Visualization of Mitochondrial and Apicoplast Nucleoids in the Human Malaria Parasite Plasmodium falciparum by SYBR Green I and PicoGreen Staining

• Maeda-Sano Katsura

  Division of Biology, Faculty of Science, Ochanomizu University

• Sato Shigeharu

  Division of Parasitology, MRC National Institute for Medical Research

• Ueda Takashi

  Department of Biological Sciences, Graduate School of Science, The University of Tokyo

• Yui Ryoko

  Division of Biological Science, Graduate School of Science, Nagoya University

• Ito Kie

  Division of Biology, Faculty of Science, Ochanomizu University Division of Biological Science, Graduate School of Science, Nagoya University Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo

• Hata Masayuki

  Department of Biomedical Chemistry, Graduate School of Medicine, The University of Tokyo

• Nakano Akihiko

  Department of Biological Sciences, Graduate School of Science, The University of Tokyo RIKEN Advanced Science Institute

• Kita Kiyoshi

  Department of Biomedical Chemistry, Graduate School of Medicine, The University of Tokyo

• Murakami-Murofushi Kimiko

  Division of Biology, Faculty of Science, Ochanomizu University

• Sasaki Narie

  Division of Biological Science, Graduate School of Science, Nagoya University

Abstract

Organellar DNA in mitochondria and plastids are organized with proteins into a compact structure known as the nucleoid. As the nucleoid is supposed to be the unit of inheritance for the organellar genome, it is important to understand its cytological behavior. Like plants, malaria parasites carry two organelles, the mitochondrion and the apicoplast–a non-photosynthetic plastid. However, probably because of the small size of the genome in each, visualizing the nucleoid in the Plasmodium organelles by regularly-used fluorescent dye such as DAPI, has been difficult. Here, we developed new, effective methods to visualize the organellar nucleoid in the human malaria parasite Plasmodium falciparum. With our methods using SYBR Green I or PicoGreen, nucleoids were observed in ring-stage parasites. Analyzing transfectant parasites carrying a DsRed-labelled organelle, we concluded that the parasite's mitochondrion has 1 nucleoid which is visualized with our methods. The parasite has a second nucleoid in the apicoplast, but higher concentration of the dye was required to visualize it. Our new methods would be useful for further cytological analysis of the nucleoids in the mitochondrion and the apicoplast of the malaria parasite.

Journal

• CYTOLOGIA

  CYTOLOGIA 74 (4), 449-455, 2009

  Japan Mendel Society, International Society of Cytology

Keywords

• Mitochondrial DNA (mtDNA)
• Apicoplast DNA (apDNA)
• Nucleoid
• SYBR Green I
• PicoGreen
• <i>Plasmodium falciparum</i>

Details

• CRID: 1390001204108736768
• NII Article ID: 10026186344
• NII Book ID: AA00622764
• DOI: 10.1508/cytologia.74.449
• ISSN: 13487019
• ISSN: 00114545
• Web Site: http://www.jstage.jst.go.jp/article/cytologia/74/4/74_4_449/_pdf
• Text Lang: en
• Data Source:
  • JaLC
  • Crossref
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed