A Deep Insight into the Sialome of Male and Female Aedes aegypti Mosquitoes

Only adult female mosquitoes feed on blood, while both genders take sugar meals. Accordingly, several compounds associated with blood feeding (i.e. vasodilators, anti-clotting, anti-platelets) are found only in female glands, while enzymes associated with sugar feeding or antimicrobials (such as lysozyme) are found in the glands of both sexes. We performed de novo assembly of reads from adult Aedes aegypti female and male salivary gland libraries (285 and 90 million reads, respectively). By mapping back the reads to the assembled contigs, plus mapping the reads from a publicly available Ae. aegypti library from adult whole bodies, we identified 360 transcripts (including splice variants and alleles) overexpressed tenfold or more in the glands when compared to whole bodies. Moreover, among these, 207 were overexpressed fivefold or more in female vs. male salivary glands, 85 were near equally expressed and 68 were overexpressed in male glands. We call in particular the attention to C-type lectins, angiopoietins, female-specific Antigen 5, the 9.7 kDa, 12–14 kDa, 23.5 kDa, 62/34 kDa, 4.2 kDa, proline-rich peptide, SG8, 8.7 kDa family and SGS fragments: these polypeptides are all of unknown function, but due to their overexpression in female salivary glands and putative secretory nature they are expected to affect host physiology. We have also found many transposons (some of which novel) and several endogenous viral transcripts (probably acquired by horizontal transfer) which are overexpressed in the salivary glands and may play some role in tissue-specific gene regulation or represent a mechanism of virus interference. This work contributes to a near definitive catalog of male and female salivary gland transcripts from Ae. aegypti, which will help to direct further studies aiming at the functional characterization of the many transcripts with unknown function and the understanding of their role in vector-host interaction and pathogen transmission.

[1]  O. Marinotti,et al.  Apyrase and alpha-glucosidase in the salivary glands of Aedes albopictus. , 1996, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[2]  P. Nuttall,et al.  Inhibition of the antiviral action of interferon by tick salivary gland extract , 2000, Parasite immunology.

[3]  Pei-Yin Lim,et al.  Mosquito Saliva Causes Enhancement of West Nile Virus Infection in Mice , 2010, Journal of Virology.

[4]  Eleanor Howe,et al.  RNA-Seq analysis in MeV , 2011, Bioinform..

[5]  Xun Xu,et al.  SOAPdenovo-Trans: de novo transcriptome assembly with short RNA-Seq reads , 2013, Bioinform..

[6]  F. Estelle,et al.  Mosquito allergy and mosquito salivary allergens. , 2007, Protein and peptide letters.

[7]  A. James,et al.  The salivary glands of the vector mosquito, Aedes aegypti, express a novel member of the amylase gene family , 1993, Insect molecular biology.

[8]  G. Saccone,et al.  Genomic organization and splicing evolution of the doublesex gene, a Drosophila regulator of sexual differentiation, in the dengue and yellow fever mosquito Aedes aegypti , 2011, BMC Evolutionary Biology.

[9]  S. Higgs,et al.  Mosquito feeding-induced enhancement of Cache Valley Virus (Bunyaviridae) infection in mice. , 1998, Journal of medical entomology.

[10]  Günter P. Wagner,et al.  Measurement of mRNA abundance using RNA-seq data: RPKM measure is inconsistent among samples , 2012, Theory in Biosciences.

[11]  M. Lehane,et al.  The biology of blood-sucking in insects , 1991 .

[12]  Andrea Crisanti,et al.  A comprehensive gene expression atlas of sex- and tissue-specificity in the malaria vector, Anopheles gambiae , 2011, BMC Genomics.

[13]  Gautier Koscielny,et al.  VectorBase: improvements to a bioinformatics resource for invertebrate vector genomics , 2011, Nucleic Acids Res..

[14]  Jun Li,et al.  A mosquito‐specific protein family includes candidate receptors for malaria sporozoite invasion of salivary glands , 2006, Cellular microbiology.

[15]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[16]  H. Bellen,et al.  Discs Lost, a Novel Multi-PDZ Domain Protein, Establishes and Maintains Epithelial Polarity , 2003, Cell.

[17]  M. Crabtree,et al.  Genetic and phenotypic characterization of the newly described insect flavivirus, Kamiti River virus , 2003, Archives of Virology.

[18]  R. Dahl,et al.  Finding middle ground: environmental conflict resolution. , 2003, Environmental health perspectives.

[19]  J. Ribeiro,et al.  An updated catalogue of salivary gland transcripts in the adult female mosquito, Anopheles gambiae , 2005, Journal of Experimental Biology.

[20]  J. Ribeiro,et al.  Characterization of a vasodilator from the salivary glands of the yellow fever mosquito Aedes aegypti. , 1992, The Journal of experimental biology.

[21]  A. Spielman,et al.  Salivary apyrase of Aedes aegypti: characterization and secretory fate. , 1984, Comparative biochemistry and physiology. B, Comparative biochemistry.

[22]  J. Ribeiro,et al.  The salivary purine nucleosidase of the mosquito, Aedes aegypti. , 2003, Insect biochemistry and molecular biology.

[23]  M. O’Bryan,et al.  The CAP superfamily: cysteine-rich secretory proteins, antigen 5, and pathogenesis-related 1 proteins--roles in reproduction, cancer, and immune defense. , 2008, Endocrine reviews.

[24]  R. Tesh,et al.  Host Immune Response to Mosquito-Transmitted Chikungunya Virus Differs from That Elicited by Needle Inoculated Virus , 2010, PloS one.

[25]  J. Nagaraju,et al.  Doublesex: a conserved downstream gene controlled by diverse upstream regulators , 2010, Journal of Genetics.

[26]  J. Ribeiro,et al.  Toward a description of the sialome of the adult female mosquito Aedes aegypti. , 2002, Insect biochemistry and molecular biology.

[27]  O. Marinotti,et al.  Salivary gland proteins of the mosquito Culex quinquefasciatus. , 2000, Archives of insect biochemistry and physiology.

[28]  S. Yoshida,et al.  Characterization and identification of exflagellation-inducing factor in the salivary gland of Anopheles stephensi (Diptera: Culicidae). , 2001, Biochemical and biophysical research communications.

[29]  A. James,et al.  Diet and salivation in female Aedes aegypti mosquitoes , 1990 .

[30]  Waseem Akram,et al.  Spatial mapping of gene expression in the salivary glands of the dengue vector mosquito, aedes aegypti , 2011, Parasites & Vectors.

[31]  S. Higgs,et al.  Potentiation of West Nile encephalitis by mosquito feeding. , 2006, Viral immunology.

[32]  A. James,et al.  Characterization of the Sialokinin I gene encoding the salivary vasodilator of the yellow fever mosquito, Aedes aegypti , 1999, Insect molecular biology.

[33]  Xiaofang Jiang,et al.  Complete Dosage Compensation in Anopheles stephensi and the Evolution of Sex-Biased Genes in Mosquitoes , 2015, Genome biology and evolution.

[34]  R. Titus,et al.  The immunomodulatory factors of arthropod saliva and the potential for these factors to serve as vaccine targets to prevent pathogen transmission , 2006, Parasite immunology.

[35]  R. Martienssen,et al.  Transposable elements and the epigenetic regulation of the genome , 2007, Nature Reviews Genetics.

[36]  Inanç Birol,et al.  De novo transcriptome assembly with ABySS , 2009, Bioinform..

[37]  L. P. Lounibos,et al.  An insight into the sialome of the frog biting fly, Corethrella appendiculata. , 2014, Insect biochemistry and molecular biology.

[38]  A. James,et al.  The salivary gland-specific apyrase of the mosquito Aedes aegypti is a member of the 5'-nucleotidase family. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[39]  R. Sinden,et al.  Identification of xanthurenic acid as the putative inducer of malaria development in the mosquito , 1998, Nature.

[40]  J. Ribeiro,et al.  An insight into the sialotranscriptome of the non-blood feeding Toxorhynchites amboinensis mosquito. , 2008, Insect biochemistry and molecular biology.

