Food Ingestion and Digestive Enzymes in Larval Aedes aegypti and Ae. albopictus (Diptera: Culicidae)

Ho, Boon‐Chuan; Khoo, Hazel G.N.; Chew, Lita; Wong, Kim Ping; Ewert, A.

doi:10.1093/jmedent/29.6.960

Cited by 20 publications

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“…The band size range from ~25 to 250 kDa. The proteolytic activities of this mosquito have received little study, focusing mainly on larval activities and on comparisons with A. aegypti [38]. Comparisons between the previously reported peptidase profiles from the larval and pupal stages of A. albopictus [33] with the profile from the adult midgut obtained here reveals a similar banding pattern of enzymatic activity, suggesting that the peptidases expressed as part of the adult proteolytic machinery are already expressed in the pre-imaginal stages, where they mainly seem to play roles in digestion.…”

Section: Resultsmentioning

confidence: 99%

“…These activities were again strongly inhibited by PMSF and TLCK, but not by TPCK, (Figure 3B) corroborating the in-gel results. The occurrence of trypsin-like serine peptidases has been described in the larvae of A. aegypti and A. albopictus using both in-solution assays [38,39] and zymographic analysis [33,43]. The main peptidases detected in other Diptera species also belong to the serine peptidase class [42,44,45].…”

Section: Resultsmentioning

confidence: 99%

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The midgut of Aedes albopictus females expresses active trypsin-like serine peptidases

et al. 2014

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BackgroundAedes albopictus is widely distributed across tropical and sub-tropical regions and is associated with the transmission of several arboviruses. Although this species is increasingly relevant to public health due its ability to successfully colonize both urban and rural habitats, favoring the dispersion of viral infections, little is known about its biochemical traits, with all assumptions made based on studies of A. aegypti. In previous studies we characterized the peptidase profile of pre-imaginal stages of A. albopictus and we reported the first proteomic analysis of the midgut from sugar-fed females of this insect species.MethodsIn the present work, we further analyzed the peptidase expression in the midgut of sugar-fed females using 1DE-substrate gel zymography, two-dimensional electrophoresis (2DE), mass spectrometry (MS), and protein identification based on similarity.ResultsThe combination of zymography, in solution assays using fluorescent substrates and 2DE-MS/MS allowed us to identify the active serine peptidase “fingerprint” in the midgut of A. albopictus females. Zymographic analysis revealed a proteolytic profile composed of at least 13 bands ranging from ~25 to 250 kDa, which were identified as trypsin-like serine peptidases by using specific inhibitors of this class of enzymes. Concomitant use of the fluorogenic substrate Z-Phe-Arg-AMC and trypsin-like serine protease inhibitors corroborated the zymographic findings. Our proteomic approach allowed the identification of two different trypsin-like serine peptidases and one chymotrypsin in protein spots of the alkaline region in 2DE map of the A. albopictus female midgut. Identification of these protein coding genes was achieved by similarity to the A. aegypti genome sequences using Mascot and OMSSA search engines.ConclusionThese results allowed us to detect, identify and characterize the expression of active trypsin-like serine peptidases in the midgut of sugar-fed A. albopictus females. In addition, proteomic analysis allowed us to confidently assign the expression of two trypsin genes and one chymotrypsin gene to the midgut of this mosquito. These results contribute to the gene annotation in this species of unknown genome and represent a small but important step toward the protein-level functional and localization assignment of trypsin-like serine peptidase genes in the Aedes genus.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

The midgut of Aedes albopictus females expresses active trypsin-like serine peptidases

et al. 2014

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“…Earlier reports (Yang and Davies, 1971;Ho et. al., 1992; GenBank accession numbers AF487334, AF487426, and AY198134) have indicated that Ae aegypti larvae synthesize both trypsin and chymotrypsin-like enzymes.…”

Section: Synthesis Of Trypsin-and Chymotrypsin-like Enzymes In the MImentioning

confidence: 91%

“…However, its abundance in the adult CNS may indicate that the hormone has other functions. Yang and Davies (1971) and Ho et al (1992) reported that trypsin and chymotrypsin are the main digestive enzymes in Ae. aegypti.…”

