Ixodes ricinus is a tick that transmits the pathogens of Lyme and several arboviral diseases. Pathogens invade the tick midgut, disseminate through the hemolymph, and are transmitted to the vertebrate host via the salivary glands; subverting these processes could be used to interrupt pathogen transfer. Here, we use massive de novo sequencing to characterize the transcriptional dynamics of the salivary and midgut tissues of nymphal and adult I. ricinus at various time points after attachment on the vertebrate host. Members of a number of gene families show stage- and time-specific expression. We hypothesize that gene expression switching may be under epigenetic control and, in support of this, identify 34 candidate proteins that modify histones. I. ricinus-secreted proteins are encoded by genes that have a non-synonymous to synonymous mutation rate even greater than immune-related genes. Midgut transcriptome (mialome) analysis reveals several enzymes associated with protein, carbohydrate, and lipid digestion, transporters and channels that might be associated with nutrient uptake, and immune-related transcripts including antimicrobial peptides. This publicly available dataset supports the identification of protein and gene targets for biochemical and physiological studies that exploit the transmission lifecycle of this disease vector for preventative and therapeutic purposes.

Institute of Parasitology Biology Center of the Czech Academy of Sciences Branisovska 31 37005 Budweis Czech Republic

Section of Vector Biology Laboratory of Malaria and Vector Research National Institute of Allergy and Infectious Diseases 12735 Twinbrook Parkway room 2E32D Rockville MD 20852 USA

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Mannelli A., Bertolotti L., Gern L. & Gray J. Ecology of Borrelia burgdorferi sensu lato in Europe: transmission dynamics in multi-host systems, influence of molecular processes and effects of climate change. FEMS microbiology reviews 36, 837–861, 10.1111/j.1574-6976.2011.00312.x (2012). PubMed DOI

Pettersson J. H., Golovljova I., Vene S. & Jaenson T. G. Prevalence of tick-borne encephalitis virus in Ixodes ricinus ticks in northern Europe with particular reference to Southern Sweden. Parasit Vectors 7, 102, 10.1186/1756-3305-7-102 (2014). PubMed DOI PMC

Francischetti I. M. B., Sá-Nunes A., Mans B. J., Santos I. M. & Ribeiro J. M. C. The role of saliva in tick feeding. Frontiers in Biosciences 14, 2051–2088 (2009). PubMed PMC

Schwarz A. et al. De novo Ixodes ricinus salivary gland transcriptome analysis using two next-generation sequencing methodologies. Faseb J 27, 4745–4756, 10.1096/fj.13-232140 (2013). PubMed DOI PMC

Chmelar J. et al. Insight into the sialome of the castor bean tick, Ixodes ricinus. BMC Genomics 9, 233 (2008). PubMed PMC

Schwarz A. et al. A systems level analysis reveals transcriptomic and proteomic complexity in Ixodes ricinus midgut and salivary glands during early attachment and feeding. Mol. Cel. Proteomics In Press (2014). PubMed PMC

Liu X. Y. et al. IrSPI, a Tick Serine Protease Inhibitor Involved in Tick Feeding and Bartonella henselae Infection. PLoS neglected tropical diseases 8, e2993, 10.1371/journal.pntd.0002993 (2014). PubMed DOI PMC

Rachinsky A., Guerrero F. D. & Scoles G. A. Proteomic profiling of Rhipicephalus (Boophilus) microplus midgut responses to infection with Babesia bovis. Veterinary parasitology 152, 294–313, 10.1016/j.vetpar.2007.12.027 (2008). PubMed DOI

Heekin A. M. et al. Gut transcriptome of replete adult female cattle ticks, Rhipicephalus (Boophilus) microplus, feeding upon a Babesia bovis-infected bovine host. Parasitology research 112, 3075–3090, 10.1007/s00436-013-3482-4 (2013). PubMed DOI

Francischetti I. M., Sa-Nunes A., Mans B. J., Santos I. M. & Ribeiro J. M. The role of saliva in tick feeding. Front Biosci 14, 2051–2088 (2009). PubMed PMC

