Bioimaging Analysis of Cellular Uptake of Cholesterol-Conjugated Oligonucleotides Using a Real-Time Confocal Laser Microscope

• Hanaki Kenichi

  The Chemo-Sero-Therapeutic Research Institute Department of Bacteriology, Faculty of Medicine, University of Tokyo

• Ueno Tetsuro

  The Chemo-Sero-Therapeutic Research Institute

• Nakatake Hiroshi

  The Chemo-Sero-Therapeutic Research Institute

• Nozaki Chikateru

  The Chemo-Sero-Therapeutic Research Institute

• Taniguchi Takahide

  Veterinary Microbiology, Tokyo University of Agriculture and Technology

• Honda Eiichi

  Veterinary Microbiology, Tokyo University of Agriculture and Technology

• Hayashi Masanobu

  Department of Veterinary Radiology, Rakuno Gakuen University

• Yamamoto Kenji

  Laboratory of Clinical Microbiology and Immunology, Bun'in Hospital of University of Tokyo

Abstract

The uptake of 3'-cholesterol conjugated phosphorothioate oligonucleotides with 5'-fluorescein (ChPSf-ODN) into rat immortalized hepatocytes was visualized with a real-time confocal laser microscope. The images showed that the uptake of ChPSf-ODN in the cytosol had started within the first 5 minutes, and it was not until a 40 minute-incubation that the endosomal and/or lysosomal accumulation appeared. Histogram analysis of fluorescence intensity indicated that cytosolic uptake of ChPSf-ODN was five times as great as that of phosphorothioate oligonucleotides with 5'-fluorescein (PSf-ODN). However, nuclear uptake of ChPSf-ODN was only twice as great as that of PSf-ODN. The mechanisms of the different amounts of ChPSf-ODN distribution in the subcellular compartments are discussed. This study shows that the visual data derived from time-lapse confocal laser microscopy are helpful in understanding the cellular uptake mechanism of fluorescein labeled ODN.

Journal

• bioimages

  bioimages 5 (4), 127-132, 1997

  Bioimaging Society

Keywords

• oligonucleotide
• cholesterol conjugation
• cellular uptake
• time-lapse confocal laser microscopy

Details

• CRID: 1390007857409391872
• NII Article ID: 10002038305
• NII Book ID: AA11084187
• DOI: 10.11169/bioimages.5.127
• ISSN: 09192719
• Text Lang: en
• Data Source:
  • JaLC
  • CiNii Articles
• Abstract License Flag: Disallowed