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Internalization of oligodeoxynucleotide antisense to type‐1 plasminogen activator inhibitor mRNA in endothelial cells: A three‐dimensional reconstruction by confocal microscopy
Author(s) -
Wyroba Elzbieta,
Pawlowska Zofia,
Kobylanska Anna,
Pluskota Elzbieta,
Maszewska Maria,
Stec Wojciech J,
Cierniewski Czeslaw S
Publication year - 1996
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1111/j.1768-322x.1996.tb00964.x
Subject(s) - biology , cytoplasm , oligonucleotide , microbiology and biotechnology , internalization , phosphodiester bond , plasminogen activator , confocal microscopy , incubation , cell , biochemistry , rna , dna , endocrinology , gene
Summary— A three‐dimensional reconstruction analysis of localization of phosphodiester and phosphorothioate oligonucleotide antisense to type‐I plasminogen activator inhibitor (PAI‐1) mRNA within endothelial cells is described. When EA.hy 926 cells were incubated with fluorescently labelled phosphodiester (PO‐16) or phosphorothioate (PS‐16) oligonucleotides at low, not cytotoxical concentrations, the relative brightness composition of the images of the particular samples was much higher for PS‐16 than PO‐16 and dependent upon the extracellular concentration and the incubation time. The 3‐D reconstructions based on the series of optical sections of the samples, spaced every 1.5gμm, showed the punctuate accumulation of the oligonucleotides and a striking difference in a spatial distribution between PO‐16 and PS‐16 within the cytoplasm. Even after 24 h incubation of endothelial cells with 2.5 μM of PO‐16 and PS‐16 oligonucleotides, there was a predominant oligonucleotide localization within the cytoplasm and only traces of oligonucleotides could be seen in the cell nucleus and/or perinuclear organelles.