[41]  Bacteriolytic factor in the salivary glands of Aedes aegypti. , 1986, Comparative biochemistry and physiology. B, Comparative biochemistry.

[42]  Roland Contreras,et al.  Human Antimicrobial Peptides: Defensins, Cathelicidins and Histatins , 2005, Biotechnology Letters.

[43]  J. Ribeiro,et al.  Sialokinin I and II: vasodilatory tachykinins from the yellow fever mosquito Aedes aegypti. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[44]  S. Pfaff,et al.  Transposable elements as genetic regulatory substrates in early development. , 2013, Trends in cell biology.

[45]  A. James,et al.  A factor Xa-directed anticoagulant from the salivary glands of the yellow fever mosquito Aedes aegypti. , 1995, Experimental parasitology.

[46]  P. Nuttall Molecular characterization of tick-virus interactions. , 2009, Frontiers in bioscience.

[47]  R. Charrel,et al.  Sequences of flavivirus-related RNA viruses persist in DNA form integrated in the genome of Aedes spp. mosquitoes. , 2004, The Journal of general virology.

[48]  R. Charlab,et al.  The salivary adenosine deaminase activity of the mosquitoes Culex quinquefasciatus and Aedes aegypti. , 2001, The Journal of experimental biology.

[49]  P. Sohnle,et al.  Histidine-based zinc-binding sequences and the antimicrobial activity of calprotectin. , 1998, The Journal of infectious diseases.

[50]  J. Ribeiro,et al.  A Deep Insight into the Sialotranscriptome of the Gulf Coast Tick, Amblyomma maculatum , 2011, PloS one.

[51]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[52]  P. Nuttall,et al.  Non-viraemic transmission of tick-borne encephalitis virus: a mechanism for arbovirus survival in nature , 1993, Experientia.

[53]  T. Lumley,et al.  gplots: Various R Programming Tools for Plotting Data , 2015 .

[54]  R. Rosenberg,et al.  Xanthurenic Acid Induces Gametogenesis in Plasmodium, the Malaria Parasite* , 1998, The Journal of Biological Chemistry.

[55]  K. Vernick,et al.  Members of the Salivary Gland Surface Protein (SGS) Family Are Major Immunogenic Components of Mosquito Saliva* , 2011, The Journal of Biological Chemistry.

[56]  J. Ribeiro,et al.  The sialotranscriptome of adult male Anopheles gambiae mosquitoes. , 2006, Insect biochemistry and molecular biology.

[57]  A. Spielman,et al.  Fluid transport across the ducts of the salivary glands of a mosquito , 1982 .

[58]  A deep insight into the sialotranscriptome of the mosquito, Psorophora albipes , 2013, BMC Genomics.

[59]  P. Rossignol,et al.  Age dependence of salivary bacteriolytic activity in adult mosquitoes. , 1990, Comparative biochemistry and physiology. B, Comparative biochemistry.

[60]  S. Higgs,et al.  The enhancement of arbovirus transmission and disease by mosquito saliva is associated with modulation of the host immune response. , 2008, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[61]  J. Ribeiro,et al.  An insight into the sialome of blood-feeding Nematocera. , 2010, Insect biochemistry and molecular biology.

[62]  S. Higgs,et al.  Aedes aegypti Saliva Alters Leukocyte Recruitment and Cytokine Signaling by Antigen-Presenting Cells during West Nile Virus Infection , 2010, PloS one.

[63]  J. Cory,et al.  Non-viraemic transmission of Thogoto virus: vector efficiency of Rhipicephalus appendiculatus and Amblyomma variegatum. , 1990, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[64]  A. James,et al.  Evidence for two distinct members of the amylase gene family in the yellow fever mosquito, Aedes aegypti. , 1997, Insect biochemistry and molecular biology.

[65]  Rebecca J. Oakey,et al.  Transposable Elements Re-Wire and Fine-Tune the Transcriptome , 2013, PLoS genetics.

[66]  J. Ribeiro,et al.  Toward a catalog for the transcripts and proteins (sialome) from the salivary gland of the malaria vector Anopheles gambiae. , 2002, The Journal of experimental biology.