Dataset Information

Triatoma sordida salivary glands LC-MSMS

ABSTRACT: Triatoma sordida salivary glands secrete a number of complex bioactive proteins during feeding that affect hemostatic and immunological systems from vertebrate hosts. In this study, salivary glands and saliva were collected from 5th instar nymphs and adult triatomines. Liquid chromatography tandem-mass spectrometry (LC-MS/MS) was used to identify T. sordida salivary proteins. A total of 133 proteins was identified. Lipocalins, Hemiptera specific families, CRiSP/Antigen-5 and Kazal-type protein inhibitors proteins match to 42.01%, 14.32%, 5.86% and 1.91% of the total spectrum count, respectively. These proteins are involved in several physiological roles, including anti-coagulant, immunity and anti-microorganism. Salivary proteins contribute to successful feeding.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Triatoma Sordida

TISSUE(S): Saliva

SUBMITTER: Carla Araújo

LAB HEAD: Sèbastien Olivier Charneau

PROVIDER: PXD028350 | Pride | 2022-02-15

REPOSITORIES: Pride

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Publications

An Integrative Sialomic Analysis Reveals Molecules From <i>Triatoma sordida</i> (Hemiptera: Reduviidae).

Praça Yanna Reis YR Santiago Paula Beatriz PB Charneau Sébastien S Mandacaru Samuel Coelho SC Bastos Izabela Marques Dourado IMD Bentes Kaio Luís da Silva KLDS Silva Sofia Marcelino Martins SMM da Silva Waldeyr Mendes Cordeiro WMC da Silva Ionizete Garcia IG de Sousa Marcelo Valle MV Soares Célia Maria de Almeida CMA Ribeiro José Marcos Chaves JMC Santana Jaime Martins JM de Araújo Carla Nunes CN

Frontiers in cellular and infection microbiology 20220103

Triatomines have evolved salivary glands that produce versatile molecules with various biological functions, including those leading their interactions with vertebrate hosts' hemostatic and immunological systems. Here, using high-throughput transcriptomics and proteomics, we report the first sialome study on the synanthropic triatomine <i>Triatoma sordida</i>. As a result, 57,645,372 reads were assembled into 26,670 coding sequences (CDS). From these, a total of 16,683 were successfully annotate ...[more]

PMID: 35047420

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Project description:In addressing R. microplus - A. marginale interactions, we propose and test three linked hypotheses. The first is that the tick gene response is organ specific: the midgut gene regulation is unique during feeding and during acquisition of A. marginale as compared to the salivary gland. This distinction is relevant as the two organs serve very different roles in the transmission biology of A. marginale with early survival and replication within the midgut epithelium, composed of highly phagocytic cells, required for initial colonization while a second round of replication in the salivary gland acini, composed of highly secretory cells, is required for transmission of an infectious dose in the saliva. Importantly, both the midgut epithelium and salivary glands have been identified as separate and distinct barriers for transmission of A. marginale and thus represent two potential sites where transmission could be blocked. The second hypothesis to be tested is that the salivary gland transcriptome is temporally dynamic. Initiation of tick attachment and feeding involves secretion of a virtual pharmacopeia including lytic enzymes, anticoagulants, and inhibitors of the mammalian innate immune and nocioceptive systems. Concomitantly, the acini provide an environment where A. marginale replicates >100 fold and are secreted into the saliva. Prior studies show that duration of feeding is a critical component of transmission efficiency, with increased efficiency positively correlated with time of tick feeding. The third hypothesis to be tested is that A. marginale colonization does not significantly modulate the tick midgut and salivary gland transcriptome. This hypothesis is based on observations by ourselves and others that tick infection does not impart a significant fitness cost on the vector. This is in contrast to other bacterial and protozoal pathogens that have dramatic effects on success of tick attachment, engorgement, and survival. A. marginale, similar to other tick-borne pathogens in the Family Anaplasmataceeae, is believed to have evolved from an arthropod-specific bacterium with relatively late adaptation to specific niches in mammalian hosts. Consequently, we predict that A. marginale is well adapted to its tick vector and utilizes the normal signaling pathways of the feeding tick with few, if any, effects on the midgut and salivary gland transcriptome. In this manuscript, we report the testing of these three hypotheses and present the results in context of the vector-pathogen-mammalian host interaction at the time of transmission. A Roche NimbleGen high-density gene expression microarray was custom designed based on the expressed sequence tag (EST) database, B. microplus Gene Index Version 2 (BmiGI V2) for R. microplus. The expression level of 14,447 R. microplus genes was analyzed from total RNA extracted from 10 different tick tissue samples; 30 arrays were included since triplicates of each different sample were analyzed as follow: unfed (midgut and salivary glands), blood feeding (2 days midgut and 2, 6 and 9 days salivary glands), A. marginale-infected blood feeding (2 days midgut and 2, 6 and 9 days salivary glands).

Project description:Ticks are obligate blood feeding ectoparasites that transmit a wide variety of pathogenic microorganisms to their vertebrate hosts. Amblyomma sculptum is vector of Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever, the most lethal rickettsiosis that affects humans. It is known that the transmission of pathogens by ticks is mainly associated with the physiology of the feeding process. Pathogens that are acquired with the blood meal must first colonize the tick gut and later the salivary glands in order to be transmitted during a subsequent blood feeding via saliva. Tick saliva contains a complex mixture of bioactive molecules with anticlotting, antiplatelet aggregation, vasodilatory, anti-inflammatory, and immunomodulatory properties to counteract both the hemostasis and defense mechanisms of the host. Besides facilitating tick feeding, the properties of saliva may also benefits survival and establishment of pathogens in the host. In the current study, we compared the sialotranscriptome of unfed A. sculptum ticks and those fed for 72 h on rabbits using next generation RNA sequencing (RNA-seq). The total of reads obtained were assembled in 9,560 coding sequences (CDSs) distributed in different functional classes. CDSs encoding secreted proteins, including lipocalins, mucins, protease inhibitors, glycine-rich proteins, metalloproteases, 8.9 kDa superfamily members, and immunity-related proteins were mostly upregulated by blood feeding. Selected CDSs were analyzed by real-time quantitative polymerase chain reaction preceded by reverse transcription (RT-qPCR), corroborating the transcriptional profile obtained by RNA-seq. Finally, high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis revealed 124 proteins in saliva of ticks fed for 96-120 h. The corresponding CDSs of 59 of these proteins were upregulated in salivary glands of fed ticks. To the best of our knowledge, this is the first report on the proteome of A. sculptum saliva. The functional characterization of the identified proteins might reveal potential targets to develop vaccines for tick control and/or blocking of R. rickettsii transmission as well as pharmacological bioproducts with antihemostatic, anti-inflammatory and antibacterial activities.

Dataset Information

Triatoma sordida salivary glands LC-MSMS

Publications

An Integrative Sialomic Analysis Reveals Molecules From <i>Triatoma sordida</i> (Hemiptera: Reduviidae).

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