Ribeiro J. M. et al. An annotated catalog of salivary gland transcripts from Ixodes scapularis ticks. Insect Biochem. Mol. Biol. 36, 111–129 (2006). PubMed

Ceraul S. M. et al. A Kunitz protease inhibitor from Dermacentor variabilis, a vector for spotted fever group rickettsiae, limits Rickettsia montanensis invasion. Infection and immunity 79, 321–329, 10.1128/IAI.00362-10 (2011). PubMed DOI PMC

Ceraul S. M., Dreher-Lesnick S. M., Mulenga A., Rahman M. S. & Azad A. F. Functional characterization and novel rickettsiostatic effects of a Kunitz-type serine protease inhibitor from the tick Dermacentor variabilis. Infection and immunity 76, 5429–5435, 10.1128/IAI.00866-08 (2008). PubMed DOI PMC

Anderson J. M., Sonenshine D. E. & Valenzuela J. G. Exploring the mialome of ticks: an annotated catalogue of midgut transcripts from the hard tick, Dermacentor variabilis (Acari: Ixodidae). BMC genomics 9, 552, 10.1186/1471-2164-9-552 (2008). PubMed DOI PMC

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

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

Mulenga A., Misao O. & Sugimoto C. Three serine proteinases from midguts of the hard tick Rhipicephalus appendiculatus; cDNA cloning and preliminary characterization. Experimental & applied acarology 29, 151–164 (2003). PubMed

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 Haemaphysalis longicornis. Insect biochemistry and molecular biology 34, 799–808, 10.1016/j.ibmb.2004.04.004 (2004). PubMed DOI

Matsuo T. et al. Morphological studies on the extracellular structure of the midgut of a tick, Haemaphysalis longicornis (Acari: Ixodidae). Parasitology research 90, 243–248, 10.1007/s00436-003-0833-6 (2003). PubMed DOI

Caperucci D., Bechara G. H. & Camargo Mathias M. I. Ultrastructure features of the midgut of the female adult Amblyomma cajennense ticks Fabricius, 1787 (Acari: Ixodidae) in several feeding stages and subjected to three infestations. Micron 41, 710–721, 10.1016/j.micron.2010.05.015 (2010). PubMed DOI

Ribeiro J. M. The midgut hemolysin of Ixodes dammini (Acari: Ixodidae). J. Parasitol. 74, 532–537 (1988). PubMed

de Parseval N. & Heidmann T. Human endogenous retroviruses: from infectious elements to human genes. Cytogenetic and genome research 110, 318–332, 10.1159/000084964 (2005). PubMed DOI

Inbar-Feigenberg M., Choufani S., Butcher D. T., Roifman M. & Weksberg R. Basic concepts of epigenetics. Fertility and sterility 99, 607–615, 10.1016/j.fertnstert.2013.01.117 (2013). PubMed DOI

Iovino N. Drosophila epigenome reorganization during oocyte differentiation and early embryogenesis. Briefings in functional genomics 13, 246–253, 10.1093/bfgp/elu007 (2014). PubMed DOI

Jiang L. et al. PfSETvs methylation of histone H3K36 represses virulence genes in Plasmodium falciparum. Nature 499, 223–227, 10.1038/nature12361 (2013). PubMed DOI PMC

Geraci N. S., Spencer Johnston J., Paul Robinson J., Wikel S. K. & Hill C. A. Variation in genome size of argasid and ixodid ticks. Insect biochemistry and molecular biology 37, 399–408 (2007). PubMed

Ullmann A. J., Lima C. M., Guerrero F. D., Piesman J. & Black W. C. T. Genome size and organization in the blacklegged tick, Ixodes scapularis and the Southern cattle tick, Boophilus microplus. Insect molecular biology 14, 217–222 (2005). PubMed

Chagas A. C. et al. A deep insight into the sialotranscriptome of the mosquito, Psorophora albipes. BMC genomics 14, 875, 10.1186/1471-2164-14-875 (2013). PubMed DOI PMC

