner duct-forming cells. These two cells that join with a cuticular duct connecting to the inner intima constitute a functional glandular unit belonging to Class 3 glandular cells of epidermal glands. The secretory cells are rich in organelles, such as mitochondria, rough endoplasmic reticulum, and Golgi complex, indicating that they are active in secretion. The duct-forming cells are flattened with sparse-distributed organelles. These two kinds of cells are connected by septate junctions. The cuticular duct consists of a receiving and a conducting canal and is responsible for transferring the secretions of the secretory cell to the lumen. The receiving canal is formed of interrupted multilayered inner epicuticle and located in the irregular extracellular cavity of the secretory cell, bounded by microvilli. The conducting canal connects the inner intima and opens into the central lumen. The tentative functions of the secretions are briefly discussed.