Corneal surface glycosylation is modulated by IL-1R and Pseudomonas aeruginosa challenge but is insufficient for inhibiting bacterial binding

Amber L Jolly; Paresh Agarwal; Matteo M E Metruccio; David R Spiciarich; David J Evans; Carolyn R Bertozzi; Suzanne M J Fleiszig

doi:10.1096/fj.201601198R

Corneal surface glycosylation is modulated by IL-1R and Pseudomonas aeruginosa challenge but is insufficient for inhibiting bacterial binding

FASEB J. 2017 Jun;31(6):2393-2404. doi: 10.1096/fj.201601198R. Epub 2017 Feb 21.

Authors

Amber L Jolly¹, Paresh Agarwal², Matteo M E Metruccio¹, David R Spiciarich², David J Evans^{1

3}, Carolyn R Bertozzi^{2

4

5}, Suzanne M J Fleiszig^{6

7

8

9}

Affiliations

¹ School of Optometry, University of California, Berkeley, Berkeley, California, USA.
² College of Chemistry, University of California, Berkeley, Berkeley, California, USA.
³ College of Pharmacy, Touro University California, Vallejo, California, USA.
⁴ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.
⁵ Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
⁶ School of Optometry, University of California, Berkeley, Berkeley, California, USA; fleiszig@berkeley.edu.
⁷ Graduate Division of Vision Sciences, University of California, Berkeley, Berkeley, California, USA.
⁸ Graduate Division of Microbiology, University of California, Berkeley, Berkeley, California, USA.
⁹ Graduate Division of Infectious Diseases and Immunity, University of California, Berkeley, Berkeley, California, USA.

Abstract

Cell surface glycosylation is thought to be involved in barrier function against microbes at mucosal surfaces. Previously we showed that the epithelium of healthy mouse corneas becomes vulnerable to Pseudomonas aeruginosa adhesion if it lacks the innate defense protein MyD88 (myeloid differentiation primary response gene 88), or after superficial injury by blotting with tissue paper. Here we explored their effect on corneal surface glycosylation using a metabolic label, tetra-acetylated N-azidoacetylgalactosamine (Ac₄GalNAz). Ac₄GalNAz treatment labeled the surface of healthy mouse corneas, leaving most cells viable, and bacteria preferentially associated with GalNAz-labeled regions. Surprisingly, corneas from MyD88^-/- mice displayed similar GalNAz labeling to wild-type corneas, but labeling was reduced and patchy on IL-1 receptor (IL-1R)-knockout mouse corneas (P < 0.05, ANOVA). Tissue paper blotting removed GalNAz-labeled surface cells, causing DAPI labeling (permeabilization) of underlying cells. MS of material collected on the tissue paper blots revealed 67 GalNAz-labeled proteins, including intracellular proteins. These data show that the normal distribution of surface glycosylation requires IL-1R, but not MyD88, and is not sufficient to prevent bacterial binding. They also suggest increased P. aeruginosa adhesion to MyD88^-/- and blotted corneas is not due to reduction in total surface glycosylation, and for tissue paper blotting is likely due to cell permeabilization.-Jolly, A. L., Agarwal, P., Metruccio, M. M. E., Spiciarich, D. R., Evans, D. J., Bertozzi, C. R., Fleiszig, S. M. J. Corneal surface glycosylation is modulated by IL-1R and Pseudomonas aeruginosa challenge but is insufficient for inhibiting bacterial binding.

Keywords: GalNAz metabolic label; Muc4; MyD88; imaging; mass spectrometry.

MeSH terms

Animals
Bacterial Adhesion
Cornea / microbiology*
Cornea / physiology*
Female
Fibrin Tissue Adhesive
Gene Expression Regulation / physiology
Glycoproteins
Glycosylation
Male
Mice
Mice, Inbred C57BL
Mice, Knockout
Myeloid Differentiation Factor 88 / genetics
Myeloid Differentiation Factor 88 / metabolism*
Pseudomonas aeruginosa
Receptors, Interleukin-1 / genetics
Receptors, Interleukin-1 / metabolism*

Substances

Fibrin Tissue Adhesive
Glycoproteins
Myd88 protein, mouse
Myeloid Differentiation Factor 88
Receptors, Interleukin-1

Abstract

MeSH terms

Substances

Grants and funding