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.

Institute of Entomology Biology Centre of the Czech Academy of Sciences Branišovská 31 370 05 České Budějovice Czech Republic

Institute of Parasitology Biology Centre of the Czech Academy of Sciences Branišovská 31 370 05 České Budějovice Czech Republic

Section of Vector Biology Laboratory of Malaria and Vector Research National Institute of Allergy and Infectious Diseases National Institutes of Health Bethesda Maryland United States of America

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Mans B. J. & Neitz A. W. Adaptation of ticks to a blood-feeding environment: evolution from a functional perspective. Insect biochemistry and molecular biology 34, 1–17 (2004). PubMed

de la Fuente J., Estrada-Pena A., Venzal J. M., Kocan K. M. & Sonenshine D. E. Overview: Ticks as vectors of pathogens that cause disease in humans and animals. Frontiers in bioscience: a journal and virtual library 13, 6938–6946 (2008). PubMed

Franta Z. et al. Dynamics of digestive proteolytic system during blood feeding of the hard tick PubMed DOI PMC

Sonenshine D. E. & Roe R. M.

Brown M. R., Sieglaff D. H. & Rees H. H. Gonadal ecdysteroidogenesis in arthropoda: occurrence and regulation. Annu Rev Entomol 54, 105–125, doi: 10.1146/annurev.ento.53.103106.093334 (2009). PubMed DOI PMC

Schwarz A. et al. A systems level analysis reveals transcriptomic and proteomic complexity in PubMed DOI PMC

Kotsyfakis M., Schwarz A., Erhart J. & Ribeiro J. M. Tissue- and time-dependent transcription in PubMed DOI PMC

Xu X. L., Cheng T. Y., Yang H. & Liao Z. H. De novo assembly and analysis of midgut transcriptome of PubMed DOI

Richards S. A., Stutzer C., Bosman A. M. & Maritz-Olivier C. Transmembrane proteins–Mining the cattle tick transcriptome. Ticks and tick-borne diseases 6, 695–710, doi: 10.1016/j.ttbdis.2015.06.002 (2015). PubMed DOI

Krober T. & Guerin P. M. PubMed DOI

Perner J. et al. Acquisition of exogenous haem is essential for tick reproduction. Elife 5, doi: 10.7554/eLife.12318 (2016). PubMed DOI PMC

Lara F. A. et al. A new intracellular pathway of haem detoxification in the midgut of the cattle tick PubMed

Sojka D. et al. New insights into the machinery of blood digestion by ticks. Trends in parasitology 29, 276–285, doi: 10.1016/j.pt.2013.04.002 (2013). PubMed DOI

Lara F. A. Tracing heme in a living cell: hemoglobin degradation and heme traffic in digest cells of the cattle tick PubMed DOI

Schwarz A. et al. De novo PubMed DOI PMC

Perally S. r., LaCourse E. J., Campbell A. M. & Brophy P. M. Heme Transport and Detoxification in Nematodes: Subproteomics Evidence of Differential Role of Glutathione Transferases. Journal of Proteome Research 7, 4557–4565, doi: 10.1021/pr800395x (2008). PubMed DOI

Chisholm A. D. et al. Genome-Wide Analysis Reveals Novel Genes Essential for Heme Homeostasis in PubMed DOI PMC

Gulia-Nuss M. et al. Genomic insights into the PubMed DOI PMC

James M. O. & Ambadapadi S. Interactions of cytosolic sulfotransferases with xenobiotics. Drug metabolism reviews 45, 401–414, doi: 10.3109/03602532.2013.835613 (2013). PubMed DOI

Kim S.-J., Jung H.-J. & Lim C.-J. Reactive Oxygen Species-Dependent Down-Regulation of Tumor Suppressor Genes PTEN, USP28, DRAM, TIGAR, and CYLD Under Oxidative Stress. Biochemical Genetics 51, 901–915, doi: 10.1007/s10528-013-9616-7 (2013). PubMed DOI

Oliveira J. H. et al. Blood meal-derived heme decreases ROS levels in the midgut of PubMed DOI PMC

