subcellular distribution of mitochondrial ribosomal rna in the mouse oocyte and zygote亚细胞分布的线粒体核糖体rna老鼠卵子和受精卵.pdfVIP

Subcellular Distribution of Mitochondrial Ribosomal RNA in the Mouse Oocyte and Zygote 1 2 Youichirou Ninomiya *, Shizuko Ichinose 1 Mammalian Development Laboratory, Department of Zoology, University of Oxford, Oxford, United Kingdom, 2 Instrumental Analysis Research Centre, Tokyo Medical and Dental University, Bunkyo, Tokyo, Japan Mitochondrial ribosomal RNAs (mtrRNAs) have been reported to translocate extra-mitochondrially and localize to the germ cell determinant of oocytes and zygotes in some metazoa except mammals. To address whether the mtrRNAs also localize in the mammals, expression and distribution of mitochondrion-encoded RNAs in the mouse oocytes and zygotes was examined by whole-mount in situ hybridization (ISH). Both 12S and 16S rRNAs were predominantly distributed in the animal hemisphere of the mature oocyte. This distribution pattern was rearranged toward the second polar body in zygotes after fertilization. The amount of mtrRNAs decreased around first cleavage, remained low during second cleavage and increased after third cleavage. Staining intensity of the 12S rRNA was weaker than that of the 16S rRNA throughout the examined stages. Similar distribution dynamics of the 16S rRNA was observed in strontium-activated haploid parthenotes, suggesting the distribution rearrangement does not require a component from sperm. The distribution of 16S rRNAs did not coincide with that of mitochondrion-specific heat shock protein 70, suggesting that the mtrRNA is translocated from mitochondria. The ISH- scanning electron microscopy confirms the extra-mitochondrial mtrRNA in the mouse oocyte. Chloramphenicol (CP) treatment of late pronuclear stage zygotes perturbed first cleavage as judged by the greater than normal disparity in size of blastomeres of 2-cell conceptuses. T