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1.
Application of RNA interference in tick salivary gland research. 总被引:3,自引:0,他引:3
Vijay G Ramakrishnan Majd N Aljamali John R Sauer Richard C Essenberg 《Journal of biomolecular techniques》2005,16(4):297-305
Ticks are obligate ectoparasites that feed on a variety of hosts including mammals, birds and reptiles. Prolonged attachment on the host and an ability to transmit a wide variety of pathogens are the special features of tick feeding. Salivary glands are the major route for secretion of excess fluid, several proteins, and factors that counteract the host immune response and hence play a significant role in the success of tick feeding. RNA interference (RNAi) enables scientists to silence genes encoding proteins in an absolutely sequence specific manner at the mRNA level. This technique has already been successfully employed in analyzing roles of proteins of important functions or in assigning roles to several proteins of unknown functions in a variety of animals. In this review, we outline the process of RNAi and the applicability of RNAi in tick salivary gland research. 相似文献
2.
J. R. Sauer J. L. McSwain J. S. Tucker K. S. Shelby J. P. Williams R. C. Essenberg 《Experimental & applied acarology》1989,7(1):81-94
Tick salivary glands are controlled by nerves, dopamine being a neurotransmitter at the neuroeffector junction. Dopamine and cyclic AMP (cAMP) stimulate fluid secretion by isolated salivary glands. Dopamine activates an adenylate cyclase to increase intracellular cAMP within the female salivary glands. Phosphoproteins whose levels of phosphate are affected by cAMP-dependent protein kinase have been identified in subcellular fractions. Protein(s) phosphorylated by cAMP appears to activate protein phosphatase in the salivary glands.Another phosphorylation pathway appears to act through protein kinase C because of an ability of phorbol esters (known activators of protein kinase C) to stimulate the phosphorylation of proteins, and an ability of a peptide factor in tick brain to metabolize salivary-gland phosphoinositides, an event that often precedes activation of protein kinase C. Because cAMP modulates brain-factor-stimulated formation of inositol phosphates (products of phosphoinositide breakdown) an interrelationship between the two pathways seems likely.Evidence of regulatory processes, including protein phosphorylation'dephosphorylation reactions, will provide a basis for helping asses the physiological significance of secretory products and the role of the salivary glands in disease transmission. 相似文献
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In the infested skin of tick-resistant guinea pigs, indirect immunofluorescence techniques have revealed that antigens from the ticks' salivary glands are associated with discrete dendritic cells in the epidermis. Evidence is presented to support the suggestion that these antigen-retaining cells are Langerhans cells. 相似文献
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Thirty-five strains of the Lyme disease spirochete Borrelia burgdorferi sensu lato (B. burgdorferi s. l.) were isolated from the blacklegged tick vector Ixodes scapularis in South Carolina, Georgia, Florida, and Rhode Island. They were characterized by PCR-restriction fragment length polymorphism (RFLP) analysis of rrf (5S)-rrl (23S) intergenic spacer amplicons. PCR-RFLP analysis indicated that the strains represented at least 3 genospecies (including a possible novel genospecies) and 4 different restriction patterns. Thirty strains belonged to the genospecies B. burgdorferi sensu stricto (B. burgdorferi s. s.), 4 southern strains were identified as B. bissettii, and strain SCCH-5 from South Carolina exhibited MseI and DraI restriction patterns different from those of previously reported genospecies. Complete sequences of rrf-rrl intergenic spacers from 14 southeastern and northeastern strains were determined and the phylogenetic relationships of these strains were compared. The 14 strains clustered into 3 separate lineages on the basis of sequence analysis. These results were confirmed by phylogenetic analysis based on 16S rDNA sequence analysis. 相似文献
5.
Severinová J Salát J Krocová Z Reznícková J Demová H Horká H Kopecký J 《Folia microbiologica》2005,50(5):457-463
The impact of Ixodes ricinus salivary gland extract (SGE) on inflammatory changes in the skin and draining lymph nodes of mice, elicited by the infection with the important human pathogen, B. afzelii, was determined using flow cytometry. SGE injected together with spirochetes reduced the numbers of leukocytes and gammadelta-T lymphocytes in infected epidermis at early time-points post infection. In draining lymph nodes, the anti-inflammatory effect of SGE was manifested by the decrease of total cell count compared with that in mice treated with inactivated SGE. Changes in subpopulations of immunocompetent cells apparently reflected the effect of SGE on the proliferation of spirochetes in the host. The significance of tick saliva anti-inflammatory effect for saliva activated transmission of B. afzelii is shown. 相似文献
6.
