10 results on '"Perner, Jan"'
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2. Haem Biology in Metazoan Parasites – 'The Bright Side of Haem'.
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Perner, Jan, Gasser, Robin B., Oliveira, Pedro L., and Kopáček, Petr
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HEME , *HOST-parasite relationships , *PARASITES , *EMBRYOLOGY , *TICKS , *NEMATODES - Abstract
Traditionally, host haem has been recognized as a cytotoxic molecule that parasites need to eliminate or detoxify in order to survive. However, recent evidence indicates that some lineages of parasites have lost genes that encode enzymes involved specifically in endogenous haem biosynthesis. Such lineages thus need to acquire and utilize haem originating from their host animal, making it an indispensable molecule for their survival and reproduction. In multicellular parasites, host haem needs to be systemically distributed throughout their bodies to meet the haem demands in all cell and tissue types. Host haem also gets deposited in parasite eggs, enabling embryogenesis and reproduction. Clearly, a better understanding of haem biology in multicellular parasites should elucidate organismal adaptations to obligatory blood-feeding. Highlights Ticks and nematodes do not code for haem biosynthetic and degrading enzymes, likely operating independent haem and iron acquisition/distribution networks. The uptake of exogenous haem brings a selective advantage in a haem-rich environment, even in the presence of functional haem biosynthesis. The unavailability of host haem is often manifested during embryogenesis, and the formation of progeny is conditioned by deposits of host haem. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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3. On the haem auxotrophy of the soft tick Ornithodoros moubata.
- Author
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Hatalová, Tereza, Erhart, Jan, Kopáček, Petr, and Perner, Jan
- Abstract
• Transcriptomes of soft ticks encode an incomplete haem biosynthetic pathway. • Haem that Ornithodoros moubata females deposit in their eggs is of host origin and is acquired from host blood haemoglobin. • Depletion of dietary haemoglobin leads to reduced egg-laying capacity of Ornithodoros moubata females. • The genetic coding for porphobilinogen synthase is unique among haem auxotrophs and the enzyme is of unknown function in soft ticks. Genomes of ticks display reductions, to various extents, in genetic coding for enzymes of the haem biosynthetic pathway. Here, we mined available transcriptomes of soft tick species and identified transcripts encoding only half of the enzymes involved in haem biosynthesis. Transcripts identified across most species examined were those coding for porphobilinogen synthase, coproporphyrinogen oxidase, protoporphyrinogen oxidase, and ferrochelatase. Genomic retention of porphobilinogen synthase seems to be soft tick-restricted as no such homologue has been identified in any hard tick species. Bioinformatic mining is thus strongly indicative of the lack of biochemical capacity for de novo haem biosynthesis, suggesting a requirement for dietary haem. In the hard tick Ixodes ricinus , depletion of dietary haem, i.e. serum feeding, leads to oviposition of haem-free eggs, with no apparent embryogenesis and larvae formation. In this work, we show that serum-fed Ornithodoros moubata females, unlike those of I. ricinus , laid haem-containing eggs similarly to blood-fed controls, but only by a small proportion of the serum-fed females. To enhance the effect of dietary haem depletion, O. moubata ticks were serum-fed consecutively as last nymphal instars and females. These females laid eggs with profoundly reduced haem deposits, confirming the host origin of the haem. These data confirm the ability of soft ticks to take up and allocate host haem to their eggs in order to drive reproduction of the ticks. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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4. Inducible glutathione S-transferase (IrGST1) from the tick Ixodes ricinus is a haem-binding protein.
