Germ Cell-Specific Expression of Microphthalmia-Associated Transcription Factor mRNA in Mouse Testis

  • Saito Hideo
    Department of Molecular Biology and Applied Physiology Department of Urology, Tohoku University School of Medicine
  • Takeda Kazuhisa
    Department of Molecular Biology and Applied Physiology
  • Yasumoto Ken-ichi
    Department of Molecular Biology and Applied Physiology
  • Ohtani Haruo
    Department of Pathology, Tohoku University School of Medicine Research Division and Clinical Laboratory, Mito National Hospital
  • Watanabe Ken-ichi
    Department of Molecular Biology and Applied Physiology
  • Takahashi Kazuhiro
    Department of Molecular Biology and Applied Physiology
  • Fukuzaki Atsushi
    Department of Urology, Tohoku University School of Medicine
  • Arai Yoichi
    Department of Urology, Tohoku University School of Medicine
  • Yamamoto Hiroaki
    Biological Institute, Graduate School of Life Sciences, Tohoku University
  • Shibahara Shigeki
    Department of Molecular Biology and Applied Physiology

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The gene coding for microphthalmia-associated transcription factor (Mitt) contains many promoters that could generate multiple Mitf isoforms with distinct amino-termini, such as ubiquitously expressed Mitf-A and Mitf-H. To gain further insight into Mitf isoform multiplicity and the regulation of the promoter usage of the Mitf gene, we have analyzed the function of the amino-terminal domains of Mitf isoforms and the expression of Mitf mRNA in mouse postnatal testis, which is characterized by spermatogenesis and by a cool temperature because of its unique location. Here we show that the amino-terminal domain of Mitf-A possesses a transactivation activity, as judged by yeast expression analysis. We also show the expression of Mitf-A and Mitf-D mRNAs in testis by PCR-based methods. Moreover, in situ hybridization analysis revealed that an Mitf mRNA, probably representing Mitf-A and/or Mitf-D, is expressed in germ cells, including spermatogonia, spermatocytes that undergo meiosis, and round spermatids with the haploid genome, but is undetectable in elongated spermatids with remodeled and condensed chromatin. Notably, Mitf mRNA is undetectable in somatic Leydig cells and peritubular cells. Therefore, multiple promoters may direct differential expression of the Mitf gene in the testis and contribute to functional diversity of Mitf isoforms.

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