Identifier

etd-11092005-125857

Degree

Master of Science (MS)

Department

Biomedical and Veterinary Medical Sciences - Veterinary Clinical Sciences

Document Type

Thesis

Abstract

Urokinase (uPA) and urokinase receptor (uPAR) are present in the urinary tract of people, mice, rats and cows. The presence of urokinase and urokinase receptor in the urinary tract of dogs has not been documented. Documenting the presence of uPA and uPAR in the urinary tract of healthy dogs will help determine the role these proteins play in pathology of the urinary tract in further research. Urine was collected from fifty-four healthy dogs. A fluorescence assay was developed using the fluorogenic substrate Z-Gly-Gly-Arg-AMC, HCl. This assay documented presence of active uPA-like protein in concentrated urine. A direct sandwich ELISA assay was developed using a mouse anti-mouse primary antibody and a biotin labeled rabbit anti-human secondary antibody. This ELISA documented presence of active and inactive uPA-like protein and measured concentration of uPA-like protein in concentrated urine. Tissues from the urinary tract of eleven normal dogs (four intact females, five intact males and two castrated males) were collected from kidney, ureter, urinary bladder, urethra, prostate and testicle. SDS-PAGE/Western blot analysis using mouse anti-mouse, mouse anti-human and rabbit anti-human uPA antibodies and a mouse anti-human uPAR antibody were used to identify uPA-like and uPAR-like protein on the basis of molecular weight and antigen-antibody reactions. Immunoprecipitation was employed to enhance identification uPA-like and uPAR-like protein using mouse anti-mouse, mouse anti-human and rabbit anti-human uPA antibodies and a mouse anti-human uPAR antibody. Immunohistochemistry was utilized to identify staining in specific cell types within the urinary tract. The uPA antibodies used were directed against human and mouse urokinase. The uPAR antibody used was directed against human urokinase receptor. A urokinase-like protein is present in urine from healthy dogs in small but measurable quantities. Urokinase-like and urokinase receptor-like protein are present in the urinary tract of the healthy dog. This distribution is similar to that identified in people. The role of urokinase-like and urokinase receptor-like protein in the urinary tract of the normal dog is yet to be defined.

Date

2005

Document Availability at the Time of Submission

Release the entire work immediately for access worldwide.

Committee Chair

Giselle Hosgood

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