(4)遺伝子発現プロファイリングによる卵の加齢機構の探索(<特集>第60回学術講演会要旨シンポジウム2「卵の発育・成熟・老化機構の解明と臨床応用」)
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Mammalian ooplasm supports preimplantation development, and reprograms an introduced nucleus transferred from a somatic cell to confer pluripotency in a cloning experiment. However, underlying molecular mechanisms of the oocyte competence remain unknown. Recent advent in microarray technologies has allowed us gene expression profiling of such tiny specimens as oocytes and preimplantation embryos. First of all, gene expression profiling of oocytes and preimplantation embryos revealed the distinctive patterns of maternal RNA degradation in addition to two major transient waves of de novo transcription, zygotic genome activation (ZGA) and mid-preimplantation gene activation (MGA). It has been thought that more than 90% of oocyte-stored RNAs are degraded by the 2-cell stage. Although our microarray analysis indeed confirmed the massive maternal RNA degradation pattern, we observed additional patterns of maternal RNA degradation showing significant reduction from the 4-cell to 8-cell stage. To elucidate molecular mechanisms maintaining oocyte competence and quality, we focused on global gene expression changes in age-associated loss of oocyte quality. we compared the expression profiles of metaphase II oocytes collected from 5-6 week old mice with those collected from 42-45 week old mice using the NIA 22K 60-mer oligo microarray. Among〜11,000 genes whose transcripts were detected in oocytes, about 5% (530) showed statistically significant expression changes, excluding the possibility of global decline in transcript abundance. Consistent with the generally accepted view of aging, the differentially expressed genes included ones involved in mitochondrial function and oxidative stress. However, the expression of other genes involved in chromatin structure, DNA methylation, genome stability, and RNA helicases were also altered, suggesting the existence of additional mechanisms for aging. Among the transcripts decreased with aging, we identified and characterized a group of new oocyte-specific genes, members of the human NACHT, leucine rich repeat and PYD containing (NALP) gene family. These results have implications for aging research as well as for clinical ooplasmic donation to rejuvenate aging oocytes.
- 2008-10-01
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