Purpose. The biological effects of inducible nitric oxide synthase (iNOS) in experimental meitis are not clear. In order to study which cells contribute to intraocular NO synthesis in inflammatory conditions, we have studied the distribution of iNOS+ cells in iris whole mounts of Lewis rats with endoloxin-induced uveitis (EIU). Methods. Iris whole mounts were prepared from rats 24 h after endotoxin injection. The whole mounts \\ere processed for overnight incubations, first in a solution of 0.1 % triton PBS followed by a 1:100 dilution of a commercially available mouse anti-rat iNOS monoclonal antibody (Transduction Laboratories SC) and finally in a 1:200 dilution of FI rC-conjugatcd anti-mouse IgG antibody (Immunoresearch Laboratories Inc.). Results. In rats with FTU. large numbers of 1NOS+ cells could be detected in the iris whole mounts. The iNOS + cells were localized mainly within ihe iris vessels, mostly showing an ouiid or round appearance. Moreover, endothelial cells were always negative, indicating that no cross reaction with endothelial constitutive NOS occurs. Further preliminary observations indicated that the relative number of extravascularly detected iNOSi cells is found to increase with severity of the disease. Conclusions. Immunohistochemical analysis for iNOS+ cells in iris whole mounts provides a tool lor the study of iNOS expression in EIU. It may allow the elucidation of the protective or negative role of iNOS in uveitis under different experimental conditions.
|Investigative Ophthalmology and Visual Science
|Published - 1 Dec 1997