Purpose. To examine whether messenger RNA for basic fibroblast growth factor (bFGF) could be visualized by a fluorescent probe in living trabecular cells. Methods. A 18-nucleotide-long antisense or sense sequence for human bFGF was sandwiched between two complementary 5-nucleotide-long arm sequences. A fluorophore, 5-(2′-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS), was joined to the 5′-terminal phosphate, while 4-(4′-dimethylaminophenylazo)benzoic acid, quencher for EDANS, was joined to the 3′-terminal hydroxyl group. The probe emitted blue fluorescence only upon hybridization with the complementary 18-nucleotide sequence when stimulated by ultraviolet light. The antisense or sense probe carried with liposome was transfeeted into human trabecular cells which were derived from trabecular tissues excised during trabeculectomy and cultured in a glass-bottom culture dish. The culture dish was placed on the stage of an inverted microscope and fluorescence was observed with a filter under ultraviolet light. Results. CelIs transfeeted with the antisense probe showed blue fluorescence under ultraviolet 1ight, while cells with the sense probe did not. Conclusions. Messenger RNA for bFGF could be visualized in living trabecular cells. The present study opens a way to measure the changing level of specific messenger RNA in living cells.
|Journal||Investigative Ophthalmology and Visual Science|
|Publication status||Published - Dec 1 1997|
ASJC Scopus subject areas
- Sensory Systems
- Cellular and Molecular Neuroscience