Proteome dynamics in primary target organ of infectious bursal disease virus

Viruses induce dramatic changes in target tissue during pathogenesis, including host cellular responses that either limit or support the pathogen. The infectious bursal disease virus (IBDV) targets primarily the bursa of Fabricius (BF) of chickens, causing severe immunodeficiency. Here, we characterized the cellular proteome changes of the BF caused by IBDV replication in vivo using 2DE followed MALDI-TOF MS identification. Comparative analysis of multiple 2DE gels revealed that the majority of protein expression changes appeared between 24 and 96 h after IBDV infection. MS identified 54 altered cell proteins, 12 of which were notably upregulated by IBDV infection. Meanwhile, the other 42 cellular proteins were considerably suppressed by IBDV infection and are involved in protein degradation, energy metabolism, stress response, host macromolecular biosynthesis, and transport process. The upregulation of β-actin and downregulation of dynamin during IBDV infection were also confirmed by Western blot and immunofluorescence analysis. These altered protein expressions provide a response profile of chicken BF to virulent IBDV infection. Further functional study on these altered proteins may lead to better understanding of pathogenic mechanisms of virulent IBDV infection and to new potential therapeutic targets.     

Dispersal, vaccination and regression of immune defence organs

The thymus in vertebrates and the bursa of Fabricius in birds regress before reproduction, while the immunological information of these organs is maintained as cell memory. Regression at a certain age presupposes that individuals have achieved exposure to a large fraction of parasites in the environment. Here we present a new scenario for regression of immune defence organs, based on optimality reasoning. This scenario links early involution of immune defence organs with (1) effects of exposure to parasites on adaptive immune responses to these parasites, (2) exposure to local parasite communities during natal dispersal and migration as a means of "vaccination" against local parasites, and (3) the function of visits to future breeding sites by juveniles as a means of exposure to local parasites. This scenario provides explanations for why natal dispersal is longer than breeding dispersal, for sex differences in dispersal, and for why the bursa of Fabricius regresses relatively early in life among bird species with delayed start of reproduction.     

Pathology and ultrastructure of Hz-2V infection in the agonadal female corn earworm,Helicoverpa zea

The pathology and ultrastructure of the reproductive tract of Hz-2V-infected female corn earworm moths, Helicoverpa zea, were studied. The identity of malformed reproductive tissues found in virus-infected moths was determined by examining these tissues in moths that were infected with the virus at different life stages. Malformation of reproductive tissues in the progeny of virus-infected female moths was first observed by 3 days post-pupation (dpp), indicating that virus replication had altered the differentiation of these tissues very early on in their development. The ultrastructure of the grossly malformed agonadal reproductive tissues from insects aged 3-10dpp revealed the absence of the cuticular lining found in the oviducts of normal moths, and the proliferation of epithelial cells in these infected oviduct tissues. In addition, large quantities of virus were found aggregated into a large mass in the lumen of the malformed cervix bursa of 10dpp agonadal female pharate adult moths. Prior to eclosion, the virus in the cervix bursa was observed separated into spherical masses, which are thought to exude through the ductus bursa and collect over the vulva, forming a viral "waxy plug" that is likely to play an important role in virus transmission.     

Paradorippe granulata – A crab with external fertilization and a novel type of sperm storage organ challenges prevalent ideas on the evolution of reproduction in Eubrachyura (Crustacea: Brachyura: Dorippidae)

Two fundamentally different sperm storage organs occur in Brachyura. The probably paraphyletic podotremes show intersegmental spermathecae, which are distant from oviducts and coxal gonopores. Hence, fertilization is external. In contrast to this, the seminal receptacles of Eubrachyura are directly connected with the ovaries. Thus, at least initial fertilization is internal. This pattern has been interpreted as an apomorphy of Eubrachyura. To test this hypothesis, we studied the morphology of the reproductive organs of Paradorippe granulata, a representative of the putatively early diverging eubrachyuran lineage Dorippoidea. Applying histology, 3D-reconstructions and micro-computed-tomography we revealed a novel type of sperm storage organ. Female P.granulata lack the characteristic eubrachyuran seminal receptacle. Instead sperm is stored in four cuticle-lined bursae, two on each side of the paired oviducts. The elaborate bulbous male gonopod with several terminal processes is adapted to transferring sperm into the female twin bursae. Since oviducts and twin bursae are not directly connected, spermatozoa and oocytes mix when gametes pass through the sternal vulva. Thus, fertilization in P.granulata is external. Our finding of a eubrachyuran crab that lacks seminal receptacles and exhibits external fertilization calls prevailing concepts on the evolution of sperm storage in Eubrachyura into question.     