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Perner, Jan, Kotál, Jan, Hatalová, Tereza, Urbanová, Veronika, Bartošová-Sojková, Pavla, Brophy, Peter M., and Kopáček, Petr
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GLUTATHIONE transferase , *CARRIER proteins , *CASTOR bean tick , *NUCLEOTIDE sequence , *ARGASIDAE - Abstract
Blood-feeding parasites are inadvertently exposed to high doses of potentially cytotoxic haem liberated upon host blood digestion. Detoxification of free haem is a special challenge for ticks, which digest haemoglobin intracellularly. Ticks lack a haem catabolic mechanism, mediated by haem oxygenase, and need to dispose of vast majority of acquired haem via its accumulation in haemosomes. The knowledge of individual molecules involved in the maintenance of haem homeostasis in ticks is still rather limited. RNA-seq analyses of the Ixodes ricinus midguts from blood- and serum-fed females identified an abundant transcript of glutathione S-transferase ( gst ) to be substantially up-regulated in the presence of red blood cells in the diet. Here, we have determined the full sequence of this encoding gene, ir-gst1 , and found that it is homologous to the delta-/epsilon-class of GSTs. Phylogenetic analyses across related chelicerates revealed that only one clear Ir GST1 orthologue could be found in each available transcriptome from hard and soft ticks. These orthologues create a well-supported clade clearly separated from other ticks' or mites’ delta-/epsilon-class GSTs and most likely evolved as an adaptation to tick blood-feeding life style. We have confirmed that Ir GST1 expression is induced by dietary haem(oglobin), and not by iron or other components of host blood. Kinetic properties of recombinant Ir GST1 were evaluated by model and natural GST substrates. The enzyme was also shown to bind haemin in vitro as evidenced by inhibition assay, VIS spectrophotometry, gel filtration, and affinity chromatography. In the native state, Ir GST1 forms a dimer which further polymerises upon binding of excessive amount of haemin molecules. Due to susceptibility of ticks to haem as a signalling molecule, we speculate that the expression of Ir GST1 in tick midgut functions as intracellular buffer of labile haem pool to ameliorate its cytotoxic effects upon haemoglobin intracellular hydrolysis. [ABSTRACT FROM AUTHOR]
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- 2018
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5. Multienzyme degradation of host serum albumin in ticks.
- Author
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Sojka, Daniel, Pytelková, Jana, Perner, Jan, Horn, Martin, Konvičková, Jitka, Schrenková, Jana, Mareš, Michael, and Kopáček, Petr
- Abstract
Host blood proteins, represented mainly by hemoglobin and serum albumin, serve as the ultimate source of amino acids needed for de novo protein synthesis during tick development and reproduction. While uptake and processing of hemoglobin by tick gut cells have been studied in detail, molecular mechanisms of host serum albumin degradation remain unknown. In this work, we have used artificial membrane feeding of Ixodes ricinus females on a hemoglobin-free diet in order to characterize the proteolytic machinery involved in albuminolysis. Morphological comparisons of ticks fed on whole blood (BF) and serum (SF) at microscopic and ultrastructural levels showed that albumin and hemoglobin have different trafficking routes in tick gut cells. Analysis in vitro with selective inhibitors demonstrated that albumin is degraded at an acidic pH by a network of cysteine and aspartic peptidases with predominant involvement of cysteine cathepsins having endo- and exopeptidase activities. The cleavage map of albumin and the roles of individual peptidases in albumin degradation were determined. These results indicate that the albuminolytic pathway is controlled by the same proteolytic system that is responsible for hemoglobinolysis. This was further supported by the overall similarity of gut peptidase profiles in SF and BF ticks at the transcriptional and enzymatic activity levels. In conclusion, our work provides evidence that although hemoglobin and albumin are transported differentially during heterophagy they are digested by a common multienzyme proteolytic network. This central digestive system, critical for successful blood feeding in tick females, thus represents a valuable target for novel anti-tick interventions. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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6. Tick iron and heme metabolism – New target for an anti-tick intervention.
- Author
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Hajdusek, Ondrej, Sima, Radek, Perner, Jan, Loosova, Gabriela, Harcubova, Adela, and Kopacek, Petr
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Ticks are blood-feeding parasites and vectors of serious human and animal diseases. Ixodes ricinus is a common tick in Europe, transmitting tick-borne encephalitis, Lyme borreliosis, anaplasmosis, or babesiosis. Immunization of hosts with recombinant tick proteins has, in theory, the potential to interfere with tick feeding and block transmission of pathogens from the tick to the host. However, the efficacy of tick antigens has, to date, not been fully sufficient to achieve this. We have focused on 11 in silico identified genes encoding proteins potentially involved in tick iron and heme metabolism. Quantitative real-time PCR (qRT-PCR) expression profiling was carried out to preferentially target proteins that are up-regulated during the blood meal. RNA interference (RNAi) was then used to score the relative importance of these genes in tick physiology. Finally, we performed vaccination screens to test the suitability of these proteins as vaccine candidates. These newly identified tick antigens have the potential to improve the available anti-tick vaccines. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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7. Functional characterization of the insulin signaling pathway in the hard tick Ixodes ricinus.
