Ina Sawako, Tsunekawa Naoki, Nakamura Aya, Noce Toshiaki
Laboratory of Reproductive Biology, Mitsubishi Kasei Institute of Life Sciences, 11 Minami-Ooya, Machida-shi, 194-8511 Tokyo, Japan.
Gene Expr Patterns. 2003 Oct;3(5):635-8. doi: 10.1016/s1567-133x(03)00111-x.
As an attempt to investigate the function of the mouse Vasa-homolog (MVH) protein, a germ cell-specific RNA helicase, we have cloned genes that fail to be expressed in homozygous Mvh-knockout testes. One of the cDNA clones obtained by selective PCR subtraction cloning matched two cDNA sequences in the mouse testis cDNA database, and was highly homologous to Aven, a human gene regulating cell death. Sequences of the two mouse testis cDNAs in the database revealed that they share an 800 bp sequence but have quite different 5'-regions; the latter are derived from different exons by alternative use of transcription initiation sites. The shorter (mAven-S) and longer (mAven-L) transcripts encode 216 and 342 amino acids, respectively, and both isoforms are predominantly expressed in adult testes. However, mAven-L is not completely testis-specific and is expressed in testes during all developmental stages, whereas the expression of mAven-S is restricted to the testis and is barely detectable in Mvh-homozygous testes. In situ hybridization showed that mAven-S is expressed specifically in post-meiotic cells, mainly in round spermatids.
为了研究小鼠Vasa同源蛋白(MVH)的功能,一种生殖细胞特异性RNA解旋酶,我们克隆了在纯合Mvh基因敲除睾丸中未能表达的基因。通过选择性PCR消减克隆获得的一个cDNA克隆与小鼠睾丸cDNA数据库中的两个cDNA序列匹配,并且与Aven高度同源,Aven是一个调节细胞死亡的人类基因。数据库中两个小鼠睾丸cDNA的序列显示它们共享一个800 bp的序列,但5'区域有很大不同;后者通过转录起始位点的交替使用来自不同的外显子。较短的(mAven-S)和较长的(mAven-L)转录本分别编码216和342个氨基酸,并且两种异构体主要在成年睾丸中表达。然而,mAven-L并非完全睾丸特异性,在所有发育阶段的睾丸中均有表达,而mAven-S的表达仅限于睾丸,在Mvh纯合睾丸中几乎检测不到。原位杂交显示mAven-S特异性地在减数分裂后细胞中表达,主要在圆形精子细胞中。