Iida Hiroshi, Ichinose Junko, Kaneko Takane, Mōri Takayuki, Shibata Yosaburo
Laboratory of Zoology, Graduate School of Agriculture, Kyushu University, Higashiku Hakozaki, Fukuoka, Japan.
Mol Reprod Dev. 2004 Aug;68(4):385-93. doi: 10.1002/mrd.20101.
We used differential display in combination with complementary DNA (cDNA) cloning approach to isolate a novel rat gene designated as spetex-1, which had an open reading frame of 1,668-length nucleotides encoding a protein of 556 amino acids. Spetex-1 mRNA was highly expressed in testis, and weekly expressed in lung, intestine, and spleen. Spetex-1 expression in the rat testes was detected first at 3 weeks in postnatal development and continued to be detected up to adulthood. A search in the databases showed that the amino acid sequence of spetex-1 was 82% identical to that of its mouse homologue found in the databases. Both rat spetex-1 and the mouse homologue contained Ser-X (X = His, Arg, or Asn) repeats in the middle portion of the proteins. In situ hybridization revealed that spetex-1 mRNA was expressed in haploid spermatids of step 7-18 within the seminiferous epithelium. Immunohistochemical analysis with confocal laser-scanning microscopy demonstrated that spetex-1 protein was not expressed in spermatogonia, spermatocytes, and round spermatids in adult rat testis, but was specifically detected in the residual cytoplasm of elongate spermatids of step 15-18 as well as in residual bodies engulfed by Sertoli cells. We interpreted these data as a potential role of spetex-1 in spermatogenesis, especially in cell differentiation from late elongate spermatids to mature spermatozoa.
我们采用差异显示技术结合互补DNA(cDNA)克隆方法,分离出一个新的大鼠基因,命名为spetex-1,其开放阅读框为1668个核苷酸长度,编码一个由556个氨基酸组成的蛋白质。Spetex-1 mRNA在睾丸中高度表达,在肺、肠和脾脏中低水平表达。在大鼠睾丸中,Spetex-1的表达在出生后3周首次被检测到,并持续到成年期。数据库检索显示,spetex-1的氨基酸序列与其在数据库中找到的小鼠同源物的氨基酸序列有82%的同一性。大鼠spetex-1和小鼠同源物在蛋白质的中间部分都含有Ser-X(X = His、Arg或Asn)重复序列。原位杂交显示,spetex-1 mRNA在生精上皮内第7-18步的单倍体精子细胞中表达。共聚焦激光扫描显微镜免疫组织化学分析表明,spetex-1蛋白在成年大鼠睾丸的精原细胞、精母细胞和圆形精子细胞中不表达,但在第15-18步伸长精子细胞的残余细胞质以及被支持细胞吞噬的残余小体中被特异性检测到。我们将这些数据解释为spetex-1在精子发生中,特别是在从晚期伸长精子细胞到成熟精子的细胞分化中具有潜在作用。