Section: March 2004mentioning

confidence: 99%

“…aegypti when the hormone was fed with the blood meal. The potential of TMOF and its analogues as orally administered biorational insecticide "diet pills" to mosquito larvae that digest their food using trypsin-and chymotrypsin-like enzymes (Ho et al, 1992) prompted us to study the effect of orally ingested TMOF and its analogues on mosquito larval digestion, growth, and development. Furthermore, our earlier studies indicated that TMOF was synthesized and secreted in the adult ovaries following a blood meal (Borovsky et al, 1994b).…”

Section: Introductionmentioning

confidence: 99%

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Biochemical and cytoimmunological evidence for the control of Aedes aegypti larval trypsin with Aea‐TMOF

Borovsky

Meola

2004

Arch Insect Biochem Physiol

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Trypsin and chymotrypsin-like enzymes were detected in the gut of Aedes aegypti in the four larval instar and pupal developmental stages. Although overall the amount of trypsin synthesized in the larval gut was 2-fold higher than chymotrypsin, both enzymes are important in food digestion. Feeding Aea-Trypsin Modulating Oostatic Factor (TMOF) to Ae. aegypti and Culex quinquefasciatus larvae inhibited trypsin biosynthesis in the larval gut, stunted larval growth and development, and caused mortality. Aea-TMOF induced mortality in Ae. aegypti, Cx. quinquefasciatus, Culex nigripalpus, Anopheles quadrimaculatus, and Aedes taeniorhynchus larvae, indicating that many mosquito species have a TMOF-like hormone. The differences in potency of TMOF on different mosquito species suggest that analogues in other species are similar but may differ in amino acid sequence or are transported differently through the gut. Feeding of 29 different Aea-TMOF analogues to mosquito larvae indicated that full biological activity of the hormone is achieved with the tetrapeptide YDPA. Using cytoimmunochemical analysis, intrinsic TMOF was localized to ganglia of the central nervous system in larvae and male and female Ae. aegypti adults. The subesophageal, thoracic, and abdominal ganglia of both larval and adult mosquitoes contained immunoreactive cells. Immunoreactive cells were absent in the corpus cardiacum of newly molted 4th instar larvae but were found in late 4th instar larvae. In both males and females, the intrinsic neurosecretory cells of the corpus cardiacum were filled with densely stained immunoreactive material. These results indicate that TMOF-immunoreactive material is synthesized in sugar-fed male and female adults and larvae by the central nervous system cells.

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Expression of trypsin‐like serine peptidases in pre‐imaginal stages of Aedes aegypti (Diptera: Culicidae)

Mesquita-Rodrigues¹,

Sabóia-Vahia²,

Cuervo³

et al. 2011

Arch Insect Biochem Physiol

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This study reports the biochemical characterization and comparative analyses of highly active serine proteases in the larval and pupal developmental stages of Aedes aegypti (Linnaeus) using substrate‐SDS‐PAGE. Zymographic analysis of larval stadia detected proteolytic activity in 6–8 bands with apparent molecular masses ranging from 20 to 250 kDa, with activity observed from pH 5.5 to 10.0. The pupal stage showed a complex proteolytic activity in at least 11 bands with apparent Mr ranging from 25 to 250 kDa, and pH optimum at 10.0. The proteolytic activities of both larval and pupal stages were strongly inhibited by phenyl‐methyl sulfonyl‐fluoride and N‐α‐Tosyl‐L‐lysine chloromethyl ketone hydrochloride, indicating that the main proteases expressed by these developmental stages are trypsin‐like serine proteases. The enzymes were active at temperatures ranging from 4 to 85°C, with optimal activity between 37 and 60°C, and low activity at 85°C. Comparative analysis between the proteolytic enzymes expressed by larvae and pupae showed that substantial changes in the expression of active trypsin‐like serine proteases occur during the developmental cycle of A. aegypti. © 2011 Wiley Periodicals, Inc.

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Food Ingestion and Digestive Enzymes in Larval Aedes aegypti and Ae. albopictus (Diptera: Culicidae)

Cited by 20 publications

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The midgut of Aedes albopictus females expresses active trypsin-like serine peptidases

The midgut of Aedes albopictus females expresses active trypsin-like serine peptidases

Biochemical and cytoimmunological evidence for the control of Aedes aegypti larval trypsin with Aea‐TMOF

Expression of trypsin‐like serine peptidases in pre‐imaginal stages of Aedes aegypti (Diptera: Culicidae)

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