Murine ocular heparanase expression before and during infection with Pseudomonas aeruginosa.
OBJECTIVE
To demonstrate the constitutive expression and regulation of heparanase (heparan sulfate endoglycosidase) in the normal mouse eye and in mice intracorneally infected with Pseudomonas aeruginosa.


METHODS
Naïve (unimmunized) and immunized C57BL/6J mice were infected with P. aeruginosa, and corneal heparanase gene and protein expression were detected by semiquantitative RT-PCR and immunoblot analysis. Immunohistochemistry was also applied to characterize corneal heparanase in naïve mice.


RESULTS
Heparanase mRNA and protein expression were detected in uninfected corneas of C57BL/6J mice. Immunohistochemical studies indicated heparanase protein expression was primarily in the corneal epithelium before corneal infection and was also in the corneal stroma after infection. Immunohistochemical studies of uninfected and infected whole eyes of naïve mice indicated heparanase protein expression in most layers of the retina, but the expression did not appear to be upregulated during corneal infection. Staining was most intense in the inner photoreceptor layer of the retina.


CONCLUSIONS
Heparanase was constitutively expressed in both the corneal epithelium and several retinal layers before intracorneal infection with P. aeruginosa. Temporal upregulation of corneal heparanase protein expression was detected in naïve mice during infection, most likely due to heparanase positive infiltrating cells, but the protein was not upregulated in corneas from immunized mice because they had a lower inflammatory response, associated with the restoration of corneal clarity. There did not appear to be temporal upregulation of heparanase expression in the retina of infected mice, as determined by immunohistochemistry.
Berk
Richard S
RS
Department of Immunology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA. rberk@med.wayne.edu
Dong
Zhong
Z
Alousi
Sarah
S
Kosir
Mary Ann
MA
Wang
Yuying
Y
Vlodavsky
Israel
I
D000818:Animals; D003315:Cornea / Q000201:enzymology*; D003320:Corneal Ulcer / Q000201:enzymology* / Q000382:microbiology; D015818:Eye Infections, Bacterial / Q000201:enzymology* / Q000382:microbiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D011552:Pseudomonas Infections / Q000201:enzymology* / Q000382:microbiology; D012333:RNA, Messenger / Q000378:metabolism; D012160:Retina / Q000201:enzymology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D000099175:Heparanase
D005966:Glucuronidase; D012333:RNA, Messenger; D000099175:Heparanase