Synthesis, degradation and functions of insect peritrophic membrane

Author(s)

Renata Bolognesi

• Document type: Doctoral Thesis
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ)
• Defense date: 2005-04-05

Examining board members

Clélia Ferreira Terra; Pio Colepicolo Neto; Márcio de Castro Silva Filho; Carlos Eduardo Winter; Bianca Silvana Zingales

• Advisor: Clelia Ferreira Terra

Abstract

Most insects have a film-like anatomical structure (peritrophic membrane, PM) composed of chitin and proteins (peritrophins), which separates food from midgut tissue. It protects the epithelium against food abrasion and microrganisms and has other functions based on compartmentalization of enzymes, which include: increasing digestive efficiency by decreasing enzyme excretion and by other mechanisms that were tested in this thesis. The peritrophin synthesis is less known than PM chitin synthesis, which needs to be better understood. The characterization and expression of S. frugiperda genes encoding a peritrophin and enzymes responsible for the synthesis and degradation of chitin, chitin synthases 1(SfCHS1) and 2 (SfCHS2), and chitinase (SfCHI), respectively, were analysed. Sequences of corresponding cDNAs were determined by amplification of overlapping PCR fragments and the expression patterns of chitin metabolism genes were analyzed during insect development by RT-PCR. SfCHS2 is expressed in the midgut during the feeding stages, whereas SfCHI is expressed during the wandering and pupal stages. Both genes are predominantly expressed in the anterior portion of the midgut with a decreasing gradient of transcript levels in the medial and posterior portions. Chitin staining revealed that the polysaccharide is present in the PM only when SfCHS2 is expressed. There is little or no chitin in the midgut when SfCHI is expressed. These results support the hypothesis that SfCHS2 is responsible for PM chitin synthesis during the larval stage and SfCHI for PM chitin degradation during larval-pupal molting, suggesting inverse patterns of expression of these genes. The secretion site (anterior, middle or posterior midgut) of digestive enzymes can be predicted in Spodoptera frugiperda, Tenebrio molitor and Musca domestica based on enzyme activity distribution along the endoperitrophic space. We also have shown, using several experimental models, that the luminal compartment separation by PM: a) avoid the polimer hidrolases inhibition by removing oligomer from endoperitrophic space; b) decrease the oligomer hidrolases inhibition by restricting them to the ectoperitrophic space (by avoiding their contact with food); and c) block the inhibition of enzymes located at the cell surface involved in terminal digestion by avoiding their contact with food. (AU)