Bu C. et al. Novel and selective acetylcholinesterase inhibitors for PubMed DOI

Dermauw W. et al. A link between host plant adaptation and pesticide resistance in the polyphagous spider mite PubMed DOI PMC

Van Leeuwen T. & Dermauw W. The Molecular Evolution of Xenobiotic Metabolism and Resistance in Chelicerate Mites. Annu Rev Entomol 61, 475–498, doi: 10.1146/annurev-ento-010715-023907 (2016). PubMed DOI

Zhang Y., He Y., He L., Zong H.-Y. & Cai G.-B. Molecular cloning and characterization of a phospholipid hydroperoxide glutathione peroxidase gene from a blood fluke PubMed DOI

Claro da Silva T., Polli J. E. & Swaan P. W. The solute carrier family 10 (SLC10): beyond bile acid transport. Molecular aspects of medicine 34, 252–269, doi: 10.1016/j.mam.2012.07.004 (2013). PubMed DOI PMC

Maekawa S. et al. Analysis of RNA decay factor mediated RNA stability contributions on RNA abundance. BMC genomics 16, 154, doi: 10.1186/s12864-015-1358-y (2015). PubMed DOI PMC

Li J. J. & Biggin M. D. Gene expression. Statistics requantitates the central dogma. Science 347, 1066–1067, doi: 10.1126/science.aaa8332 (2015). PubMed DOI

Rokyta D. R., Margres M. J. & Calvin K. Post-transcriptional Mechanisms Contribute Little to Phenotypic Variation in Snake Venoms. G3 (Bethesda) 5, 2375–2382, doi: 10.1534/g3.115.020578 (2015). PubMed DOI PMC

Daran-Lapujade P. et al. The fluxes through glycolytic enzymes in PubMed DOI PMC

Kato N. et al. Regulatory mechanisms of chitin biosynthesis and roles of chitin in peritrophic matrix formation in the midgut of adult PubMed DOI

Narasimhan S. et al. Gut microbiota of the tick vector PubMed DOI PMC

Merzendorfer H. The cellular basis of chitin synthesis in fungi and insects: common principles and differences. Eur J Cell Biol 90, 759–769, doi: 10.1016/j.ejcb.2011.04.014 (2011). PubMed DOI

Lenardon M. D., Munro C. A. & Gow N. A. Chitin synthesis and fungal pathogenesis. Curr Opin Microbiol 13, 416–423, doi: 10.1016/j.mib.2010.05.002 (2010). PubMed DOI PMC

Quehenberger O. & Dennis E. A. The human plasma lipidome. N Engl J Med 365, 1812–1823, doi: 10.1056/NEJMra1104901 (2011). PubMed DOI PMC

Ding J. et al. Application of the accurate mass and time tag approach in studies of the human blood lipidome. J Chromatogr B Analyt Technol Biomed Life Sci 871, 243–252, doi: 10.1016/j.jchromb.2008.04.040 (2008). PubMed DOI PMC

Zhu K., Bowman A. S., Dillwith J. W. & Sauer J. R. Phospholipase A2 activity in salivary glands and saliva of the lone star tick (Acari: Ixodidae) during tick feeding. J Med Entomol 35, 500–504 (1998). PubMed

Zhu K., Dillwith J. W., Bowman A. S. & Sauer J. R. Identification of hemolytic activity in saliva of the lone star tick (Acari:Ixodidae). J Med Entomol 34, 160–166 (1997). PubMed

Kaufman W. R. Correlation between haemolymph ecdysteroid titre, salivary gland degeneration and ovarian development in the Ixodid tick, DOI

Friesen K. J. & Kaufman W. R. Quantification of vitellogenesis and its control by 20-hydroxyecdysone in the ixodid tick, Amblyomma hebraeum. Journal of Insect Physiology 48, 773–782, doi: 10.1016/s0022-1910(02)00107-5 (2002). PubMed DOI

Kaufman W. R. Gluttony and sex in female ixodid ticks: How do they compare to other blood-sucking arthropods? Journal of Insect Physiology 53, 264–273, doi: 10.1016/j.jinsphys.2006.10.004 (2007). PubMed DOI

