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Flow cytometric analysis of the in situ hybridization of cyclooxygenase isoforms in mesangial cells treated with cyclosporine A
Author(s) -
de Hornedo J. Pérez,
Fernández M. Calvino,
de la Fuente G. de Arriba,
Fernández P. del Reino,
Cid T. Parra,
Martinez S. Benito
Publication year - 2006
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20230
Subject(s) - flow cytometry , in situ hybridization , cyclooxygenase , gene isoform , microbiology and biotechnology , biology , gene expression , messenger rna , chemistry , gene , biochemistry , enzyme
Abstract Background Cyclosporine A increases oxidative stress in kidney and we hypothesized that cyclooxygenase (COX) may be involved in this effect. Material and Methods Mesangial cells of Cyclosporine A‐treated (4, 7 or 10 days) rats were obtained to evaluate mRNA expression of COX‐isoforms (COX‐1, constitutive and COX‐2, inducible) by “in situ” hybridization. Probes were labelled using “Gene Image Random Prime Labelling Protocol” and COX expression was measured by flow cytometry. Results and Discussion “In situ” hybridization by flow cytometry is an useful method to detect mRNA. We observed an increased COX‐2 expression in a time‐dependent manner in parallel with Reactive Oxygen Species synthesis. COX‐1 expression increased only at 10 days. © 2006 International Society for Analytical Cytology