Abstract
The insect midgut epithelium is composed of columnar, goblet, and regenerative cells. Columnar epithelial cells are the most abundant and have membrane protrusions that form the brush border membrane (BBM) on their apical side. These increase surface area available for the transport of nutrients, but also provide opportunities for interaction with xenobiotics such as pathogens, toxins and host plant allelochemicals. Recent improvements in proteomic and bioinformatics tools provided an opportunity to determine the proteome of the T. ni BBM in unprecedented detail. This study reports the identification of proteins from BBM vesicles (BBMVs) using single dimension polyacrylamide gel electrophoresis coupled with multi‐dimensional protein identification technology. More than 3000 proteins were associated with the BBMV, of which 697 were predicted to possess either a signal peptide, at least one transmembrane domain or a GPI‐anchor signal. Of these, bioinformatics analysis and manual curation predicted that 185 may be associated with the BBMV or epithelial cell plasma membrane. These are discussed with respect to their predicted functions, namely digestion, nutrient uptake, cell signaling, development, cell–cell interactions, and other functions. We believe this to be the most detailed proteomic analysis of the lepidopteran midgut epithelium membrane to date, which will provide information to better understand the biochemical, physiological and pathological processes taking place in the larval midgut.
Highlights
IntroductionT. ni will feed on an exceptionally broad range of plants, including over 160 species in 36 families (Sutherland & Green, 1984)
BBM vesicles (BBMVs) were prepared from midgut tissue isolated from actively feeding 4th instar T. ni larvae
The T. ni BBMVs were observed by transmission electron microscopy after negative staining with phosphotungstic acid
Summary
T. ni will feed on an exceptionally broad range of plants, including over 160 species in 36 families (Sutherland & Green, 1984). Many of these plants are important crop species, including cruciferous vegetables. The gut is composed of three sections: the foregut, midgut and hindgut. The foregut is lined with a chitin-containing cuticle and facilitates physical processing of food. Posterior and anterior sphincters separate the midgut from the foregut and hindgut, respectively. The midgut is lined by the peritrophic matrix (PM), an acellular fibrous sheet composed mainly of chitin, mucopolysaccharides and proteins. The PM separates the food bolus from the midgut epithelial cell layer and prevents access by microorganisms and larger
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