Characterization of constitutive and putative differentially expressed mRNAs by means of expressed sequence tags, differential display reverse transcriptase-PCR and randomly amplified polymorphic DNA-PCR from the sand fly vector Lutzomyia longipalpis

Ramalho-Ortigão, JM; Temporal, P.; Oliveira, Smp de; Barbosa, A. C. O.; Vilela, ML; Rangel, Elizabeth Ferreira; Rp, Brazil; Traub-Cseko, Y. M.

doi:10.1590/s0074-02762001000100012

Cited by 28 publications

(26 citation statements)

References 37 publications

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“…Very little is known about the signaling cascades initiated after exposure to MAMPs in these sand flies, but the collection of putative immune genes described so far (26,(32)(33)(34) suggests that Lutzomyia is similar to Anopheles in the way it is regulating NF-B activation of signaling cascades. We identified one sequence in the L. longipalpis cDNA libraries (26) that was similar to the Drosophila Caspar gene sharing higher similarity to the A. gambiae Caspar sequence.…”

Section: Discussionmentioning

confidence: 99%

Caspar-like Gene Depletion Reduces Leishmania Infection in Sand Fly Host Lutzomyia longipalpis

Telleria

Sant’Anna

Ortigão-Farias

et al. 2012

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 99%

Caspar-like Gene Depletion Reduces Leishmania Infection in Sand Fly Host Lutzomyia longipalpis

Telleria

Sant’Anna

Ortigão-Farias

et al. 2012

Journal of Biological Chemistry

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“…Three expression libraries were constructed using cDNA made with RNAs purified from midguts dissected after 6 and 72 h post-blood meal, and 72 h after artificial infection with L. chagasi as described previously (Ramalho-Ortigao et al, 2001) using a Lambda Zap vector kit (Stratagene, La Jolla, CA). After excision, inserts were sequenced at the PDTIS/FIOCRUZ sequencing platform in an Applied Biosystems 3730 DNA Analyzer.…”

Section: Est Sequencingmentioning

confidence: 99%

Constitutive and blood meal‐induced trypsin genes in Lutzomyia longipalpis

Telleria

Pitaluga

Ortigão-Farias

et al. 2007

Arch Insect Biochem Physiol

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Trypsins constitute some of the most abundant midgut digestive proteases expressed by hematophagous insects upon blood feeding. In addition to their role in the digestion of the blood meal, these proteases also have been implicated in the ability of certain pathogens to infect their natural vector. In sand flies, digestive proteases including trypsins were associated with early killing of Leishmania and are believed to play a role in the species-specificity dictating sand fly vectorial capacity. Our group is involved in studies of midgut digestive proteases in the sand fly Lutzomyia longipalpis, the principal vector of visceral leishmaniasis in Brazil. Here we report on the identification of two cDNAs, Lltryp1 and Lltryp2, which code for putative midgut trypsins in L. longipalpis. Analyses of RNA abundance using semi-quantitative RT-PCR show a different pattern of expression between the two genes. Lltryp1 expression remains undetected until blood feeding and reaches a peak at 12 h post-blood meal (PBM), returning to pre-blood meal levels at 72 h PBM. Additionally, Lltryp1 expression is undetected during larval development. Lltryp2, on the other hand, is constitutively expressed as high levels in the non-blood fed female, but is reduced upon blood feeding. At the end of the digestive cycle, Lltryp2 regains its pre-blood meal levels. This cDNA also is present in all developmental stages and in adult males. This pattern of expression is reminiscent of what is seen in mosquitoes and Old World sand flies, but has characteristics that are unique to L. longipalpis.

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“…Previous studies from another sandXy, Phlebotomus papatasi, focused on the characterization of midgut molecules (Kamhawi et al 2004;Ramalho-Ortigão et al 2005) and the analysis of midgut ESTs (Ramalho-Ortigão et al 2007a). Regarding L. longipalpis ESTs, most of the available data are related to the whole insect (Ramalho-Ortigão et al 2001;Dillon et al 2006), salivary gland (Charlab et al 1999;Anderson et al 2006), and midgut (Jochim et al 2008). Our group investigated patterns of gene expression in L. longipalpis upon blood feeding and/or infection with L. i. chagasi, with special focus on the vector midgut, a very important tissue for insect physiology and vectorial competence.…”

Section: Introductionmentioning

confidence: 99%

EST sequencing of blood-fed and Leishmania-infected midgut of Lutzomyia longipalpis, the principal visceral leishmaniasis vector in the Americas

et al. 2009

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Leishmaniasis is an important worldwide public health problem. Visceral leishmaniasis caused by Leishmania infantum chagasi is mainly transmitted by Lutzomyia longipalpis in the Americas. Leishmania development within the sand fly vector is mostly restricted to the midgut. Thus, a comparative analysis of blood-fed versus infected midguts may provide an invaluable insight into various aspects of sand fly immunity, physiology of blood digestion, and, more importantly, of Leishmania development. To that end, we have engaged in a study to identify expressed sequenced tags (ESTs) from L. longipalpis cDNA libraries produced from midguts dissected at different times post blood meal and also after artificial infection with L. i. chagasi. A total of 2,520 ESTs were obtained and, according to the quality of the sequencing data obtained, assembled into 378 clusters and 1,526 individual sequences or singletons totalizing 1,904 sequences. Several sequences associated with defense, apoptosis, RNAi, and digestion processes were annotated. The data presented here increases current knowledge on the New World sand fly transcriptome, contributing to the understanding of various aspects of the molecular physiology of L. longipalpis, and mechanisms underlying the relationship of this sand fly species with L. i. chagasi.

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Characterization of constitutive and putative differentially expressed mRNAs by means of expressed sequence tags, differential display reverse transcriptase-PCR and randomly amplified polymorphic DNA-PCR from the sand fly vector Lutzomyia longipalpis

Cited by 28 publications

References 37 publications

Caspar-like Gene Depletion Reduces Leishmania Infection in Sand Fly Host Lutzomyia longipalpis

Caspar-like Gene Depletion Reduces Leishmania Infection in Sand Fly Host Lutzomyia longipalpis

Constitutive and blood meal‐induced trypsin genes in Lutzomyia longipalpis

EST sequencing of blood-fed and Leishmania-infected midgut of Lutzomyia longipalpis, the principal visceral leishmaniasis vector in the Americas

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