13 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. Sialome diversity of ticks revealed by RNAseq of single tick salivary glands.
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Perner, Jan, Kropáčková, Sára, Kopáček, Petr, and Ribeiro, José M. C.
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TICKS as carriers of disease , *SALIVARY glands , *PARASITES , *IMMUNE response , *RELAPSING fever - Abstract
Ticks salivate while feeding on their hosts. Saliva helps blood feeding through host anti-hemostatic and immunomodulatory components. Previous transcriptomic and proteomic studies revealed the complexity of tick saliva, comprising hundreds of polypeptides grouped in several multi-genic families such as lipocalins, Kunitz-domain containing peptides, metalloproteases, basic tail secreted proteins, and several other families uniquely found in ticks. These studies also revealed that the composition of saliva changes with time; expression of transcripts from the same family wax and wane as a function of feeding time. Here, we examined whether host immune factors could influence sialome switching by comparing sialomes of ticks fed naturally on a rabbit, to ticks artificially fed on defibrinated blood depleted of immune components. Previous studies were based on transcriptomes derived from pools of several individuals. To get an insight into the uniqueness of tick sialomes, we performed transcriptomic analyses of single salivary glands dissected from individual adult female I. ricinus ticks. Multivariate analysis identified 1,279 contigs differentially expressed as a function of time and/or feeding mode. Cluster analysis of these contigs revealed nine clusters of differentially expressed genes, four of which appeared consistently across several replicates, but five clusters were idiosyncratic, pointing to the uniqueness of sialomes in individual ticks. The disclosure of tick quantum sialomes reveals the unique salivary composition produced by individual ticks as they switch their sialomes throughout the blood meal, a possible mechanism of immune evasion. [ABSTRACT FROM AUTHOR]
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- 2018
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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]
- Published
- 2018
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5. Acquisition of exogenous haem is essential for tick reproduction.
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Perner, Jan, Sobotka, Roman, Sima, Radek, Konvickova, Jitka, Sojka, Daniel, de Oliveira, Pedro Lagerblad, Hajdusek, Ondrej, and Kopacek, Petr
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ABORTION , *EUKARYOTIC cells , *EUKARYOTES , *PROTISTA , *NUCLEAR DNA - Abstract
Haem and iron homeostasis in most eukaryotic cells is based on a balanced flux between haem biosynthesis and haem oxygenase-mediated degradation. Unlike most eukaryotes, ticks possess an incomplete haem biosynthetic pathway and, together with other (non-haematophagous) mites, lack a gene encoding haem oxygenase. We demonstrated, by membrane feeding, that ticks do not acquire bioavailable iron from haemoglobin-derived haem. However, ticks require dietary haemoglobin as an exogenous source of haem since, feeding with haemoglobin-depleted serum led to aborted embryogenesis. Supplementation of serum with haemoglobin fully restored egg fertility. Surprisingly, haemoglobin could be completely substituted by serum proteins for the provision of amino-acids in vitellogenesis. Acquired haem is distributed by haemolymph carrier protein(s) and sequestered by vitellins in the developing oocytes. This work extends, substantially, current knowledge of haem auxotrophy in ticks and underscores the importance of haem and iron metabolism as rational targets for anti-tick interventions. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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6. Variation of bacterial community assembly over developmental stages and midgut of Dermanyssus gallinae.
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Wu-Chuang, Alejandra, Hartmann, David, Maitre, Apolline, Mateos-Hernández, Lourdes, Frantová, Helena, Urbanová, Veronika, Obregon, Dasiel, Cabezas-Cruz, Alejandro, and Perner, Jan
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GALLIFORMES , *LIFE cycles (Biology) , *BACTERIAL diversity , *BACTERIAL communities , *MICROBIAL communities , *MITES - Abstract
Bacterial microbiota play an important role in the fitness of arthropods, but the bacterial microflora in the parasitic mite Dermanyssus gallinae is only partially explored; there are gaps in our understanding of the microbiota localization and in our knowledge of microbial community assembly. In this work, we have visualized, quantified the abundance, and determined the diversity of bacterial occupancy, not only across developmental stages of D. gallinae, but also in the midgut of micro-dissected female D. gallinae mites. We explored community assembly and the presence of keystone taxa, as well as predicted metabolic functions in the microbiome of the mite. The diversity of the microbiota and the complexity of co-occurrence networks decreased with the progression of the life cycle. However, several bacterial taxa were present in all samples examined, indicating a core symbiotic consortium of bacteria. The relatively higher bacterial abundance in adult females, specifically in their midguts, implicates a function linked to the biology of D. gallinae mites. If such an association proves to be important, the bacterial microflora qualifies itself as an acaricidal or vaccine target against this troublesome pest. [ABSTRACT FROM AUTHOR]
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- 2023
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7. RNA-seq analyses of the midgut from blood- and serum-fed Ixodes ricinus ticks.
