Molecular genetic analysis of midgut serine proteases in Aedes aegypti mosquitoes

Isoe, Jun; Rascón, Alberto A.; Kunz, Susan; Miesfeld, Roger L.

doi:10.1016/j.ibmb.2009.10.008

Cited by 85 publications

(142 citation statements)

References 26 publications

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“…Mosquitoes had access to 3% sucrose-soaked cotton until the end of the experiments. Mortality and oviposition were monitored daily as previously described (22). The oviposition paper was removed 7 d after blood feeding, and the number of eggs laid by each female was counted.…”

Section: Egg Productionmentioning

confidence: 99%

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Effective disposal of nitrogen waste in blood‐fedAedes aegyptimosquitoes requires alanine aminotransferase

Mazzalupo¹,

Isoe²,

Belloni

et al. 2015

FASEB j.

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To better understand the mechanisms responsible for the success of female mosquitoes in their disposal of excess nitrogen, we investigated the role of alanine aminotransferase (ALAT) in blood-fed Aedes aegypti. Transcript and protein levels from the 2 ALAT genes were analyzed in sucrose-and blood-fed A. aegypti tissues. ALAT1 and ALAT2 exhibit distinct expression patterns in tissues during the first gonotrophic cycle. Injection of female mosquitoes with either double-stranded RNA (dsRNA)-ALAT1 or dsRNA ALAT2 significantly decreased mRNA and protein levels of ALAT1 or ALAT2 in fat body, thorax, and Malpighian tubules compared with dsRNA firefly luciferase-injected control mosquitoes. The silencing of either A. aegypti ALAT1 or ALAT2 caused unexpected phenotypes such as a delay in blood digestion, a massive accumulation of uric acid in the midgut posterior region, and a significant decrease of nitrogen waste excretion during the first 48 h after blood feeding. Concurrently, the expression of genes encoding xanthine dehydrogenase and ammonia transporter (Rhesus 50 glycoprotein) were significantly increased in tissues of both ALAT1-and ALAT2-deficient females. Moreover, perturbation of ALAT1 and ALAT2 in the female mosquitoes delayed oviposition and reduced egg production. These novel findings underscore the efficient mechanisms that blood-fed mosquitoes use to avoid ammonia toxicity and free radical damage.-Mazzalupo, S., Isoe, J., Belloni, V., Scaraffia, P. Y. Effective disposal of nitrogen waste in blood-fed Aedes aegypti mosquitoes requires alanine aminotransferase. FASEB J. 30, 111-120 (2016). www.fasebj.org

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Section: Egg Productionmentioning

confidence: 99%

“…To validate that digestion is disrupted by ALAT depletion through RNAi, we next evaluated the level of 3 major Ser proteases, 5G1, CxLT, and LT (22,23), in the midguts of dsRNA FL-and dsRNA ALAT1-injected females by Western blot analysis. As shown in Fig.…”

Section: Differential Alat1 and Alat2 Protein Expression In Mosquito mentioning

confidence: 99%

Effective disposal of nitrogen waste in blood‐fedAedes aegyptimosquitoes requires alanine aminotransferase

Mazzalupo¹,

Isoe²,

Belloni

et al. 2015

FASEB j.

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“…During blood feeding, ingested blood passes through the esophagus and the midgut anterior portion into the stomach, bypassing the crop (2). In the stomach, the blood forms a bolus surrounded by a chitin-protein envelope called the peritrophic matrix, within which the ingested blood undergoes digestion (33)(34)(35). Assayed 24 h PBM, the stomach of the wild-type mosquito female contained a compact dark brown bolus of digested blood, whereas its crop and the anterior midgut were void of any blood (Fig.…”

Section: Specific Antagomir Depletion Of Mir-275 Drastically Affects mentioning

confidence: 99%

microRNA miR-275 is indispensable for blood digestion and egg development in the mosquito Aedes aegypti

Bryant

Macdonald

Raikhel

2010

Proc. Natl. Acad. Sci. U.S.A.

