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Characterization of the differential expression of uncoupling protein 2 and ROS production in differentiated mouse macrophage-cells (Mm1) and the progenitor cells (M1)

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Abstract

The expression status of mitochondrial uncoupling protein 2 (UCP2) was investigated in undifferentiated mouse myeloid leukemia (M1) and its differentiated macrophage-like cells (Mm1). Mm1 cells have a high ability of phagocytosis along with significantly high levels of reactive oxygen species (ROS) production, UCP2 protein and manganese superoxide dismutase (Mn-SOD), in contrast to undifferentiated leukemia cells (M1). Mm1 cells expressed 10-fold more UCP2 protein compared with undifferentiated M1 cells, although the UCP2 mRNA levels in both cell types were similar. The higher expression of UCP2 in the Mm1 cells suggests a regulatory role of UCP2 in the ROS production. Furthermore, the transfection of UCP2-GFP-expression vector in Mm1 cells dissipated the mitochondrial membrane potential and reduced ROS production, which was shown by their direct visualization using MitoTracker Red CM-H2Xros. The macrophage gp91phox protein, a membrane catalytic component of the NADPH oxidase complex, was at a similar level in both of UCP2-GFP expressed and non-expressed Mm1 cells. These results suggest that the UCP2 protein of the undifferentiated cell is regulated at a quite low level and the higher UCP2 protein of the differentiated macrophages involves with the regulation of ROS production.

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Abbreviations

UCP2:

uncoupling protein 2

ROS:

reactive oxygen species

DHR123:

Dihydrorhodamine 123

NBT:

Nitro Blue Tetrazolium

ORF:

open reading frame

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Author notes

  1. Koji Nishio

    Present address: Department of Biochemistry, Aichi Medical College, Nagakute, Aichi, Japan, 480-1195

Authors and Affiliations

  1. Department of Anatomy and Cell Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, 466-8550, Showa-ku, Nagoya, Aichi, Japan

    Koji Nishio & Shanlou Qiao

  2. Department of Molecular Genetics, National Institute for Longevity Sciences, 36-3 Gengo, Aichi, Obu, Japan, 474-8522

    Hitoshi Yamashita

Authors
  1. Koji Nishio
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  2. Shanlou Qiao
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  3. Hitoshi Yamashita
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Correspondence to Koji Nishio.

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Nishio, K., Qiao, S. & Yamashita, H. Characterization of the differential expression of uncoupling protein 2 and ROS production in differentiated mouse macrophage-cells (Mm1) and the progenitor cells (M1). J Mol Hist 36, 35–44 (2005). https://doi.org/10.1007/s10735-004-2915-x

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  • DOI: https://doi.org/10.1007/s10735-004-2915-x

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