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Perner, Jan, Provazník, Jan, Schrenková, Jana, Urbanová, Veronika, Ribeiro, José M. C., and Kopáček, Petr
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Adult females of the genus Ixodes imbibe blood meals exceeding about 100 times their own weight within 7-9 days. During this period, ticks internalise components of host blood by endocytic digest cells that line the tick midgut epithelium. Using RNA-seq, we aimed to characterise the midgut transcriptome composition in adult Ixodes ricinus females during early and late phase of engorgement. To address specific adaptations to the haemoglobin-rich diet, we compared the midgut transcriptomes of genetically homogenous female siblings fed either bovine blood or haemoglobin-depleted serum. We noted that tick gut transcriptomes are subject to substantial temporal-dependent expression changes between day 3 and day 8 of feeding. In contrast, the number of transcripts significantly affected by the presence or absence of host red blood cells was low. Transcripts relevant to the processes associated with blood-meal digestion were analysed and involvement of selected encoded proteins in the tick midgut physiology discussed. A total of 7215 novel sequences from I. ricinus were deposited in public databases as an additional outcome of this study. Our results broaden the current knowledge of tick digestive system and may lead to the discovery of potential molecular targets for efficient tick control. [ABSTRACT FROM AUTHOR]
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- 2016
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8. Vector Biology: Tyrosine Degradation Protects Blood Feeders from Death via La Grande Bouffe.
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Kopáček, Petr and Perner, Jan
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GENETIC vectors , *CONENOSES , *DISEASE vectors , *TYROSINE , *MOSQUITO vectors - Abstract
Summary Blood-feeding arthropods digest vast amounts of host-blood nutrients. A new study suggests that tyrosine degradation is essential for the survival of blood-fed kissing bugs, mosquitoes, and ticks. This finding presents a promising target for the control of these disease vectors. [ABSTRACT FROM AUTHOR]
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- 2016
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9. Blood-feeding adaptations and virome assessment of the poultry red mite Dermanyssus gallinae guided by RNA-seq.
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Ribeiro, José M., Hartmann, David, Bartošová-Sojková, Pavla, Debat, Humberto, Moos, Martin, Šimek, Petr, Fara, Jiří, Palus, Martin, Kučera, Matěj, Hajdušek, Ondřej, Sojka, Daniel, Kopáček, Petr, and Perner, Jan
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GALLIFORMES , *FERRITIN , *CATHEPSIN B , *CYSTEINE proteinases , *MITES , *VITELLOGENINS - Abstract
Dermanyssus gallinae is a blood-feeding mite that parasitises wild birds and farmed poultry. Its remarkably swift processing of blood, together with the capacity to blood-feed during most developmental stages, makes this mite a highly debilitating pest. To identify specific adaptations to digestion of a haemoglobin-rich diet, we constructed and compared transcriptomes from starved and blood-fed stages of the parasite and identified midgut-enriched transcripts. We noted that midgut transcripts encoding cysteine proteases were upregulated with a blood meal. Mapping the full proteolytic apparatus, we noted a reduction in the suite of cysteine proteases, missing homologues for Cathepsin B and C. We have further identified and phylogenetically analysed three distinct transcripts encoding vitellogenins that facilitate the reproductive capacity of the mites. We also fully mapped transcripts for haem biosynthesis and the ferritin-based system of iron storage and inter-tissue trafficking. Additionally, we identified transcripts encoding proteins implicated in immune signalling (Toll and IMD pathways) and activity (defensins and thioester-containing proteins), RNAi, and ion channelling (with targets for commercial acaricides such as Fluralaner, Fipronil, and Ivermectin). Viral sequences were filtered from the Illumina reads and we described, in part, the RNA-virome of D. gallinae with identification of a novel virus, Red mite quaranjavirus 1. Genes associated with the consumption of blood and iron storage are identified in the poultry red mite, along with presentation of a partial virome. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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10. From the fat body to the hemolymph: Profiling tick immune and storage proteins through transcriptomics and proteomics.
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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]
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- 2024
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11. 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]
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- 2018
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12. 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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13. Tick-borne encephalitis virus infects human brain microvascular endothelial cells without compromising blood-brain barrier integrity.
- Author
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Palus, Martin, Vancova, Marie, Sirmarova, Jana, Elsterova, Jana, Perner, Jan, and Ruzek, Daniel
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TICK-borne encephalitis viruses , *BLOOD-brain barrier , *CELL adhesion molecules , *TRANSCYTOSIS , *BRAIN physiology - Abstract
Alteration of the blood-brain barrier (BBB) is a hallmark of tick-borne encephalitis (TBE), a life-threating human viral neuroinfection. However, the mechanism of BBB breakdown during TBE, as well as TBE virus (TBEV) entry into the brain is unclear. Here, primary human microvascular endothelial cells (HBMECs) were infected with TBEV to study interactions with the BBB. Although the number of infected cells was relatively low in culture (<5%), the infection was persistent with high TBEV yields (>10 6 pfu/ml). Infection did not induce any significant changes in the expression of key tight junction proteins or upregulate the expression of cell adhesion molecules, and did not alter the highly organized intercellular junctions between HBMECs. In an in vitro BBB model, the virus crossed the BBB via a transcellular pathway without compromising the integrity of the cell monolayer. The results indicate that HBMECs may support TBEV entry into the brain without altering BBB integrity. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
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