Maya-Monteiro C. M. et al. HeLp, a Heme Lipoprotein from the Hemolymph of the Cattle Tick, PubMed DOI

Horn M. et al. Hemoglobin digestion in blood-feeding ticks: mapping a multipeptidase pathway by functional proteomics. Chemistry & biology 16, 1053–1063, doi: 10.1016/j.chembiol.2009.09.009 (2009). PubMed DOI PMC

Sojka D. et al. Profiling of proteolytic enzymes in the gut of the tick PubMed DOI PMC

Sojka D. et al. Multienzyme degradation of host serum albumin in ticks. Ticks and tick-borne diseases, doi: 10.1016/j.ttbdis.2015.12.014 (2015). PubMed DOI

Sojka D. et al. Characterization of gut-associated cathepsin D hemoglobinase from tick PubMed DOI PMC

Franta Z. et al. IrCL1-The haemoglobinolytic cathepsin L of the hard tick, PubMed

Tsuji N. et al. A cysteine protease is critical for PubMed DOI PMC

Motobu M. et al. Molecular characterization of a blood-induced serine carboxypeptidase from the ixodid tick PubMed DOI

Miyoshi T., Tsuji N., Islam M. K., Kamio T. & Fujisaki K. Cloning and molecular characterization of a cubilin-related serine proteinase from the hard tick PubMed DOI

Miyoshi T. et al. Molecular and reverse genetic characterization of serine proteinase-induced hemolysis in the midgut of the ixodid tick PubMed DOI

Pennington J. E., Goldstrohm D. A. & Wells M. A. The role of hemolymph proline as a nitrogen sink during blood meal digestion by the Mosquito PubMed DOI

Jenkinson C. P., Grody W. W. & Cederbaum S. D. Comparative properties of arginases. Comp Biochem Physiol B Biochem Mol Biol 114, 107–132 (1996). PubMed

Hamdy B. H. Biochemical and physiological studies of certain ticks (Ixodoidea). Excretion during ixodid feeding. J Med Entomol 14, 15–18 (1977). PubMed

Dusbabek F., Zahradnickova H. & Simek P. Chemical stability of assembly pheromone of argasid ticks (Ixodoidea: Argasidae). Folia Parasitologica 45, 62–66 (1998).

Grunclova L. et al. Two secreted cystatins of the soft tick PubMed DOI

Kobe B. et al. Isolation, Cloning and Structural Characterisation of Boophilin, a Multifunctional Kunitz-Type Proteinase Inhibitor from the Cattle Tick. PLoS ONE 3, e1624, doi: 10.1371/journal.pone.0001624 (2008). PubMed DOI PMC

Assumpcao T. C. et al. PubMed DOI PMC

Toledano M. B., Delaunay-Moisan A., Outten C. E. & Igbaria A. Functions and Cellular Compartmentation of the Thioredoxin and Glutathione Pathways in Yeast. Antioxidants & Redox Signaling 18, 1699–1711, doi: 10.1089/ars.2012.5033 (2013). PubMed DOI PMC

Prast-Nielsen S., Huang H.-H. & Williams D. L. Thioredoxin glutathione reductase: Its role in redox biology and potential as a target for drugs against neglected diseases. Biochimica et Biophysica Acta (BBA)-General Subjects 1810, 1262–1271, doi: 10.1016/j.bbagen.2011.06.024 (2011). PubMed DOI PMC

Cossío-Bayúgar R., Miranda E. & Holman P. J. Molecular cloning of a phospholipid-hydroperoxide glutathione peroxidase gene from the tick, PubMed DOI

Das S. et al. Salp25D, an PubMed DOI

Narasimhan S. et al. A Tick Antioxidant Facilitates the Lyme Disease Agent’s Successful Migration from the Mammalian Host to the Arthropod Vector. Cell Host Microbe 2, 7–18, doi: 10.1016/j.chom.2007.06.001 (2007). PubMed DOI PMC

