Uptake, intracellular distribution, and stability of oligodeoxynucleotide phosphorothioate by Schistosoma mansoni.
The in vitro uptake, cellular distribution, efflux, stability, and toxicity levels of an oligodeoxynucleotide phosphorothioate (PS-oligonucleotide) have been studied in mature Schistosoma mansoni worms. The intracellular accumulation of 35S-labeled PS-oligonucleotide occurred roughly in proportion to the worm body mass over a wide concentration range, whether the worms were exposed singly or in mating pairs. Cellular uptake was dependent on the extracellular concentration. A minor fraction (13%) of the PS-oligonucleotide taken up by the worm accumulated in the surface tegumental coat. Most of the PS-oligonucleotide taken up localized in the cytosol (54%) and the nuclei-enriched (33%) fractions. In a time course study on adult worms in culture, oligonucleotide uptake was observed within the first 2 h and peaked at about 36 h. A decrease in the intracellular concentration of the PS-oligonucleotide was observed by 42 h. Analysis of the extracted oligonucleotides showed that PS-oligonucleotide was digested slowly. Efflux of the oligonucleotide was time and temperature dependent. Significant toxicity to the cultured worms did not occur until the PS-oligonucleotide concentration was over 8 mg/ml (1 mM).
Tao
L F
LF
Center for Tropical Diseases, University of Massachusetts-Lowell 01854, USA.
Marx
K A
KA
Wongwit
W
W
Jiang
Z
Z
Agrawal
S
S
Coleman
R M
RM
D000818:Animals; D000998:Antiviral Agents; D001483:Base Sequence; D046508:Culture Techniques; D004355:Drug Stability; D005240:Feasibility Studies; D005260:Female; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D020319:Oligodeoxyribonucleotides, Antisense; D016376:Oligonucleotides, Antisense / Q000378:metabolism* / Q000633:toxicity; D012550:Schistosoma mansoni / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D013462:Sulfur Radioisotopes; D013873:Thionucleotides / Q000378:metabolism* / Q000633:toxicity
D000998:Antiviral Agents; D020319:Oligodeoxyribonucleotides, Antisense; D016376:Oligonucleotides, Antisense; D013462:Sulfur Radioisotopes; D013873:Thionucleotides; C079996:GEM91