Putative new expression of genes in ixodid tick salivary gland development during feeding. 总被引:2,自引:0,他引:2
J F Oaks J L McSwain J A Bantle R C Essenberg J R Sauer 《The Journal of parasitology》1991,77(3):378-383
Poly(A+) mRNA-enriched fractions from salivary glands of partially fed Amblyomma americanum female ticks were translated in vitro with a rabbit reticulocyte translation system. Translated proteins were labeled with [35S]methionine, separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and identified by autoradiography. Thirty major identifiable polypeptides with molecular weights ranging from 14 to 136 kDa were synthesized from mRNA isolated from salivary glands of ticks weighing less than 100 mg. Polypeptides that comigrated at the same molecular weight were translated by mRNA from ticks at a more advanced stage of feeding (more than 300 mg) as were 8 others with molecular weights of 31, 71, 91, 106, 113, 118, and 128 kDa. Results demonstrated that differential gene expression may be stimulated in the developing salivary glands as the tick feeds. 相似文献
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Pooled tissue samples are frequently used in biochemical studies involving parasites in order to ensure that there is sufficient material for experimentation. A pooled sample is considered to represent the overall phenotypic characteristics of the investigated population. However, this will not be the case if there is a significant degree of molecular polymorphism among individuals in the sampled population. Here we demonstrate marked differences in the protein profile of salivary glands among individuals from three species of ixodid tick (Rhipicephalus appendiculatus, Amblyomma variegatum, Ixodes ricinus), and show that pooling the tissue of several individuals masks substantial qualitative differences among the individuals. Our observations indicate that much greater caution is needed in general when using pooled samples if the molecular diversity within the population is not clearly defined. 相似文献
9.
S J Brown S Z Shapiro P W Askenase 《Journal of immunology (Baltimore, Md. : 1950)》1984,133(6):3319-3325
Guinea pigs immunized by subcutaneous injection of an emulsion of incomplete Freund's adjuvant (IFA) containing tick salivary gland extract antigens (SGA) from partially fed female ticks expressed a significant level of tick rejection when challenged 17 days later. This level of tick rejection was similar to animals actively sensitized by tick feeding and challenged at the same time. SGA emulsified with complete Freund's adjuvant (CFA) or administered with saline was ineffective. However, ticks that fed on animals immunized with SGA+IFA or SGA+CFA expressed significant reductions in engorgement weight. SGA was active when prepared with or without protease inhibitors. The minimum effective immunizing dose of SGA was between 100 and 280 micrograms per animal. Extracts made from salivary gland-derived cement material (CA) from partially fed female ticks administered at 50 micrograms in IFA induced levels of tick rejection comparable to animals immunized with 280 micrograms of SGA+IFA. Sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS/PAGE) of 35S- and 125I-radiolabeled SGA and CA extracts immunoprecipitated by guinea pig anti-tick serum that transferred immune resistance demonstrated a unique protein of 20,000 m.w. Serum from animals immunized with SGA+IFA (successful immunization) recognized this same protein, whereas serum from animals immunized with SGA+CFA (unsuccessful immunization) did not. The results of this study suggest that a 20,000 m.w. protein derived from the tick salivary gland may be responsible for the induction and perhaps elicitation of host immune resistance responses to Amblyomma americanum ticks. 相似文献
10.
W. Burgdorfer 《The Yale journal of biology and medicine》1984,57(4):515-520
The various hypotheses concerning the etiologic agent of erythema chronicum migrans of Europe and of Lyme disease in the United States are reviewed, and an account of events that led to the discovery of the causative spirochetal agent in Ixodes dammini is presented. Spirochetes morphologically and antigenically similar, if not identical to, the organism detected in I. dammini were also found for the first time in Ixodes pacificus and Ixodes ricinus, the vectors hitherto incriminated, respectively, in western United States and Europe. In most infected ticks, spirochetal development was found to be limited to the midgut. Ticks with generalized infections were shown to transmit spirochetes via eggs, but infections decreased in intensity and became restricted to the central ganglion as filial ticks developed to adults. Although the mechanisms of transmission to a host are still under investigation, the spirochetes may be transmitted by saliva of ticks with generalized infectious and possibly also by regurgitation of infected gut contents, or even by means of infected fecal material. 相似文献
11.