McCord J. M. & Fridovich I. The utility of superoxide dismutase in studying free radical reactions. I. Radicals generated by the interaction of sulfite, dimethyl sulfoxide, and oxygen. J Biol Chem 244, 6056–6063 (1969). PubMed

Matoušková P., Vokřál I., Lamka J. & Skálová L. The Role of Xenobiotic-Metabolizing Enzymes in Anthelmintic Deactivation and Resistance in Helminths. Trends Parasitol, doi: 10.1016/j.pt.2016.02.004 (2016). PubMed DOI

Lambert L. A. Molecular evolution of the transferrin family and associated receptors. Biochim Biophys Acta 1820, 244–255, doi: 10.1016/j.bbagen.2011.06.002 ( 2012). PubMed DOI

Bettedi L., Aslam M. F., Szular J., Mandilaras K. & Missirlis F. Iron depletion in the intestines of Malvolio mutant flies does not occur in the absence of a multicopper oxidase. Journal of Experimental Biology 214, 971–978, doi: 10.1242/jeb.051664 (2011). PubMed DOI

Gunshin H. et al. Cloning and characterization of a mammalian proton-coupled metal-ion transporter. Nature 388, 482–488, doi: 10.1038/41343 (1997). PubMed DOI

Hajdusek O. et al. Tick iron and heme metabolism–New target for an anti-tick intervention. Ticks and Tick-borne Diseases, doi: 10.1016/j.ttbdis.2016.01.006 (2016). PubMed DOI

Hajdusek O. et al. Knockdown of proteins involved in iron metabolism limits tick reproduction and development. Proceedings of the National Academy of Sciences 106, 1033–1038, doi: 10.1073/pnas.0807961106 (2009). PubMed DOI PMC

Kopáček P. et al. Molecular cloning, expression and isolation of ferritins from two tick species— PubMed DOI

Tiklová K., Senti K.-A., Wang S., Gräslund A. & Samakovlis C. Epithelial septate junction assembly relies on melanotransferrin iron binding and endocytosis in PubMed DOI

Zhou G., Velasquez L. S., Geiser D. L., Mayo J. J. & Winzerling J. J. Differential regulation of transferrin 1 and 2 in PubMed DOI

Yuan X., Fleming M. D. & Hamza I. Heme transport and erythropoiesis. Current Opinion in Chemical Biology 17, 204–211, doi: 10.1016/j.cbpa.2013.01.010 (2013). PubMed DOI PMC

Korolnek T. & Hamza I. Like iron in the blood of the people: the requirement for heme trafficking in iron metabolism. Frontiers in Pharmacology 5, doi: 10.3389/fphar.2014.00126 (2014). PubMed DOI PMC

Shayeghi M. et al. Identification of an Intestinal Heme Transporter. Cell 122, 789–801, doi: 10.1016/j.cell.2005.06.025 (2005). PubMed DOI

Severance S. et al. Genome-wide analysis reveals novel genes essential for heme homeostasis in PubMed DOI PMC

Rajagopal A. et al. Haem homeostasis is regulated by the conserved and concerted functions of HRG-1 proteins. Nature 453, 1127–1131, doi: 10.1038/nature06934 (2008). PubMed DOI PMC

Quigley J. G. et al. Identification of a Human Heme Exporter that Is Essential for Erythropoiesis. Cell 118, 757–766, doi: 10.1016/j.cell.2004.08.014 (2004). PubMed DOI

Korolnek T., Zhang J., Beardsley S., Scheffer, George L. & Hamza I. Control of Metazoan Heme Homeostasis by a Conserved Multidrug Resistance Protein. Cell Metabolism 19, 1008–1019, doi: 10.1016/j.cmet.2014.03.030 (2014). PubMed DOI PMC

Michel K. et al. ATP Binding Cassette Transporter Mediates Both Heme and Pesticide Detoxification in Tick Midgut Cells. PLoS ONE 10, e0134779, doi: 10.1371/journal.pone.0134779 (2015). PubMed DOI PMC

Robinson M. D., McCarthy D. J. & Smyth G. K. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139–140, doi: 10.1093/bioinformatics/btp616 (2009). PubMed DOI PMC

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