The midgut transcriptome of Lutzomyia longipalpis: comparative analysis of cDNA libraries from sugar-fed, blood-fed, post-digested and Leishmania infantum chagasi-infected sand flies.
The midgut transcriptome of Lutzomyia longipalpis: comparative analysis of cDNA libraries from sugar-fed, blood-fed, post-digested and Leishmania infantum chagasi-infected sand flies.
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DOI：
10.1186/1471-2164-9-15
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发表时间：
2008-01-14
期刊：
影响因子：
4.4
通讯作者：
Valenzuela JG
中科院分区：
文献类型：
--
作者：
Jochim RC;Teixeira CR;Laughinghouse A;Mu J;Oliveira F;Gomes RB;Elnaiem DE;Valenzuela JG
在沙蝇消化道内利什曼原虫的生活史中，寄生虫必须在血粉消化的恶劣环境中生存，逃离血液团注，附着在中肠上皮上，然后分化为感染性的半周期阶段。利什曼原虫与沙蝇肠道之间的分子相互作用知之甚少。在本工作中，我们对从沙蝇Lutzmyia long gipalpis的中肠组织中构建的五个cDNA文库进行了测序，并分析了在存在和不存在婴儿利什曼原虫的情况下，糖摄取、血液摄取以及血粉加工和排泄后的转录本。对糖喂养和血液喂养的cDNA文库的转录本进行比较分析，鉴定出在血液喂养过程中差异表达的转录本。这包括四个不同的微绒毛样蛋白(LuloMVP1、2、4和5)、两个周毛膜样蛋白、一个胰蛋白酶样蛋白(Lltryp1)、两个类糜蛋白酶蛋白(LuloChym1a和LuloChym2)和一个未知蛋白。通过血液喂养下调的转录本是微绒毛样蛋白(LuloMVP3)、胰酶样蛋白(Lltryp2)和ASTIN样金属蛋白酶(LuloAsta In)。此外，对血液喂养和利什曼原虫感染的中肠cdna文库进行了比较分析，发现了由于沙蝇肠道中存在利什曼原虫而存在差异表达的转录本。这包括下调的转录本，如四个微绒毛样蛋白(LuloMVP 1，2，4和5)，一个乳糜蛋白酶(LuloChym1A)和一个羧基肽酶(LuloCPepA1)等。在利什曼原虫存在的情况下，上调的中肠转录本是一种周福蛋白样蛋白(LuloPer1)，一种胰酶样蛋白(Lltryp2)和一种未知蛋白。这一转录组分析代表了从特定沙蝇组织中报告的最大一组序列数据，并提供了沙蝇Lutzmyia long gipalpis中存在的转录本的进一步信息。这一分析提供了这种沙蝇中肠中存在的分子的详细信息，以及可能受到血液摄食和利什曼原虫存在的影响的转录本。更重要的是，这一分析表明，婴儿利什曼原虫在血粉消化过程中改变了沙蝇中肠某些转录物的表达谱，这种调节可能与寄生虫在苍蝇肠道中的生存和建立有关。此外，这一分析表明，这些变化可能发生在消化血餐的过程中，而不是之后。
In the life cycle of Leishmania within the alimentary canal of sand flies the parasites have to survive the hostile environment of blood meal digestion, escape the blood bolus and attach to the midgut epithelium before differentiating into the infective metacyclic stages. The molecular interactions between the Leishmania parasites and the gut of the sand fly are poorly understood. In the present work we sequenced five cDNA libraries constructed from midgut tissue from the sand fly Lutzomyia longipalpis and analyzed the transcripts present following sugar feeding, blood feeding and after the blood meal has been processed and excreted, both in the presence and absence of Leishmania infantum chagasi. Comparative analysis of the transcripts from sugar-fed and blood-fed cDNA libraries resulted in the identification of transcripts differentially expressed during blood feeding. This included upregulated transcripts such as four distinct microvillar-like proteins (LuloMVP1, 2, 4 and 5), two peritrophin like proteins, a trypsin like protein (Lltryp1), two chymotrypsin like proteins (LuloChym1A and 2) and an unknown protein. Downregulated transcripts by blood feeding were a microvillar-like protein (LuloMVP3), a trypsin like protein (Lltryp2) and an astacin-like metalloprotease (LuloAstacin). Furthermore, a comparative analysis between blood-fed and Leishmania infected midgut cDNA libraries resulted in the identification of the transcripts that were differentially expressed due to the presence of Leishmania in the gut of the sand fly. This included down regulated transcripts such as four microvillar-like proteins (LuloMVP1,2, 4 and 5), a Chymotrypsin (LuloChym1A) and a carboxypeptidase (LuloCpepA1), among others. Upregulated midgut transcripts in the presence of Leishmania were a peritrophin like protein (LuloPer1), a trypsin-like protein (Lltryp2) and an unknown protein. This transcriptome analysis represents the largest set of sequence data reported from a specific sand fly tissue and provides further information of the transcripts present in the sand fly Lutzomyia longipalpis. This analysis provides the detailed information of molecules present in the midgut of this sand fly and the transcripts potentially modulated by blood feeding and by the presence of the Leishmania parasite. More importantly, this analysis suggests that Leishmania infantum chagasi alters the expression profile of certain midgut transcripts in the sand fly during blood meal digestion and that this modulation may be relevant for the survival and establishment of the parasite in the gut of the fly. Moreover, this analysis suggests that these changes may be occurring during the digestion of the blood meal and not afterwards.
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