Villegas Jaime, Araya Pamela, Bustos-Obregon Eduardo, Burzio Luis O
Bios Chile Ingeniería Genética SA, Millenium Institute for Fundamental and Applied Biology and Fundación Ciencia Para La Vida, Avenida Marathon 1943, Santiago, Chile.
Mol Hum Reprod. 2002 Nov;8(11):977-83. doi: 10.1093/molehr/8.11.977.
Amplification of RNA from human sperm heads yielded a fragment of 435 bp that shares 100% identity with a central region of the 16S mitochondrial rRNA. The nuclear localization in the sperm of the mitochondrial RNA was confirmed by in-situ hybridization. These results, together with the localization of the 16S mitochondrial rRNA in mouse sperm, are the first demonstration that the organelle transcript is a normal component of the mammalian gamete. The possibility that the nuclear mitochondrial RNA arises from nuclear transcription of a mitochondrial pseudogene was ruled out. To determine when during spermatogenesis the mitochondrial RNA is localized in the nucleus, in-situ hybridization of mouse and human testis was carried out. The nuclei of spermatogonia, spermatocytes and round and elongated spermatids were all positively stained. In human spermatocytes, the nuclear staining pattern was fibrillar, suggesting an association of the mitochondrial transcript with the meiotic chromosomes. These results indicate that early in spermatogenesis and before the onset of meiosis, the 16S mitochondrial rRNA is localized in the nucleus of spermatogenic cells, suggesting a process of translocation of the transcript from the mitochondria.
从人类精子头部提取的RNA经扩增后产生了一个435 bp的片段,该片段与16S线粒体rRNA的一个中心区域具有100%的同一性。通过原位杂交证实了线粒体RNA在精子中的核定位。这些结果,连同16S线粒体rRNA在小鼠精子中的定位,首次证明了细胞器转录本是哺乳动物配子的正常组成部分。线粒体RNA来源于线粒体假基因的核转录的可能性被排除。为了确定线粒体RNA在精子发生过程中的何时定位于细胞核,对小鼠和人类睾丸进行了原位杂交。精原细胞、精母细胞以及圆形和伸长的精子细胞的细胞核均呈阳性染色。在人类精母细胞中,核染色模式呈纤维状,表明线粒体转录本与减数分裂染色体有关。这些结果表明,在精子发生早期且在减数分裂开始之前,16S线粒体rRNA定位于生精细胞的细胞核中,提示该转录本从线粒体发生了易位过程。