Tissue distribution of a menthyl-conjugated oligodeoxyribonucleotide antisense to PAI-1 mRNA.
The inhibitory effect of numerous analogues of PO-16, an hexadecadeoxyribonucleotide antisense to sequences -22 to -17 of PAI-1 mRNA coding for a fragment of the signal peptide, on the expression of PAI-1 in endothelial cells, and physiological consequences of the subsequently reduced PAI-1 activity tested in vitro and in vivo, were described in our previous studies. Of particular interest was PO-16 5'-O-conjugated with menthyl phosphorothioate (MPO-16R). In this work, tissue localisation of MPO-16R labelled with [(35)S] phosphorothioate at the 3'-end, was determined. [(35)S]MPO-16R and control [(35)S]MPO-16R-SENSE oligonucleotides were administered intravenously into 22 rats and organ distribution of the labelled bioconjugates was assessed after 24 and 48 h. For this purpose, tissue sections were subjected to autoradiography, and quantitated by liquid scintillation after solubilisation. Overall clearance of radioactivity was already seen after 24 h, with the radioactivity recovered mainly in the kidney and liver. A smaller fraction of radioactivity was also retained in the spleen and heart. The kidney concentration of the labelled probe was higher than that of liver by 50%. The distribution of PAI-1 mRNA in untreated rat kidney, liver, spleen and heart established by two independent techniques: Ribonuclease Protection Assay and Real-Time PCR, shows the same pattern as that observed for [(35)S]MPO-16R antisense.
Szemraj
Janusz
J
Department of Medical Biochemistry, Medical University in Łódź, Poland.
Al-Nedawi
Khalid N I
KN
Chabielska
Ewa
E
Buczko
Wlodzimierz
W
Pawlowska
Zofia
Z
D000818:Animals; D001483:Base Sequence; D008297:Male; D008745:Methylation; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D015345:Oligonucleotide Probes / Q000097:blood; D016376:Oligonucleotides, Antisense / Q000493:pharmacokinetics*; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics*; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar
D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D016376:Oligonucleotides, Antisense; D017395:Plasminogen Activator Inhibitor 1; D021382:Protein Sorting Signals; D012333:RNA, Messenger