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Complementary DNA Cloning and Characterization of Rat spergen-2, a Spermatogenic Cell-Specific Gene 2 Encoding a 56-Kilodalton Nuclear Protein Bearing Ankyrin Repeat Motifs¹

Takayuki Mri³

Laboratory of Zoology,³ Graduate School of Agriculture, Kyushu University, Higashiku Hakozaki 6-10-1, Fukuoka 812-8581, Japan Department of Anatomy,⁴ Miyazaki Medical College, Miyazaki 889-1692, Japan Department of Developmental Anatomy,⁵ Graduate School of Medical Science, Kyushu University, 3-1-1 Maidashi, Fukuoka 812-8582, Japan

Differential display in combination with a cDNA cloning approach were used to isolate a novel gene, spergen-2, which has an open reading frame of 1500 nucleotides and encodes a protein of 500 amino acids that contains ankyrin repeat motifs and a putative nuclear localization signal. Expression of spergen-2 is developmentally upregulated in testis. In situ hybridization revealed that spergen-2 mRNA is expressed in spermatocytes and round spermatids (steps 1–6). Immunohistochemical analysis with confocal laser-scanning microscopy demonstrated that spergen-2 protein is predominantly expressed in nuclei of late spermatocytes (stages IX–XIV) and spermatids (steps 1–11), indicating the restricted expression of spergen-2 during spermatogenesis. In nucleoplasm of spermatogenic cell nuclei, spergen-2 tends to localize in the interchromosome space with relatively low DNA density. These findings indicate a potential role of spergen-2 in spermatogenesis, especially in cell differentiation from late pachytene spermatocytes to spermatids or in early spermatid differentiation.

¹ This work was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science, Takeda Science Foundation, and the Narishige Zoological Science Award.

² Correspondence. FAX: 92 642 2804; iidahiro{at}agr.kyushu-u.ac.jp

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