Intravital Microscopy of the Murine Urinary Bladder Microcirculation
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OBJECTIVE: To establish an in vivo mouse model of the urinary bladder microcirculation, and characterize the molecular mechanisms of endotoxin-induced leukocyte recruitment. METHODS: The murine model was adapted from a technique previously reported for the rat. Mouse bladder microcirculation was observed using intravital microscopy, four hours after intravesical challenge with lipopolysaccharide (LPS) and leukocyte-endothelial interactions were examined. Molecular mechanisms of leukocyte recruitment were identified using antibodies to adhesion molecules and chemokines. RESULTS: LPS from Escherichia coli administered intravesically resulted in a significant increase in leukocyte adhesion and rolling at four hours post stimulation. LPS from Pseudomonas aeruginosa administered at similar doses resulted in a significant, but lower increase in leukocyte adhesion after four hours compared with E. coli LPS. Leukocyte adhesion within the bladder microcirculation was dependent on α(4) -integrins and ICAM-1, whereas leukocyte rolling was P-selectin dependent, but α(4) -integrin independent. Blockade of MIP-2 and KC did not alter leukocyte-endothelial interactions. The bladder endothelium expressed P-selectin, ICAM-1, VCAM-1, MIP-2, and MCP-1. Only VCAM-1 endothelial expression was significantly increased after LPS stimulation. CONCLUSION: The mouse model of the urinary bladder microcirculation is suitable for the study of inflammatory responses during urinary tract infection (UTI) in vivo.