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Kozelková, Tereza, Doležel, David, Grunclová, Lenka, Kučera, Matěj, Perner, Jan, and Kopáček, Petr
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Ticks are blood-feeding arachnids transmitting a variety of pathogens to humans and animals. A unique trait in tick physiology is their ability to engorge and digest large amounts of host blood, ensuring their high reproductive potential. Activation of the blood digestive machinery in the tick gut, as well as processes controlling maturation of ovaries, are triggered upon blood meal uptake by still largely unknown mechanisms. Sensing of the nutritional status in metazoan organisms is facilitated by the evolutionarily conserved Insulin Signaling Pathway (ISP) and the interlinked Target of Rapamycin (TOR) pathway. Recently, we have identified three components of these pathways in the hard tick Ixodes ricinus midgut transcriptome, namely a putative insulin receptor (InR), and the downstream intracellular serine/threonine kinases AKT and TOR. In this study, we primarily focus on the molecular and functional characterization of the I. ricinus insulin receptor (Ir InR), the first InR characterized in Chelicerates. A phylogenetic analysis across the major Arthropod lineages demonstrated that ticks possess only one gene encoding an InR-related molecule. Tissue expression profiling by quantitative PCR in semi-engorged I. ricinus females revealed that the Ir InR, as well as AKT (Ir AKT) and TOR (Ir TOR) are expressed in various organs, with the highest expression being detected in ovaries. We have further evaluated the impact of RNAi-mediated knock-down (KD) of Ir InR, Ir AKT, and Ir TOR on tick blood-feeding and reproductive capacity. Weights of engorged Ir InR KD females and laid egg clutches were reduced compared to the control group, and these quantitative parameters clearly correlated with the efficiency of RNAi-KD achieved in individual ticks. The most striking phenotype was observed for Ir AKT KD that impaired tick feeding and completely aborted egg production. A recombinant extracellular fragment of the Ir InR α-subunit was used to produce antibodies in experimental rabbits to assess its potential as a protective antigen against tick feeding and reproduction. Our data clearly indicate the functionality of the ISP in ticks and demonstrate the need for further investigation of specific roles played by the endogenous insulin-like peptides in tick physiological processes. [ABSTRACT FROM AUTHOR]
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- 2021
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8. From the fat body to the hemolymph: Profiling tick immune and storage proteins through transcriptomics and proteomics.
- Author
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Urbanová, Veronika, Lu, Stephen, Kalinová, Eliška, Martins, Larissa, Kozelková, Tereza, Dyčka, Filip, Ribeiro, José M., Hajdušek, Ondřej, Perner, Jan, and Kopáček, Petr
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FAT , *TRANSCRIPTOMES , *TICK infestations , *HEMOLYMPH , *PROTEOMICS , *TICKS , *PLANT defenses - Abstract
Ticks are blood-feeding arachnids that are known to transmit various pathogenic microorganisms to their hosts. During blood feeding, ticks activate their metabolism and immune system to efficiently utilise nutrients from the host's blood and complete the feeding process. In contrast to insects, in which the fat body is known to be a central organ that controls essential metabolic processes and immune defense mechanisms, the function of the fat body in tick physiology is still relatively unexplored. To fill this gap, we sought to uncover the repertoire of genes expressed in the fat body associated with trachea (FB/Tr) by analyzing the transcriptome of individual, partially fed (previtellogenic) Ixodes ricinus females. The resulting catalog of individual mRNA sequences reveals a broad repertoire of transcripts encoding proteins involved in nutrient storage and distribution, as well as components of the tick immune system. To gain a detailed insight into the secretory products of FB/Tr specifically involved in inter-tissue transport and humoral immunity, the transcriptomic data were complemented with the proteome of soluble proteins in the hemolymph of partially fed female ticks. Among these proteins, the hemolipoglyco-carrier proteins were predominant. When comparing immune peptides and proteins from the fat body with those produced by hemocytes, we found that the fat body serves as a unique producer of certain immune components. Finally, time-resolved transcriptional regulation of selected immune transcripts from the FB/Tr was examined in response to experimental challenges with model microbes and analyzed by RT-qPCR. Overall, our data show that the fat body of ticks, similar to insects, is an important metabolic tissue that also plays a remarkable role in immune defense against invading microbes. These findings improve our understanding of tick biology and its impact on the transmission of tick-borne pathogens. [Display omitted] • The first transcriptomic study of the fat body/trachea (FB/Tr) complex of ticks was performed. • Transcriptomic data were complemented by proteomic analysis of the hemolymph. • Hemolipoglyco-carrier proteins (HeLp) produced in the FB/Tr complex were a predominant fraction of tick plasma. • Expression of FB/Tr transcripts encoding immune genes responded to microbial challenge. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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9. Dual SIFamide receptors in Ixodes salivary glands.