Dermacentor variabilis: regulation of fibroblast migration by tick salivary gland extract and saliva
Kramer C Nahmias Z Norman DD Mulvihill TA Coons LB Cole JA 《Experimental parasitology》2008,119(3):391-397
We examined the effects of tick SGx and saliva on basal- and platelet-derived growth factor (PDGF)-stimulated cell migration and extracellular signal-regulated kinase (ERK) signaling in fibroblasts. Repair of injured monolayers was delayed by SGx pretreatment and was not associated with reductions in cell number. In migration assays, SGx suppressed both basal- and PDGF-stimulated fibroblast movement. Furthermore, SGx and saliva reduced PDGF-stimulated ERK activity. Thus, the delayed repair of monolayer injuries resulted from SGx inhibiting fibroblast migratory responses to chemotactic signals. SGx also suppressed injury- and growth factor-induced ERK activation in renal epithelial OK cells. Our data suggest that maintenance of the tick feeding lesion results, in part, from suppressing ERK signaling and fibroblast migration, events playing integral roles in the wound healing response. The effects of SGx on cells not involved in wound healing suggest that a constituent(s) in tick saliva has global effects on the ERK signaling pathway. 相似文献
12.
Under normal circumstances, salivary glands of female ixodid ticks begin degenerating within hours of completing the blood meal. We have monitored cytological, functional and biochemical changes in the tissue which are diagnostic of the degenerative process. Although ultimately degeneration also befalls salivary glands of partially fed ticks removed prematurely from the host, the process is considerably delayed. When we transplanted salivary glands from partially fed ticks into the haemocoels of replete specimens, autolysis was induced in the donor tissue, whereas such was not the case when similar glands were transplanted to the haemocoels of other partially fed ticks. We thus suggest that a humoral factor is involved in postprandial resorption of the salivary glands. Succinate dehydrogenase activity decreases, and acid phosphatase activity increases in the salivary glands as a function of time post-engorgement. However, these enzyme assays are not sensitive enough to detect the earliest stages of autolysis. 相似文献
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W. Reuben Kaufman B. Duff Sloley Roger J. Tatchell Geoff L. Zbitnew Thomas J. Diefenbach Jeffrey I. Goldberg 《Experimental & applied acarology》1999,23(3):251-265
Dopamine (DA) content of the salivary glands in partially fed female and fed male ticks, Amblyomma hebraeum Koch (Acari: Ixodidae), was measured by high-performance liquid chromatography with electrochemical detection or by a radioenzymatic assay for catecholamines following experimental treatment. Some glands were held in vitro for up to 3 days. Other preparations (backless explants) allowed one side to be surgically denervated, the contralateral side serving as control. Normal ticks were sampled for up to 4 days post-removal from the host (rabbits). In the backless explants, there was little if any difference in DA content between denervated and control sides, even after 4 days in vitro, indicating that unilateral denervation did not eliminate the major salivary gland pool of DA. High doses of reserpine (333 g per g body weight) and 6-hydroxydopamine (1000 g per g body weight) did not significantly reduce the DA content of the salivary gland, also suggesting that only a minor component of the DA pool is within axons innervating the salivary gland. A dispersed population of cells rich in tyrosine hydroxylase immunoreactivity (an enzyme marker for catecholamine-synthesizing cells) was found in close association with the granular acini. This further suggests that the major DA pool in the salivary gland may be in cells other than the dopaminergic nerves arising from the central nervous system. © Rapid Science Ltd. 1998 相似文献
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Changes in antigenic reactivity of Borrelia burgdorferi, the Lyme disease spirochete, during persistent infection in mice 总被引:7,自引:0,他引:7
T G Schwan R H Karstens M E Schrumpf W J Simpson 《Canadian journal of microbiology》1991,37(6):450-454
Adult laboratory mice, Mus musculus, were shown to be suitable experimental animals for studying infectivity, persistent infection, and in vivo antigenic changes of Borrelia burgdorferi. Sixteen mice were inoculated intraperitoneally with a low-passage culture of an uncloned strain of B. burgdorferi and 16 months later spirochetes were reisolated from the urinary bladder of 15 (94%) of the mice. Spirochetes recovered from the urinary bladder of one persistently infected mouse were tested for infectivity and found to be infectious when passaged into four laboratory mice. Western blot analysis of immune serum from each of the persistently infected mice demonstrated that spirochetes used to infect the mice reacted differently when compared with the spirochetes subsequently reisolated from the mice, demonstrating for the first time that changes in antigenic reactivity had occurred in the spirochete populations during persistent infection. 相似文献
18.
