Binding of the insecticidal lectin Concanavalin A in pea aphid, Acyrthosiphon pisum (Harris) and induced effects on the structure of midgut epithelial cells.
Concanavalin A (lectin from Canavalia ensiformis L., ConA) has previously been shown to act as a feeding inhibitor for Acyrthosiphon pisum, the pea aphid. In the present study a range of histochemical and biochemical techniques were used to elucidate the target tissues and binding sites of the lectin in the aphid. Diet uptake was evaluated using a radioactive tracer (14C-methylated inulin) and demonstrated that adults were capable of ingesting high quantities of the toxin (approx. 1 microg over a 48 h period). Electophoretic analysis and enzyme-linked immuno-sorbent assay of honeydew samples confirmed these results and further demonstrated that only small levels of ConA were excreted. Histofluorescence and immunolocalisation studies on nymphs revealed that the stomach was the primary target for ConA. At concentrations up to 400 microg ml(-1), lectin binding only occurred in the stomach region, however, at high concentrations (800 microg ml(-1)) the whole digestive tract was stained, although there was no evidence of binding in either the oesophagus or rectum. In addition to binding, there was evidence to suggest that ConA was also causing systemic effects in that the lectin appeared to cross the intestinal epithelial barrier. Immunohistochemical and electron microscopy studies revealed that ConA induced severe cellular swelling of the epithelial cells, accompanied by hypersecretion and a progressive detachment of the apical membrane; however, the striated border itself did not appear to be directly affected. Furthermore, there was no lysis of the epithelium, nor loss of integrity of the epithelial cells themselves. Our results suggest that ConA interacts with glycosylated receptors at the surface of the stomach epithelial cells, interfering with normal metabolism and cell function, resulting in a rapid feedback response on feeding behaviour. Whilst our results provide a much greater understanding regarding the modes of action of ConA in insects, they suggest that different lectins, including other mannose binding lectins, have different modes of action at the cellular levels, and thus generalizations should be treated with caution.
Sauvion
Nicolas
N
INRA-INSA de Lyon, UMR Biologie Fonctionnelle Insectes et Interactions, Bat. Louis-Pasteur, 20, ave. A. Einstein, 69621 Villeurbanne, France. sauvion@ensam.inra.fr
Nardon
Christiane
C
Febvay
Gérard
G
Gatehouse
Angharad M R
AM
Rahbé
Yvan
Y
D000818:Animals; D001042:Aphids / Q000033:anatomy & histology / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D003208:Concanavalin A / Q000378:metabolism / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D041981:Gastrointestinal Tract / Q000187:drug effects* / Q000378:metabolism; D007150:Immunohistochemistry; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D010572:Pest Control, Biological
D003208:Concanavalin A