161

164

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The mosquito Aedes aegypti is the major vector of arboviral diseases, particularly of Dengue fever, of which there are more than 100 million cases annually. Mosquitoes, such as A. aegypti , serve as vectors for disease pathogens because they require vertebrate blood for their egg production. Pathogen transmission is tightly linked to repeated cycles of obligatory blood feeding and egg maturation. Thus, the understanding of mechanisms governing egg production is necessary to develop approaches that limit the spread of mosquito-borne diseases. Previous studies have identified critical roles of hormonal- and nutrition-based target of rapamycin (TOR) pathways in controlling blood-meal–mediated egg maturation in mosquitoes. In this work, we uncovered another essential regulator of blood-meal–activated processes, the microRNA miR-275 . The depletion of this microRNA in A. aegypti females after injection of its specific antagomir resulted in severe defects in blood digestion, fluid excretion, and egg development, clearly demonstrating that miR-275 is indispensable for these physiological processes. miR-275 exhibits an expression profile that suggests its regulation by a steroid hormone, 20-hydroxyecdysone (20E). In vitro organ culture experiments demonstrated that miR-275 is induced by this hormone in the presence of amino acids, indicative of a dual regulation by 20E and TOR. This report has uncovered the critical importance of microRNAs in controlling blood-meal–activated physiological events required for completion of egg development in mosquito disease vectors.

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“…The two major classes of secreted proteases in the blood fed midgut are endoproteases, represented by trypsin-like and chymotrypsin-like serine proteases (SPs), and exopeptidases, that function as aminopeptidases and carboxypeptidases. 1 Blood digestion functions in a biphasic fashion, utilizing early and late phase digestive proteases that are specifically regulated at the transcriptional and post-transcriptional levels. In Aedes aegypti, the early trypsin (AaET) gene is expressed and its mRNA accumulated in midgut cells prior to blood feeding, while the synthesis of the AaET protein occurs soon after blood feeding.…”

Section: Introductionmentioning

confidence: 99%

“…In Aedes aegypti, the early trypsin (AaET) gene is expressed and its mRNA accumulated in midgut cells prior to blood feeding, while the synthesis of the AaET protein occurs soon after blood feeding. [1][2][3] Expression of the AaET gene is under the control of juvenile hormone (JH), 3 while the amino acid (AA)/target of rapamycin (TOR) pathway regulates AaET translation after blood feeding. 4 The gene encoding the chymotrypsin-like SP, JHA15, is also expressed in the Aedes midgut prior to blood feeding, and its expression is controlled by JH.…”

Section: Introductionmentioning

confidence: 99%

Mosquito-specific microRNA-1890 targets the juvenile hormone-regulated serine protease JHA15 in the female mosquito gut

et al. 2015

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S Raikhel (2015) Mosquito-specific microRNA-1890 targets the juvenile hormone-regulated serine protease JHA15 in the female mosquito gut, RNA Biology, 12:12, 1383-1390, DOI: 10.1080/15476286.2015 Females of the hematophagous mosquito species require a vertebrate blood meal to supply amino acids and other nutrients necessary for egg development, serving as the driving force for the spread of many vector-borne diseases in humans. Blood digestion utilizes both early and late phase serine proteases (SPs) that are differentially regulated at the transcriptional and post-transcriptional level. To uncover the regulatory complexity of SPs in the female mosquito midgut, we investigated involvement of miRNAs in regulating the juvenile hormone (JH)-controlled chymotrypsin-like SP, JHA15. We identified regulatory regions complementary to the mosquito-specific miRNA, miR-1890, within the 3 0 UTR of JHA15 mRNA. The level of the JHA15 transcript is highest post eclosion and drastically declines post blood meal (PBM), exhibiting an opposite trend to miR-1890 that peaks at 24 h PBM. Depletion of miR-1890 results in defects in blood digestion, ovary development and egg deposition. JHA15 mRNA and protein levels are elevated in female mosquitoes with miR-1890 inhibition. JHA15 RNA interference in the miR-1890 depletion background alleviates miR-1890 depletion phenotypes. The miR-1890 gene is activated by the 20-hydroxyecdysone pathway that involves the ecdysone receptor and the early genes, E74B and Broad Z2. Our study suggests that miR-1890 controls JHA15 mRNA stability in a stage-and tissue-specific manner.

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Molecular genetic analysis of midgut serine proteases in Aedes aegypti mosquitoes

Cited by 85 publications

References 26 publications

Effective disposal of nitrogen waste in blood‐fedAedes aegyptimosquitoes requires alanine aminotransferase

Effective disposal of nitrogen waste in blood‐fedAedes aegyptimosquitoes requires alanine aminotransferase

microRNA miR-275 is indispensable for blood digestion and egg development in the mosquito Aedes aegypti

Mosquito-specific microRNA-1890 targets the juvenile hormone-regulated serine protease JHA15 in the female mosquito gut

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