Over 80% of reported cases of Lyme disease in the United States occur in coastal regions of northeastern and mid-Atlantic states. The genetic structure of the Lyme disease spirochete (Borrelia burgdorferi) and its main tick vector (Ixodes scapularis) was studied concurrently and comparatively by sampling natural populations of I. scapularis ticks along the East Coast from 1996 to 1998. Borrelia is genetically highly diverse at the outer surface protein ospC. Since Borrelia is highly clonal, the ospC alleles can be used to define clones. A newly designed reverse line blotting (RLB) assay shows that up to 10 Borrelia clones can infect a single tick. The clone frequencies in Borrelia populations are the same across the Northeast. On the other hand, I. scapularis populations show strong regional divergence (among northeastern, mid-Atlantic, and southern states) as well as local differentiation. The high genetic diversity within Borrelia populations and the disparity in the genetic structure between Borrelia and its tick vector are likely consequences of strong balancing selection on local Borrelia clones. Demographically, both Borrelia and I. scapularis populations in the Northeast show the characteristics of a species that has recently expanded from a population bottleneck. Major geological and ecological events, such as the last glacial maximum (18,000 years ago) and the modern-day expansion of tick habitats, are likely causes of the observed "founder effects" for the two organisms in the Northeast. We therefore conclude that the genetic structure of B. burgdorferi has been intimately shaped by the natural history of its main vector, the northern lineage of I. scapularis ticks. 相似文献
19.
Previous work has indicated that both Borrelia burgdorferi and the process of tick feeding (saliva) modulate the host immune response. Molecules have been identified in tick saliva that effect T cell proliferation by binding to specific cytokines, thereby promoting a Th2 cytokine response that does not afford protection against tick-transmitted B. burgdorferi in mice. Moreover, reconstitution of a Th1-biased T cell response prior to spirochete challenge effectively neutralizes tick modulation of host immunity and affords protection against tick transmission of spirochetes. The current studies were undertaken to determine the effect of neutralizing specific Th2 cytokines prior to tick feeding and subsequent transmission of B. burgdorferi. The results indicate that suppression of both IL-4 and IL-5 prior to the feeding of B. burgdorferi-infected ticks significantly decreased spirochete load in target organs such as joint, bladder, heart, and skin of the Lyme disease-susceptible host. 相似文献
20.
Sumita Y Liu Y Khalili S Maria OM Xia D Key S Cotrim AP Mezey E Tran SD 《The international journal of biochemistry & cell biology》2011,43(1):80-87
Treatment for most patients with head and neck cancers includes ionizing radiation. A consequence of this treatment is irreversible damage to salivary glands (SGs), which is accompanied by a loss of fluid-secreting acinar-cells and a considerable decrease of saliva output. While there are currently no adequate conventional treatments for this condition, cell-based therapies are receiving increasing attention to regenerate SGs. In this study, we investigated whether bone marrow-derived cells (BMDCs) can differentiate into salivary epithelial cells and restore SG function in head and neck irradiated mice. BMDCs from male mice were transplanted into the tail-vein of 18Gy-irradiated female mice. Salivary output was increased in mice that received BMDCs transplantation at week 8 and 24 post-irradiation. At 24 weeks after irradiation (IR), harvested SGs (submandibular and parotid glands) of BMDC-treated mice had greater weights than those of non-treated mice. Histological analysis shows that SGs of treated mice demonstrated an increased level of tissue regenerative activity such as blood vessel formation and cell proliferation, while apoptotic activity was increased in non-transplanted mice. The expression of stem cell markers (Sca-1 or c-kit) was detected in BMDC-treated SGs. Finally, we detected an increased ratio of acinar-cell area and approximately 9% of Y-chromosome-positive (donor-derived) salivary epithelial cells in BMDC-treated mice. We propose here that cell therapy using BMDCs can rescue the functional damage of irradiated SGs by direct differentiation of donor BMDCs into salivary epithelial cells. 相似文献