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Guerrib, Fetta, Ning, Caina, Mateos-Hernandéz, Lourdes, Rakotobe, Sabine, Park, Yoonseong, Hajdusek, Ondrej, Perner, Jan, Vancová, Marie, Valdés, James J., and Šimo, Ladislav
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SALIVARY glands , *IXODES , *IXODES scapularis , *CASTOR bean tick , *EPITHELIAL cells , *PEPTIDES - Abstract
Salivary glands are vital to tick feeding success and also play a crucial role in tick-borne pathogen transmission. In previous studies of Ixodes scapularis salivary glands, we demonstrated that saliva-producing type II and III acini are innervated by neuropeptidergic axons which release different classes of neuropeptides via their terminals (Šimo et al., 2009b, 2013). Among these, the neuropeptide SIFamide—along with its cognate receptor—were postulated to control the basally located acinar valve via basal epithelial and myoepithelial cells (Vancová et al., 2019). Here, we functionally characterized a second SIFamide receptor (SIFa_R2) from the I. scapularis genome and proved that it senses a low nanomolar level of its corresponding ligand. Insect SIFamide paralogs, SMYamides, also activated the receptor but less effectively compared to SIFamide. Bioinformatic and molecular dynamic analyses suggested that I. scapularis SIFamide receptors are class A GPCRs where the peptide amidated carboxy-terminus is oriented within the receptor binding cavity. The receptor was found to be expressed in Ixodes ricinus salivary glands, synganglia, midguts, trachea, and ovaries, but not in Malpighian tubules. Investigation of the temporal expression patterns suggests that the receptor transcript is highly expressed in unfed I. ricinus female salivary glands and then decreases during feeding. In synganglia, a significant transcript increase was detected in replete ticks. In salivary gland acini, an antibody targeting the SIFa_R2 recognized basal epithelial cells, myoepithelial cells, and basal granular cells in close proximity to the SIFamide-releasing axon terminals. Immunoreactivity was also detected in specific neurons distributed throughout various I. ricinus synganglion locations. The current findings, alongside previous reports from our group, indicate that the neuropeptide SIFamide acts via two different receptors that regulate distinct or common cell types in the basal region of type II and III acini in I. ricinus salivary glands. Our study investigates the peptidergic regulation of the I. ricinus salivary gland in detail, emphasizing the complexity of this system. [Display omitted] • Ixodes scapularis SIFamide receptor 2 (SIFa_R2) displays high affinity for SIFamide as well as to its insect paralogs. • I. scapularis SIFa_R1 displays exclusive affinity for SIFamide and not to its insect paralogs. • SIFa_R2 is expressed in I. ricinus salivary glands, synganglion, midguts, trachea, and ovaries. • SIFa_R2 is localized in distinct basal cell types in both type II and III salivary gland acini. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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10. Multiple legumain isoenzymes in ticks.
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Hartmann, David, Šíma, Radek, Konvičková, Jitka, Perner, Jan, Kopáček, Petr, and Sojka, Daniel
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ISOENZYMES , *TICKS , *LYME disease , *IXODES scapularis , *MESSENGER RNA - Abstract
By searching nucleotide databases for the North American Lyme disease vector, Ixodes scapularis , we have complemented the previously characterized European Ixodes ricinus legumain IrAE1 with a full set of nine analogous genes ( isae1-9 ). Six of these were PCR confirmed as genes present in all tick genomes tested. The absolute mRNA copy number examined by quantitative (q)PCR enabled expression profiling and an absolute comparison of mRNA levels for individual I. scapularis (Is)AEs in tick tissues. Four IsAEs (1, 2, 4, 9) were expressed solely in the gut and thus are proposed to be involved in host blood digestion. Expression qPCR profiling over developmental stages confirmed IsAE1, the direct analogue of previously characterized I. ricinus IrAE1, as the principle legumain transcript in partially engorged females, and demonstrated its strong regulation by on-host feeding in larvae, nymphs and females. In contrast, IsAE2 was the predominant gut legumain in unfed nymphs, unfed females and males. In-silico, IsAE1 and IsAE2 protein three-dimensional structural models displayed minimal differences in overall proenzyme structures, even in comparison with recently resolved crystal structures of mammalian prolegumain. Three functional studies were performed in I. ricinus with IsAE1/IsAE2 analogues: double IrAE1/IrAE2 RNA interference silencing, feeding of ticks on IrAE1+IrAE2 immunized hosts and in vitro membrane tick feeding on blood containing a legumain-specific inhibitor. The latter experiment led to reduced weights of fully engorged ticks and limited oviposition, and indicated the potential of legumain inhibitors for novel anti-tick interventions. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
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