Uptake, Intracellular Distribution, and Stability of Oligodeoxynucleotide Phosphorothioate bySchistosoma mansoni

• DOI: 10.1089/ard.1995.5.123
• Publication Date: 2016-05-17T20:45:43Z
• Authors: LIANG-FENG TAO; KENNETH A. MARX; WARANYA WONGWIT; ZHIWEI JIANG; SUDHIR AGRAWAL; ROBERT M. COLEMAN
• Keywords: Male; 0303 health sciences; Base Sequence; Molecular Sequence Data; Schistosoma mansoni; Oligonucleotides, Antisense; Thionucleotides; Sulfur Radioisotopes; Antiviral Agents; Oligodeoxyribonucleotides, Antisense; Kinetics; 03 medical and health sciences; Drug Stability; Culture Techniques; Animals; Feasibility Studies; Female; Subcellular Fractions

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).