Giardia lamblia rearranges F-actin and alpha-actinin in human colonic and duodenal monolayers and reduces transepithelial electrical resistance.
The mechanisms of epithelial injury in giardiasis remain unknown. The effects of live Giardia lamblia on cellular G-actin, F-actin, alpha-actinin, and electrical resistance of human intestinal epithelial monolayers were investigated using SCBN and Caco2 cell lines grown on chamber slides or Transwell filter membranes. In separate experiments, some monolayers were also exposed to sonicated trophozoites, some to supernatant from live G. lamblia cultures, and some with or without the Ca2+ channel blocker verapamil. After 2, 24, or 48 hr of coincubation with G. lamblia, monolayers were assessed for cytoskeletal arrangement under fluorescence and confocal laser microscopy, and transepithelial electrical resistance was measured. Exposure to live G. lamblia trophozoites induced localized condensation of F-actin and loss of perijunctional alpha-actinin while G-actin remained unchanged. Confocal laser microscopy indicated that F-actin rearrangement was not affected by verapamil and was localized within the terminal web area. Coincubation of monolayers with G. lamblia lysates or with spent medium alone similarly rearranged F-actin. Verapamil alone did not alter F-actin. Electrical resistance of SCBN and Caco2 monolayers exposed to G. lamblia was significantly decreased versus controls regardless of whether live or lysed trophozoite samples were used. The results indicate that G. lamblia-induced epithelial injury is associated with F-actin and alpha-actinin rearrangements in the terminal web area via mechanisms independent of extracellular Ca2+. These alterations are associated with reduced transepithelial electrical resistance and are due at least in part to trophozoite products.
Teoh
D A
DA
Department of Biological Sciences, University of Calgary, Alberta, Canada.
Kamieniecki
D
D
Pang
G
G
Buret
A G
AG
D000185:Actinin / Q000187:drug effects / Q000378:metabolism*; D000199:Actins / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D018938:Caco-2 Cells; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002460:Cell Line; D003106:Colon / Q000166:cytology / Q000187:drug effects / Q000469:parasitology* / Q000502:physiology; D004386:Duodenum / Q000166:cytology / Q000187:drug effects / Q000469:parasitology* / Q000502:physiology; D017097:Electric Impedance; D016829:Giardia lamblia / Q000502:physiology*; D006801:Humans; D007413:Intestinal Mucosa / Q000469:parasitology / Q000502:physiology*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D014700:Verapamil / Q000494:pharmacology
D000199:Actins; D002121:Calcium Channel Blockers; D000185:Actinin; D014700:Verapamil