In order to understand the regulation of trypsin genes by the blood meal, we constructed a cDNA library from mRNA isolated from midguts of blood-fed female Aedes aegypti. The library was screened with a Drosophila melanogaster trypsin-like gene; twelve cDNAs were isolated and sequenced. Two clones were 846 bp and 788 bp long with 762 bp and 716 bp open reading frames, respectively. The cDNAs were identified as coding for serine proteases by the presence of conserved serine, histidine and aspartic acid residues; the presence of an aspartate residue at position 176 suggests that the clones were derived from trypsin-like gene transcripts rather than chymotrypsin or other serine proteases. One of the clones contained a 5' untranslated region and coding regions for putative signal and activation peptides, suggesting that the product is secreted as an inactive zymogen and processed by autoactivation. Southern analysis of genomic DNA suggests that trypsin is encoded by a multigene family in A. aegypti.

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Modifications of posterior midgut cells of Rhodnius prolixus following a meal of rabbit blood are described. Prominent stacks and whorls of rough endoplasmic reticulum become redistributed following a blood meal but later reform during the postfeeding period. Lysosomes undergo internal structural changes and apparent fluctuations in their number per cell as a result of blood meal ingestion. Before blood feeding, the apical surface of the midgut cells show a typical arrangement of a plasma membrane covered on the lumenal surface by a glycocalyx. After a blood meal, a more complex organisation appears, consisting of two plasma membranes separated by an electron-dense matrix. The lumenal apical membrane proliferates during the digestion period to form loosely organised extracellular membrane layers which may function as a peritrophic membrane. Changes in the rough endoplasmic reticulum and lysosomes and modifications to the apical cell surface appear to coincide with previously described proteinase activity cycles in the posterior midgut of R. prolixus. The implications of these results are discussed and are compared with similar ultrastructural events from haematophagous Diptera.

Protease activity in the alimentary tract of the European corn borer, Ostrinia nubilalis, can be attributed to at least three endoproteinases. A high alkaline trypsin with maximal hydrolysis of benzoyl arginine p-nitroanilide at pH values higher than 10.0, a low alkaline trypsin with maximal activity against benzoyl arginine ethyl ester at pH 9.0, and chymotrypsin, which hydrolyzed benzoyl tryosine ethyl ester at pH 7.5–8.0, were detected in gut homogenates. Total proteolysis, measured using azocasein, had maximal activity at pH 10.0 or higher. Corn borer chymotryptic activity had characteristics similar to the vertebrate enzyme. Both tryptic activities differed from vertebrate trypsin by being insensitive to ovomucoid trypsin inhibitor. High and low alkaline trypsin differed from each other by their pH optima. High alkaline trypsin was activated by magnesium and calcium, and low alkaline trypsin was not affected by inclusion of either chemical in the assay mixture.

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Continuous colonies of Chironomus riparius Meigen can be conveniently maintained in the laboratory and the swarming and mating behaviour is readily observable. Swarming is eocrepuscular and is mediated, at least in part, by light intensity and the marker stimulus. The effects of sound and volatile chemicals on the initiation and maintenance of swarming are not clearly apparent, but there is some evidence to suggest that pheromones may play a role in swarm cohesion.

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