title: Two essential peritrophic matrix proteins mediate matrix barrier functions in the insect midgut.
abstract: The peritrophic matrix (PM) in the midgut of insects consists primarily of chitin and proteins and is thought to support digestion and provide protection from abrasive food particles and enteric pathogens. We examined the physiological roles of 11 putative peritrophic matrix protein (PMP) genes of the red flour beetle, Tribolium castaneum (TcPMPs). TcPMP genes are differentially expressed along the length of the midgut epithelium of feeding larvae. RNAi of individual PMP genes revealed no abnormal developmental phenotypes for 9 of the 11 TcPMPs. However, RNAi for two PMP genes, TcPMP3 and TcPMP5-B, resulted in depletion of the fat body, growth arrest, molting defects and mortality. In situ permeability assays after oral administration of different-sized FITC-dextran beads demonstrated that the exclusion size of the larval peritrophic matrix (PM) decreases progressively from >2 MDa to <4 kDa from the anterior to the most posterior regions of the midgut. In the median midguts of control larvae, 2 MDa dextrans were completely retained within the PM lumen, whereas after RNAi for TcPMP3 and TcPMP5-B, these dextrans penetrated the epithelium of the median midgut, indicating loss of structural integrity and barrier function of the larval PM. In contrast, RNAi for TcPMP5-B, but not RNAi for TcPMP3, resulted in breakdown of impermeability to 4 and 40 kDa dextrans in the PM of the posterior midgut. These results suggest that specific PMPs are involved in the regulation of PM permeability, and that a gradient of barrier function is essential for survival and fat body maintenance.
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Kansas State University, 141 Chalmers Hall, Manhattan, KS 66506, USA.
authors.forename: Sinu
authors.initials: S
authors.lastname: Agrawal
authors.affiliation: Department of Biology, Chemistry, University of Osnabrück, Osnabrück 49069, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Kelkenberg
authors.affiliation: Department of Entomology, Kansas State University, 103 Waters Hall, Manhattan, KS 66506, USA.
authors.forename: Khurshida
authors.initials: K
authors.lastname: Begum
authors.affiliation: Department of Biology, Chemistry, University of Osnabrück, Osnabrück 49069, Germany.
authors.forename: Lea
authors.initials: L
authors.lastname: Steinfeld
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Kansas State University, 141 Chalmers Hall, Manhattan, KS 66506, USA.
authors.forename: Clay E
authors.initials: CE
authors.lastname: Williams
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Kansas State University, 141 Chalmers Hall, Manhattan, KS 66506, USA; Center for Grain and Animal Health Research, ARS-USDA, 1515 College Avenue, Manhattan, KS 66502, USA.
authors.forename: Karl J
authors.initials: KJ
authors.lastname: Kramer
authors.affiliation: Center for Grain and Animal Health Research, ARS-USDA, 1515 College Avenue, Manhattan, KS 66502, USA.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Beeman
authors.affiliation: Department of Entomology, Kansas State University, 103 Waters Hall, Manhattan, KS 66506, USA.
authors.forename: Yoonseong
authors.initials: Y
authors.lastname: Park
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Kansas State University, 141 Chalmers Hall, Manhattan, KS 66506, USA. Electronic address: smk@ksu.edu.
authors.forename: Subbaratnam
authors.initials: S
authors.lastname: Muthukrishnan
authors.affiliation: Department of Biology, Chemistry, University of Osnabrück, Osnabrück 49069, Germany. Electronic address: merzendorfer@biologie.uni-osnabrueck.de.
authors.forename: Hans
authors.initials: H
authors.lastname: Merzendorfer
mesh_terms: D000818:Animals; D004064:Digestive System / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D018978:Molting; D014231:Tribolium / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
chemical_list: D019476:Insect Proteins
keywords: Chitin; Dextrans; Fat body; Insect; Midgut; Peritrophic matrix proteins; Permeability; RNA interference; Red flour beetle; Tribolium castaneum