Characterization of Pseudomonas aeruginosa adherence to mouse corneas in organ culture.
The present study was designed to obtain further information on the nature of the corneal macromolecule(s) to which Pseudomonas aeruginosa adheres and how adherence might be prevented. Scarified adult mouse corneas in organ culture were treated with trypsin or lipase to determine whether the receptor molecule(s) was protein or lipid in nature. Trypsin (20 micrograms/ml) treatment of the cornea for 5 min had no significant effect on bacterial adherence, and longer periods of enzyme exposure resulted in extensive surface cell lysis. In contrast, lipase treatment (50,000 U/ml) for 1 h caused little visible cell lysis and significantly reduced bacterial adherence. To test further the lipid nature of the receptor, a highly purified monosialoganglioside (GM1) preparation (500 micrograms/ml) was used to preincubate (1 h) the cornea prior to bacterial application, and this also inhibited bacterial adherence. Similar corneal treatment with gangliotetraosylceramide (asialo GM1) (500 micrograms/ml) had little effect on ocular bacterial binding. Premixing of the bacterial inoculum with GM1 prior to corneal application had no significant effect on inhibiting bacterial binding, but similarly premixing the bacterial inoculum with asialo GM1 transiently decreased adherence. Lastly, premixing of the bacterial inoculum or preincubation of corneas with fibronectin (500 micrograms/ml for 1 h) both decreased bacterial adherence. These findings provide evidence that the receptor-adhesin interactions of P. aeruginosa at the ocular surface in organ culture are complex, involve a glycolipid moiety, and may be blocked by a ganglioside containing at least one sialosyl residue or by fibronectin, which may bind to membrane-associated gangliosides.
Singh
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Department of Anatomy/Cell Biology, Wayne State University School of Medicine, Detroit, Michigan 48201.
Hazlett
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LD
Berk
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RS
D000818:Animals; D001422:Bacterial Adhesion; D003315:Cornea / Q000382:microbiology* / Q000473:pathology; D005260:Female; D005353:Fibronectins / Q000494:pharmacology; D005677:G(M1) Ganglioside / Q000494:pharmacology; D006028:Glycosphingolipids / Q000494:pharmacology; D008049:Lipase / Q000494:pharmacology; D051379:Mice; D008855:Microscopy, Electron, Scanning; D009924:Organ Culture Techniques; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000472:pathogenicity*; D014357:Trypsin / Q000494:pharmacology
D005353:Fibronectins; D006028:Glycosphingolipids; D005677:G(M1) Ganglioside; D008049:Lipase; D014357:Trypsin