Kleene K C, Distel R J, Hecht N B
Dev Biol. 1983 Aug;98(2):455-64. doi: 10.1016/0012-1606(83)90375-5.
We have isolated several cDNA clones encoding cytoplasmic poly(A)+ RNAs which are enriched in postmeiotic (haploid) spermatogenic cells in the mouse. Seventeen of 750 clones from a testis cDNA library hybridized more strongly to 32P-labeled cDNA copied from cytoplasmic poly(A) RNA of round spermatids than pachytene spermatocytes. Northern gel blots demonstrated that these 17 plasmids hybridized to RNA(s) approximately 0.5 kb (1 clone), 0.7 kb (13 clones), 0.8 kb (1 clone), and 0.9 kb (2 clones). Four plasmids hybridizing to RNAs 0.7 and 0.9 kb were further characterized by Northern blots. The levels of hybridization were about 10-fold greater with RNA from round spermatids, elongating spermatids and residual bodies than from pachytene spermatocytes from adult testis. These plasmids did not hybridize with cytoplasmic poly(A)+ RNA from sexually immature testis, adult liver, or brain, larger precursors in adult testis nuclear RNA, total RNA from cultured Sertoli cells, poly(A)- RNA from adult testis or the mouse mitochondrial genome. These results demonstrate that certain poly(A)+ RNAs are abundant in haploid cells but barely or not detectable in meiotic cells suggesting the accumulation of these RNAs in round spermatids requires transcription in haploid cells.
我们已经分离出了几个编码细胞质聚腺苷酸(poly(A))+RNA的cDNA克隆,这些RNA在小鼠减数分裂后的(单倍体)生精细胞中富集。来自睾丸cDNA文库的750个克隆中,有17个与从圆形精子细胞的细胞质聚腺苷酸RNA复制而来的32P标记的cDNA杂交,其强度高于粗线期精母细胞。Northern凝胶印迹显示,这17个质粒与大约0.5kb(1个克隆)、0.7kb(13个克隆)、0.8kb(1个克隆)和0.9kb(2个克隆)的RNA杂交。通过Northern印迹对与0.7kb和0.9kb RNA杂交的4个质粒进行了进一步表征。与成年睾丸粗线期精母细胞相比,来自圆形精子细胞、伸长精子细胞和残余体的RNA的杂交水平大约高10倍。这些质粒不与性未成熟睾丸、成年肝脏或大脑的细胞质聚腺苷酸+RNA、成年睾丸核RNA中的较大前体、培养的支持细胞的总RNA、成年睾丸的聚腺苷酸-RNA或小鼠线粒体基因组杂交。这些结果表明,某些聚腺苷酸+RNA在单倍体细胞中丰富,但在减数分裂细胞中几乎检测不到或无法检测到,这表明这些RNA在圆形精子细胞中的积累需要在单倍体细胞中进行转录。