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BOR - Papers in Press, published online ahead of print September 15, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.032516
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Submitted June 16, 2004
Returned for revision June 29, 2004
Accepted September 3, 2004

Gamete Biology


Molecular Cloning of Rat Spetex-2 Family Genes Mapped on Chromosome 15p16, Encoding a 23-Kilodalton Protein Associated with the Plasma Membranes of Haploid Spermatids

Yuka Iwamoto , Takane Kaneko , Junko Ichinose , Takayuki Mori , Yosaburo Shibata , Kiyotaka Toshimori , and Hiroshi Iida *

* To whom correspondence should be addressed. E-mail: iidahiro{at}agr.kyushu-u.ac.jp.

Abstract
We used differential display in combination with complementary DNA cloning approach to isolate a novel rat gene designated as Spetex-2, which has an open reading frame of 582-length nucleotides encoding a protein of 194 amino acids. Spetex-2 mRNA was highly expressed in testis and spleen, and its expression in rat testis was developmentally up-regulated. In situ hybridization revealed that Spetex-2 mRNA was predominantly expressed in haploid spermatids of step 1~13 within the seminiferous epithelium. BLAST-search against rat genome data bases at NCBI revealed that Spetex-2 gene is composed of 4 exons and that it is mapped at least 18 loci in a cluster on rat chromosome 15p16, indicating that the genes occur as repeated tandem array over a long stretch of genomic DNA. By immunocytochemical analysis with confocal laser-scanning microscopy, Spetex-2 protein was detected as dot-like distribution on the cell periphery of haploid spermatids (step 1-13), but not observed in other spermatogenic cells. On the basis of these data, we hypothesize that Spetex-2 might be correlated with cell differentiation of spermatids in rat testis.

Key words: Testis • Gametogenesis • Sperm maturation • Spermatid • Spermatogenesis


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