关于间质生血干细胞侵入法氏囊上皮及分化成淋巴细胞的研究

在上世纪末Retterer(1885)认为法氏囊的淋巴细胞是法氏囊上皮本身发生的。到本世纪初有人认为从法氏囊间质的“原始成血细胞”侵入上皮分化而成的(Jolly,1915)。直到六十年代初,Ackerman和Knouff(1959),Ackerman(1962),还认为法氏囊髓部的淋巴细胞是上皮细胞发生的,而皮部的淋巴细胞是法氏囊间质细胞及未分化的上皮发生的。用染色体标记等技术证明生血干细胞是法氏囊淋巴细胞的先躯细胞(Moore和Owm1965,1966;Jaffe和Fechhelmer,1966;Le Douarin和Houssaint,1974以及Houssaint等,1976)。生血干细胞在鸡胚发育三天到四天就存在于血液中,只有在法氏囊原基发育到一定阶段才开始侵入(Le Douarin等,1976)。在鸡胚从孵化8天到14天侵入法氏囊原基     

Function of multiple sperm-storage organs in female damselflies (Ischnura senegalensis): difference in amount of ejaculate stored, sperm loss, and priority in fertilization

We studied changes in the number of sperm within two kinds of female sperm-storage organ in the damselfly Ischnura senegalensis (Odonata: Coenagrionidae): the bursa copulatrix and the spermatheca. We counted the number of sperm within each storage organ and tested their viability after a single copulation in female damselflies kept for seven days with and without oviposition. We also counted sperm and tested their viability in females that underwent an interrupted second copulation after the sperm-removal stage, and after subsequent oviposition. Our results showed that the bursa copulatrix and spermatheca have different sperm storage roles. Immediately after copulation, most eggs appear to have been fertilized with bursal sperm, which were positioned near the fertilization point. By seven days after copulation, a greater proportion of spermathecal sperm were used for fertilization, as the number of bursal sperm had decreased. We hypothesize that female damselflies use the spermatheca for long-term storage and the bursa copulatrix for short-term storage: bursal sperm are more likely to be used for fertilization but may have a higher risk of mortality due to sperm removal by a competing male and/or sperm expelling by the female, whereas spermathecal sperm are safer but will be used for fertilization only after their release from the spermatheca.     

关于法氏囊自然退化的研究

实验结果进一步表明法氏囊的发育是受性激素控制的。法氏囊的自然退化是机体内性激素浓度达到一定的水平所致。 在法氏襄的退化过程中起关键作用的细胞成分是FAE细胞,当它们完全地受到性激素的影响时,淋巴细胞分化的微环境遭到破坏,法氏襄发生不可逆性的退化。     

Evidence for a blood-bursa barrier

Summary By use of horseradish peroxidase (Sigma Type II) as a vascular tracer the competence of the bursal microvasculature was evaluated in 15-day-old chicks. Within 5 min circulation time tracer could by identified only within the vascular tree with no leakage into the perivascular or extracellular spaces. After 10 min circulation time tracer was no longer present in the bursal vessels. Since no tracer was ever identified among the parenchymal elements of the organ these data suggest that a blood-organ (blood-bursa) barrier may exist similar to that found in thymus and brain.     

Functional morphology of the copulatory organs of a reed beetle and a shining leaf beetle (Coleoptera: Chrysomelidae: Donaciinae,Criocerinae) using X-ray micro-computed tomography

For more than 100 years it has been known that the sclerotised median lobe of beetles harbours a membranous structure (the "internal sac" or "endophallus") which is everted during copula inside the female genital tract. In order to explore the functional role of this structure and those associated with it, we cryofixed copulating pairs of Donacia semicuprea and Lilioceris lilii and studied the relative position of the elements of the copulatory apparatus of males and females by micro-computer-tomography.We found that the everted endophallus fills the lumen of the bursa copulatrix completely. Our data suggest that in Lilioceris lilii the tip of the sclerotised distal part of the ejaculatory duct, the flagellum, is positioned exactly over the opening of the spermathecal duct inside the bursa copulatrix. The mouth of the bursa copulatrix in Donacia semicuprea is armed with a strong muscle ring, and the whole wall of the bursa is covered externally with a layer of muscle fibres. These morphological differences correspond with differences in mating behaviour: In reed beetles (Donaciinae), females seemingly can control mating to a higher degree than in lily beetles (Lilioceris spp.).