Arca B. et al. Positive selection drives accelerated evolution of mosquito salivary genes associated with blood-feeding. Insect molecular biology 23, 122–131, 10.1111/imb.12068 (2014). PubMed DOI PMC

Kotsyfakis M., Karim S., Andersen J. F., Mather T. N. & Ribeiro J. M. Selective cysteine protease inhibition contributes to blood-feeding success of the tick Ixodes scapularis. The Journal of biological chemistry 282, 29256–29263 (2007). PubMed

Birol I. et al. De novo transcriptome assembly with ABySS. Bioinformatics (Oxford, England) 25, 2872–2877, 10.1093/bioinformatics/btp367 (2009). PubMed DOI

Simpson J. T. et al. ABySS: a parallel assembler for short read sequence data. Genome research 19, 1117–1123, 10.1101/gr.089532.108 (2009). PubMed DOI PMC

Yang Y. & Smith S. A. Optimizing de novo assembly of short-read RNA-seq data for phylogenomics. BMC genomics 14, 328, 10.1186/1471-2164-14-328 (2013). PubMed DOI PMC

Karim S., Singh P. & Ribeiro J. M. A deep insight into the sialotranscriptome of the Gulf Coast tick, Amblyomma maculatum. PLoS ONE 6, e28525, 10.1371/journal.pone.0028525 (2011). PubMed DOI PMC

Trapnell C. et al. Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nature protocols 7, 562–578, 10.1038/nprot.2012.016 (2012). PubMed DOI PMC

Li H. & Durbin R. Fast and accurate long-read alignment with Burrows-Wheeler transform. Bioinformatics (Oxford, England) 26, 589–595, 10.1093/bioinformatics/btp698 (2010). PubMed DOI PMC

Li H. et al. The Sequence Alignment/Map format and SAMtools. Bioinformatics (Oxford, England) 25, 2078–2079, 10.1093/bioinformatics/btp352 (2009). PubMed DOI PMC

Insight Into the Dynamics of the Ixodes ricinus Nymphal Midgut Proteome

Transcriptomic analysis of the tick midgut and salivary gland responses upon repeated blood-feeding on a vertebrate host

Haem-responsive gene transporter enables mobilization of host haem in ticks

Mialostatin, a Novel Midgut Cystatin from Ixodes ricinus Ticks: Crystal Structure and Regulation of Host Blood Digestion

Iripin-3, a New Salivary Protein Isolated From Ixodes ricinus Ticks, Displays Immunomodulatory and Anti-Hemostatic Properties In Vitro

The Central Role of Salivary Metalloproteases in Host Acquired Resistance to Tick Feeding

A combined transcriptomic approach to identify candidates for an anti-tick vaccine blocking B. afzelii transmission

Catalogue of stage-specific transcripts in Ixodes ricinus and their potential functions during the tick life-cycle

Sialome diversity of ticks revealed by RNAseq of single tick salivary glands

In silico target network analysis of de novo-discovered, tick saliva-specific microRNAs reveals important combinatorial effects in their interference with vertebrate host physiology

Protease Inhibitors in Tick Saliva: The Role of Serpins and Cystatins in Tick-host-Pathogen Interaction

Tick-Pathogen Interactions and Vector Competence: Identification of Molecular Drivers for Tick-Borne Diseases

RNA-seq analyses of the midgut from blood- and serum-fed Ixodes ricinus ticks

All For One and One For All on the Tick-Host Battlefield

Anaplasma phagocytophilum increases the levels of histone modifying enzymes to inhibit cell apoptosis and facilitate pathogen infection in the tick vector Ixodes scapularis

Acquisition of exogenous haem is essential for tick reproduction

Sialomes and Mialomes: A Systems-Biology View of Tick Tissues and Tick-Host Interactions

Modulation of host immunity by tick saliva

Deep Sequencing Analysis of the Ixodes ricinus Haemocytome