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J. Biochem, 2003, Vol. 134, No. 1 143-150
© 2003 Japanese Biochemical Society

MOLECULAR BIOLOGY

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

Hideo Saito1,2, Kazuhisa Takeda1, Ken-ichi Yasumoto1, Haruo Ohtani+,3, Ken-ichi Watanabe1, Kazuhiro Takahashi1, Atsushi Fukuzaki2, Yoichi Arai2, Hiroaki Yamamoto4 and Shigeki Shibahara§,1

1 Department of Molecular Biology and Applied Physiology, 2 Department of Urology, and 3 Department of Pathology, Tohoku University School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575; and 4 Biological Institute, Graduate School of Life Sciences, Tohoku University, Aoba-ku, Sendai, Miyagi 980-8578

The gene coding for microphthalmia-associated transcription factor (Mitf) 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.

+ Present address: Research Division and Clinical Laboratory, Mito National Hospital, Mito 310-0035.

§ To whom correspondence should be addressed. E-mail: shibahar{at}mail.tains.tohoku.ac.jp


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