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ARTICLE |
Cell Type-specific Localization of Sphingosine Kinase 1a in Human Tissues
Takashi Muratea, Yoshiko Bannob, Keiko T-Koizumic, Kazuko Watanabed, Naoyoshi Morid, Atsushi Wadae, Yasuyuki Igarashie, Akira Takagia, Tetsuhito Kojimaa, Haruhiko Asanof, Yukihiro Akaog, Shonen Yoshidac, Hidehiko Saitof, and Yoshinori Nozawaga Nagoya University School of Health Science, Nagoya, Japan
b The First Department of Biochemistry, Gifu University School of Medicine, Gifu, Japan
c The Disease Mechanism and Control, Nagoya University School of Medicine, Nagoya, Japan
d The First Department of Pathology, Nagoya University School of Medicine, Nagoya, Japan
e Department of Biomembrane and Biofunctional Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University, Hokkaido, Japan
f The First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan
g Gifu International Institute of Biotechnology, Gifu, Japan
Correspondence to: Takashi Murate, Nagoya U. School of Health Science, Daiko-minami, 1-1-20, Higashi-ku, Nagoya 461-8673, Japan. E-mail: murate@met.nagoya-u.ac.jp
Cell type-specific localization of sphingosine kinase 1a (SPHK1a) in tissues was analyzed with a rabbit polyclonal antibody against the 16 C-terminal amino acids derived from the recently reported mouse cDNA sequence of SPHK1a. This antibody (anti-SPHK1a antibody) can react specifically with SPHK1a of mouse, rat, and human tissues. Utilizing its crossreactivity to human SPHK1a, the cell-specific localization of SPHK1a in human tissues was histochemically examined. Strong positive staining for SPHK1a was observed in the white matter in the cerebrum and cerebellum, the red nucleus and cerebral peduncle in the midbrain, the uriniferous tubules in the kidney, the endothelial cells in vessels of various organs, and in megakaryocytes and platelets. The lining cells of sinusoids in the liver and splenic cords in the spleen showed moderate staining. Columnar epithelia in the intestine and Leydig's cells in the testis showed weak staining patterns. In addition, TPA-treated HEL cells, a human leukemia cell line, showed a megakaryocytic phenotype accompanied with increases in immunostaining of both SPHK1a and SPHK enzyme activity, suggesting that SPHK1a may be a novel marker of megakaryocytic differentiation and that this antibody is also useful for in vitro study of differentiation models.
(J Histochem Cytochem 49:845–855, 2001)
Key Words: sphingosine-1-phosphate, sphingosine kinase, polyclonal antibody, C-terminal amino acids, immunohistochemistry, tissue distribution
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