title: Effects of different dietary conditions on the expression of trypsin- and chymotrypsin-like protease genes in the digestive system of the migratory locust, Locusta migratoria.
abstract: While technological advancements have recently led to a steep increase in genomic and transcriptomic data, and large numbers of protease sequences are being discovered in diverse insect species, little information is available about the expression of digestive enzymes in Orthoptera. Here we describe the identification of Locusta migratoria serine protease transcripts (cDNAs) involved in digestion, which might serve as possible targets for pest control management. A total of 5 putative trypsin and 15 putative chymotrypsin gene sequences were characterized. Phylogenetic analysis revealed that these are distributed among 3 evolutionary conserved clusters. In addition, we have determined the relative gene expression levels of representative members in the gut under different feeding conditions. This study demonstrated that the transcript levels for all measured serine proteases were strongly reduced after starvation. On the other hand, larvae of L. migratoria displayed compensatory effects to the presence of Soybean Bowman Birk (SBBI) and Soybean Trypsin (SBTI) inhibitors in their diet by differential upregulation of multiple proteases. A rapid initial upregulation was observed for all tested serine protease transcripts, while only for members belonging to class I, the transcript levels remained elevated after prolonged exposure. In full agreement with these results, we also observed an increase in proteolytic activity in midgut secretions of locusts that were accustomed to the presence of protease inhibitors in their diet, while no change in sensitivity to these inhibitors was observed. Taken together, this paper is the first comprehensive study on dietary dependent transcript levels of proteolytic enzymes in Orthoptera. Our data suggest that compensatory response mechanisms to protease inhibitor ingestion may have appeared early in insect evolution.
authors.affiliation: Department of Animal Physiology and Neurobiology, Zoological Institute K.U.Leuven, Naamsestraat 59, B-3000 Leuven, Belgium. Electronic address: jorntspit@hotmail.com.
authors.forename: Jornt
authors.initials: J
authors.lastname: Spit
authors.affiliation: Department of Animal Physiology and Neurobiology, Zoological Institute K.U.Leuven, Naamsestraat 59, B-3000 Leuven, Belgium.
authors.forename: Sven
authors.initials: S
authors.lastname: Zels
authors.affiliation: Department of Animal Physiology and Neurobiology, Zoological Institute K.U.Leuven, Naamsestraat 59, B-3000 Leuven, Belgium.
authors.forename: Senne
authors.initials: S
authors.lastname: Dillen
authors.affiliation: Department of Animal Physiology and Neurobiology, Zoological Institute K.U.Leuven, Naamsestraat 59, B-3000 Leuven, Belgium.
authors.forename: Michiel
authors.initials: M
authors.lastname: Holtof
authors.affiliation: Department of Animal Physiology and Neurobiology, Zoological Institute K.U.Leuven, Naamsestraat 59, B-3000 Leuven, Belgium.
authors.forename: Niels
authors.initials: N
authors.lastname: Wynant
authors.affiliation: Department of Animal Physiology and Neurobiology, Zoological Institute K.U.Leuven, Naamsestraat 59, B-3000 Leuven, Belgium.
authors.forename: Jozef
authors.initials: J
authors.lastname: Vanden Broeck
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002918:Chymotrypsin / Q000378:metabolism; D004032:Diet; D041981:Gastrointestinal Tract; D015870:Gene Expression; D007814:Larva; D048188:Locusta migratoria / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D011480:Protease Inhibitors / Q000494:pharmacology; D057057:Serine Proteases / Q000235:genetics* / Q000378:metabolism; D013217:Starvation; D014357:Trypsin / Q000378:metabolism
chemical_list: D011480:Protease Inhibitors; D057057:Serine Proteases; D002918:Chymotrypsin; D014357:Trypsin
keywords: Diet; Digestion; Inhibitor; Insect; Protease; Proteolytic; Serine; Starvation