title: The peritrophic membrane of Spodoptera frugiperda: secretion of peritrophins and role in immobilization and recycling digestive enzymes.
abstract: A peritrophin from the Spodoptera frugiperda peritrophic membrane (PM) and microvillar proteins from S. frugiperda anterior midgut cells were isolated and used to raise antibodies in a rabbit. These antibodies, as well as a Tenebrio molitor amylase antibody that cross-reacts with S. frugiperda amylases, and wheat-germ aglutinin were used in immunolocalization experiments performed with the aid of confocal fluorescence and immunogold techniques. The results showed that the peritrophin was secreted by anterior midgut columnar cells in vesicles pinched-off the microvilli (microapocrine secretion). The resulting double membrane vesicles become single membrane vesicles by membrane fusion, releasing peritrophin and part of the amylase and trypsin. The remaining membranes still containing microvillar proteins and membrane-bound amylase and trypsin are incorporated into a jelly-like material associated with PM. Calcofluor-treated larvae lacking a PM were shown to lose the decreasing gradient of trypsin and chymotrypsin observed along the midgut of control larvae. This gradient is thought to be formed by a countercurrent flux of fluid (in the space between PM and midgut cells) that powers enzyme recycling.
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: R
authors.initials: R
authors.lastname: Bolognesi
authors.forename: A F
authors.initials: AF
authors.lastname: Ribeiro
authors.forename: W R
authors.initials: WR
authors.lastname: Terra
authors.forename: C
authors.initials: C
authors.lastname: Ferreira
mesh_terms: D000681:Amylases / Q000378:metabolism; D000818:Animals; D004064:Digestive System / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D005260:Female; D019476:Insect Proteins / Q000302:isolation & purification / Q000378:metabolism*; D007814:Larva; D008297:Male; D008562:Membrane Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D008871:Microvilli / Q000378:metabolism; D018411:Spodoptera / Q000378:metabolism*
chemical_list: D019476:Insect Proteins; D008562:Membrane Glycoproteins; D000681:Amylases; D010450:Endopeptidases